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AxiChrom™ 300-1000 columns - GE Healthcare Life Sciences

1. 164 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 4 System shutdown procedure WARNING Ensure that the power cable is disconnected or that the power is shut off at the outlet whenever maintenance work is performed in the Master to avoid the risk of electrocution WARNING A disconnecting system power switch must be installed such as a branch circuit protection and disconnect switch installed at your facility The system power switch should be installed at the wall to cut the power to AxiChrom Master Always use both the Master power switch located on the back on the Master unit and the system power switch for safe disconnection of AxiChrom Master Note When disconnecting the pneumatic pressure leading to AxiChrom Master a pop can occur Avoid this by first shutting off the pressure leading to the Master Procedure 1 Press the POWER OFF button in the MAIN MENU The dialog Saving data with progress bars is displayed briefly as in the following image Writing column data to CF card AxiChrom 300 1000 columns User Manual 28 9562 89 AD 165 7 After the packing 7 4 System shutdown procedure 2 When the following dialog is displayed use the power switch on the back of the Master to turn off the power IT IS NOW SAFE TO CAUTION If the power switch is turned off before the Saving data dialog has finished the saved data for the columns may become corrupted and
2. 145 Screw M10 X 25 146 Hex Head Screw ISO 4017 M12x45 A4 70 148 Hex Head Screw ISO 4017 M16x40 A4 70 149 Hex Head Screw ISO 4017 M16x45 A4 70 150 Hex Head Screw M12x45 151 Hex Head Screw M12x45 152 Hex Head Screw M12x45 154 Parallel Pin ISO 2338 12m6x60 A2 155 Foot M30 157 Hex Nut ISO 4032 M30 A4 70 159 Domed Cap Nut M10 160 Hex Head Screw ISO 4017 M6x16 A4 70 161 Washer ISO 7089 6 4x12x1 6 A4 163 Washer ISO 7089 13x24x2 5 A4 165 Washer 21x37 169 Washer Nylon 13x24x2 5 170 Washer Nylon 10 5x21x2 5 171 Motor 172 Hex Head Screw ISO 4017 M24x60 A4 70 173 M6S M20x200 A4 70 174 Hex Head Bolt ISO 4014 M16x80 A4 70 175 Hex Socket Head Cap Screw ISO 4762 M10x30 A4 70 178 Hex Socket Head Cap Screw ISO 4762 M8x30 A4 70 181 Hex Nut ISO 4032 M16 A4 70 182 Hex Nut ISO 4032 M8 A4 70 193 Pneumatic Connection Plug 194 Pneumatic Connection Plug 168 AxiChrom 300 1000 columns User Manual 28 9562 89 AD A Parts list and diagrams A 1 Part numbers and names 196 O ring 54x3 197 O ring 6 02x2 62 198 O ring 26 64x2 62 201 Column tube 202 Tie rod 203 Flange bottom 204 Flange top 211 O ring 673 1x9 525 213 Hex Head Screw M20x60 221 Hinge lower asm 231 Hex head screw ISO 4017 M16x45 A4 70 SSI column tube 232 Hex head screw ISO 4017 M16x50 A4 70 SS column tube 301 Bed support bottom 600 10 or 20 um 302 Bed support adapter 600 1
3. Column Unpacking Chrom 300 and ue L Cmn Aiheen 100 300 g Coan matens Acne vi Show medan type a Medem Cato DEAE E AxiChrom Option Description Column Select AxiChrom lt xxx gt lt yyy gt xxx column diameter in millimeter yyy max bed height for short and long column tube 300 or 500 mm respectively The column size can be found on a label located on the lower front part of the frame of the column Column material Select the material of the column tube Show medium type Select the appropriate medium type Medium Select the appropriate medium AxiChrom 300 1000 columns User Manual 28 9562 89 AD 147 7 After the packing 7 3 Unpacking the column 7 3 2 Creating a UNICORN method 3 Click on Next to display the Unpacking Parameters Method Wizard page Select the appropriate variables Description Measured bed height Enter measured value Inlet for unpacking liquid Select the appropriate inlet Capped outlet Select the appropriate outlet blocked with a blind cap 148 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 3 Unpacking the column 7 3 2 Creating a UNICORN method 4 Click on Next to display the Unpacking Summary Method Wizard page Hulliud Wiurl lub syleneantabiersss wuuhiunrcuntiol x Waste outlet can be changed in the variable pagel Outset Indole and estemated buller emmple consumption l
4. Note The tube installed on the Top mobile phase at delivery is only intended for storage solution conditioning and should be discarded once the column is in stalled The maximum operating pressure of the tube is 1 bar 3 2 Placing the column and Master Placing the column NOTICE Relieve the pressure from the column legs with a pallet jack or a lever while adjusting the legs or else the screw threads may get damaged NOTICE Make sure that the column stands exactly level after it has been moved If the column is not level this can impact column operations such as packing and the resulting bed efficiency 1 Use a pallet jack or the lever used for assembling wheel kits to relieve the pressure from the column legs while adjusting them 2 Carefully check for wobble Make sure there is even load on all 4 feet 42 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 2 Placing the column and Master 3 A spirit level placed diagonally on top of the column will show when the column stands horizontal Ensure that the column is level in all directions by measuring with the spirit level on different places of the column 4 When the column feet have been adjusted tighten the lock nuts on the feet using a tightening torque of 70 Nm Placing Master Place AxiChrom Master outside the AxiChrom column safety zone A Description of the safety zones AxiChrom 300 1000 col
5. Maintenance Low pressure 0 5 bar High pressure 4 bar 2 4 Materials Background information The materials used to manufacture AxiChrom columns have been chosen for their bio logical and chemical compatibility with the solvents used during operation and cleaning in place CIP procedures The columns have also been designed to comply with the varying hygienic requirements at the different stages of development and production Polymer materials in AxiChrom columns in contact with process liquids have been selected for their biological compatibility according to the United States Pharmacopeia USP Bi ological Reactivity Tests In vivo and conform to USP class VI requirements compliance with Code of Federal Regulations CFR Food and Drug Administration Title 21 Part 177 and being animal free or complies with the conditions in the CPMP Note for Guidance EMEA 410 01 Rev 2 Column tubes for AxiChrom 300 1000 columns are available in acrylic plastic or stainless steel Use and maintenance of AxiChrom columns with stainless steel column tubes differs from that for acrylic plastic column tubes in two major respects e Stainless steel column tubes are not transparent so the bed cannot be observed directly This affects packing and unpacking procedures e Stainless steel column tubes are manufactured in a single piece with no removable tie rods top or bottom flanges or corresponding O rings This does not affect normal
6. AxiChrom 300 1000 columns User Manual 28 9562 89 AD 117 5 Packing the column 5 3 Packing using a UNICORN method 118 In the Variables page enter the Measured slurry concentration see the figure below and the information in Section 4 3 Determining the slurry concentration on page 91 Vananes Block Te Variable Value Range Main Theoretical_CV I 157 080 0 100 999999 000 E Alarm_Pressure Pressure_Limit bar 4 00 0 00 7 00 Pressure _Level Warning bar 4 00 0 00 7 00 Pressure_Limit_2 bar 4 00 0 00 7 00 Pressure_Level_Warning_2 bar 4 00 0 00 7 00 Hose_Properties Hose_InDiameter mm 22 3 0 0 100 0 Hose_Length m 2 00 0 00 10 00 Packing_Settings_1 Accepted_Slury_Concentration Slurry_Concentration 70 80 Inlet Rinsing Inlet Rinsing J SystemWash_Mobile Inlet_Mobile InletAl Volume_System Wash_ Mobile Molume_System Wash_Mobile 1 12 00 0 00 999999 00 Colurnn_E quilibration Volume_Column_Equilibration CV 1 00 0 00 999999 00 Flow_Rate_Equilibration Flow_Rate_Equilibration hour 785 4 0 0 2000 0 Inlet_SampleLoad Inlet_SampleLoad inlets Volume_Systemwash_Sample Volume _Systemwash Sample i 1200 0 00 999993 00 Upflow_SampleLoad_Peak Flow_Rate_SampleLoad_Upflow I hour 235 6 0 0 2000 0 iv Show details Show unused variables 7 Display tooltip for extended variable cells Help Click Next
7. For rupture discs used on AxiChrom 800 1000 the T junction 28 9381 70 can be used Gemii valves Article name Code no Gem valve ID 7 75 2xTC25 28 9382 05 Gem valve ID 9 4 2xTC25 28 9382 06 Gemt valve ID 15 75 2xTC25 28 9382 07 Gemt valve ID 22 1 2xTC50 28 9382 08 Gemt valve ID 34 8 2xTC50 28 9382 09 Valve holder kit Article name Code no Valve holder kit 29 0137 10 Media stirrer Article name Code no Media stirrer 80 mm 28 9191 03 Media stirrer 150 mm 28 9191 04 190 AxiChrom 300 1000 columns User Manual 28 9562 89 AD C Accessories and additional literature Additional literature Article name Code no Sanitization and endotoxin clearance in AxiChrom columns 28 9290 42 Application note Column efficiency testing 28 9372 07 Application note AxiChrom 300 1000 columns User Manual 28 9562 89 AD 191 D Crate weights and dimensions Appendix D Crate weights and dimen SIONS Typical weights and dimensions for the different crate sizes are listed in the table below Column Column bed Crate Crate Crate Total weight diameter heights height length width crate col mm mm mm mm mm umn kg 300 100 to 300 1920 970 1495 160 574 580 300 100 to 500 2180 970 1495 165 599 605 400 100 to 300 1920 970 1495 160 611 620 400 100 to 500 2180 970 1495 165 636 645 450 100 to 300 1920 1090 1480 180 879
8. e PRIMING guides the user through safe priming of the column e INTELLIGENT PACKING with pre programmed packing meth ods it guides the user through packing of the column to achieve optimal compression and bed efficiency e UNPACKING guides the user through safe unpacking of media from the column e MANUAL CONTROL enables the user to control the adapter movement and open or close the Media valve e MAINTENANCE guides the user to set the column into Mainte nance mode from where safe service of the column can be performed See AxiChrom Operating Instructions e POWER OFF from this fail safe mode it is safe to turn off Axi Chrom Master without loss of information 76 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 7 Using the Master 3 7 2 User interface Button General interface functions HELP Opens contextual help texts information for the current dialog From the HELP dialogs it is possible to open a Flow converter for conversion of linear to volumetric flow The HELP button is normally grey when it is active In the column list the HELP button turns blue if the marked column has corrupt column information BACK Opens the previous dialog The BACK button is blue when it is active CONTINUE Opens the next dialog in the workflow This button is only active blue when all mandatory information has been entered in the active dialog In some wizards it is possible
9. Ground See Grounding the column on page 65 Power supply cable 3 on the Master Power supply connector 380 400 VAC 50 60 Hz with protective ground The AxiChrom Master is delivered with CE or UL approved cables AxiChrom 300 1000 columns User Manual 28 9562 89 AD 61 3 Getting started 3 4 Connections Recommended mobile phase tubing inner diameters 62 Note Table 3 1 Tubing inner diameters recommended for top and bottom mobile phase connections for different column inner diameters All dimensions are given in millimeters A dash means that the combination is not compatible with Intelligent Packing TC25 connectors do not have the same inner diameters as the tubing See Tubings on page 187 for details AKTAprocess 300 400 450 600 800 1000 dimension 6 mm PP 5 9 4 9 4 9 4 6 4 6 4 6 4 3 8 SS 9 4 9 4 9 4 6 4 6 4 6 4 9 4 10 mm PP 9 4 9 4 9 4 12 7 12 7 12 7 12 7 19 1 6 4 6 4 6 4 9 4 1 2 SS 9 4 9 4 9 4 12 7 12 7 12 7 12 7 19 1 9 4 254 25 4 1 PP and SS 12 7 34 7 34 7 19 1 Use narrower tubing for packing gel filtration or polishing media Use the widest tubing compatible with the system for high flow media such as Capto AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 4 Connections Power requirements and connections The general requirement
10. PM Offset 03724 The Calibration page will synchronize the Master unit with the motor The column may not function with the Master unit if the motor is not calibrated A calibration is therefore always performed when a new column is selected Calibration will start automatically and will normally take less than one minute to com plete When the calibration is completed the CONTINUE and BACK buttons will turn blue e Press CONTINUE to proceed to the position verification page e Press BACK to return to the column selection page Note If several columns are to be calibrated in a row the Master unit must be shut down and restarted between calibrations AxiChrom 300 1000 columns User Manual 28 9562 89 AD 81 3 Getting started 3 7 Using the Master 3 7 3 Column and Master setup Verify the adapter position 82 The actual adapter position on the column has to be entered in the AxiChrom Master Follow the procedure below to correctly read and enter the correct adapter position in the dialog Verify adapter position Verify that the last known adapter position agrees with the position measured on the scale on the column If they do Not agree enter tho correct position as measured on tho Adapter position 9 0 cm scale CAUTION It is very important that the correct position of the adapter re enturod Last known adapter position 9 0 cm Pressure 0 28 bar Adaptor stopped Modia valve closed 1 Verify the actual dist
11. 2 Enter a low adapter speed 30 cm h is recommended 5 Enter the target height higher than the current height but within the earlier applied compression range 4 The adapter will move to the target height 5 When the adapter stops stop the liquid flow through the column 6 Close the inlet 7 Evaluate the column performance see Chapter 6 Evaluating column performance on page 141 Method 3 Another post packing modification is flow conditioning suitable if the bed shows signs of over compression at one end such as markedly different HETP and A results in the upflow and downflow directions there are cracks in the bed HETP test results indicate a bed with insufficient efficiency Flow conditioning can sometimes correct this there are shoulders on the HETP peak which indicates poor packing AxiChrom 300 1000 columns User Manual 28 9562 89 AD 139 5 Packing the column 5 5 Creating custom packing methods in the Master 5 5 2 Post packing modifications 140 For more information about HETP calculation see Chapter 6 Evaluating column perfor mance on page 141 To flow condition the bed it is important to have a pump that can supply a flow high enough to create a slight head space of clear liquid above or below the bed If this cannot be achieved run the pump media and column at the highest flow they can withstand Procedure 1 Start the pump in one direction through the column and slowly increase the flow to at least the opera
12. phase connection is detected automatically AxiChrom 300 1000 columns User Manual 28 9562 89 AD 63 3 Getting started 3 4 Connections WARNING Ensure that the EMERGENCY STOP button can always be reached while working with the column WARNING Electrical installations must be performed by authorized personnel only WARNING Only authorized personnel are allowed to open the cabinet of Axi Chrom Master There are no user serviceable parts inside the cabinet WARNING Always connect AxiChrom Master to a power supply with protective grounding WARNING Connection to wrong power supply voltage may cause injury to personnel and damage to the system WARNING The power cables must be replaced only by cables of the same type or equivalent fitted with the same type of connectors Power cables and connectors must be replaced or repaired by properly trained personnel authorized by GE Healthcare WARNING The power connection must be easily accessible to enable the user to disconnect the power in case of an emergency 64 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 4 Connections NOTICE Connect the communication cable and power cable for the adapter control motor on the column to AxiChrom Master before AxiChrom Master is connected to power Note This is a class A product input power gt 1 kW intended for profession
13. 1000 columns User Manual 28 9562 89 AD 4 4 Preparations before packing the column 4 2 Preparation of the slurry Prepare a sufficient amount of slurry in the slurry tank to be able to pack the desired bed height The required amount of slurry is also automatically calculated in the AxiChrom Master Intelligent Packing wizard and displayed in the summary page before the packing is started Use CF for gravity settled media Slurry volume target bed height x PF x column cross section slurry concen tration Prime the media valve Make sure that all valves in the slurry tank are closed and that the gel can not flow into any tubing where it will sediment Use a media stirrer to make ho mogenous slurry in the tank before packing Tip For Manual operation e Max fill height for short column tubes bed height 100 to 300 570mm e Max fill height for long column tubes bed height 100 to 500 830 mm 4 3 Determining the slurry concentration Choice of methods Two methods are presented here for determining the slurry concentration Forced settling Gravity settling Forced settling To achieve the most accurate result a general determination method for slurry concen tration has been developed The method utilizes a laboratory column preferably XK16 or HiScale 16 and a small pump that can deliver 100 cm h for the specific column Preparations 1 2 Mount an end piece adapter with a filter to the bot
14. AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1 Introduction 1 2 Scope of this User Manual Chapter 4 Preparations required before packing the column including slurry preparation and priming the column Chapter 5 Overview of the Intelligent Packing procedure and instructions on how to pack the column using UNICORN or AxiChrom Master interface Chapter 6 Guidance how to evaluate the performance of the column and perform HETP calculations Chapter 7 Instructions for performing CIP Sanitization in place SIP un packing the column using UNICORN or Master interface and shutting down the column and Master Appendices Spare parts information and exploded diagrams Tubing kits and connections Packing buffers and recommended packing factors Column and crate weights and dimensions Decontamination report Associated documentation The following documentation is provided with the AxiChrom 300 1000 columns Manual name Document num Description ber AxiChrom 300 1000 28 9562 89 This manual columns User Manual AxiChrom 300 1000 28 9562 90 Safety information and instruc columns Operating Instruc tions for installation operation and tions maintenance Includes a CD with translations into local languages AxiChrom columns Site Configured for Guidelines and requirements re Preparation Guide individual garding the installation site and columns how to place the column AxiChrom 300 1000 28 9562 91 Instructions o
15. AxiChrom 300 1000 columns User Manual 28 9562 89 AD 149 7 After the packing 7 3 Unpacking the column 7 3 2 Creating a UNICORN method e Regular unpacking method should be used for media with a medium particle size d from approximately 50 ym and above The method consists of a bed collapse part and a slurry push out part e Advanced mode should be used for media with d less than 50 um The method consists of several bed collapse parts and several slurry push out parts The sequence for advanced mode is repeated two or three times depending on bed height A bed height below 25 cm is unpacked with two slurry push outs and a bed height above 25 cm is unpacked with three slurry push outs Regular unpacking is the fastest method If the unpacking behavior is unknown try the regular method 7 3 3 Unpacking acrylic column tubes 150 Unpacking acrylic column tubes using UNICORN method This procedure assumes that the column system and tubing between the system and the column are primed The unpacking sequence is the same as for the AxiChrom Master in Unpacking acrylic column tubes using Master wizard on page 151 from Step 7 Start the pump upflow When UNICORN controls the unpacking all valves and flow path are automatically changed The only operation controlled by the operator is to open the Slurry valve and make some confirmations The recommended unpacking solution is water If the media is to be stored unpack in appropriate storage
16. Getting started 3 7 Using the Master 3 7 3 Column and Master setup Perform a leakage test as described in the AxiChrom 300 1000 Operating Instructions when e the column is new e the column has been shipped e the o rings have been replaced e other parts that may cause leakage have been replaced e maintenance has been performed 3 8 Cleaning the column The cleaning should be performed before the bed is packed or when the column is new that is when the column is empty or filled with transportation storage solution Cleaning procedure Note 1 Parts made from non plastic materials may be autoclaved For more information about cleaning methods see the User Manual Clean the outside of the acrylic tube with for example water and or 20 v v ethanol to remove any surface residues from the packaging NOTICE Do not use ethanol stronger than 20 v v for cleaning the acrylic tube since this may damage the column tube Steel surfaces can be cleaned with a wetted cloth and 70 v v isopropanol or 70 v v ethanol Ensure that the adapter seals and o rings are clean from particles and waste since these may damage the adapter and tube when the adapter is moving Perform a priming according to the instructions in the PRIMING wizard in the AxiChrom Master MAIN MENU This procedure will clean the column and its tubes and remove any air from the column Ensure that the cleaning solution is comple
17. Introduction 1 5 Intended use of the columns Approved operator The use of interactive guides in AxiChrom Master and in the Intelligent Packing wizard allows consistent performance with reproducible quality To use the column safely and correctly the operator must have read understood and be fully acquainted with AxiChrom 300 1000 columns User Manual and AxiChrom 300 1000 columns Operating Instructions Improper operation of the column may result in property damage as well as personal injury 1 6 Safety precautions Introduction Before installing operating or maintaining the AxiChrom column you must be aware of the hazards described in this manual Follow the instructions provided to avoid personal injury or damage to the equipment The warnings and cautions in the user documentation shall in no way take precedence over more restrictive local regulations and policies For your personal safety it is important that you have a proper knowledge of the entire system that the column is part of Study any complementary safety instructions and use appropriate personal protection equipment for the specific application and operation environment Risk assessment AxiChrom columns have been designed and manufactured to provide a high level of personal safety However the residual risk is highly dependent on the application and environment in which the column is operated In order to determine the safe operation of the equipment a risk
18. Result The adapter automatically moves down to 30 cm at a velocity of 300 cm h expelling air and priming solution through the top bed support and the Top mobile phase At 30 cm the flow direction in the system is automatically changed so that the bottom bed support and bottom mobile phase are primed while the adapter moves down to the bottom position of 1 0 cm 6 The priming of the Media valve column and system is now complete 7 Exchange the liquid in between the scraper seals to 20 ethanol 4 4 5 Using Master wizard The Priming procedure described in the wizard guides the operator through a number of steps to prime the column with a standalone pump Follow the instructions in the PRIMING wizard found in the MAIN MENU in AxiChrom Master For further explanation of the buttons used in the wizard refer to Buttons and explana tions on page 76 For tubing connections see Manual configuration on page 69 Note Before the column is primed the column must also be calibrated page 75 and cleaned page 85 also when priming with UNICORN WARNING Ensure that nobody is present within the safety zone when the adapter is moving The operator in charge is responsible for keeping the safety zone free from unauthorized personnel and unnecessary equipment CAUTION Always open and close external valves slowly Sudden changes of pressure in combination with air in the column might cause damage to the column For fu
19. Step 9 create back pressure Priming progress reate back pressure 8 15 een Create back pressure Adapter position 61 8 cm Tighten the valve on the top mobile phase until the pressure is J approximately 0 3 bar Press CONFIRM to continue Pressure 0 38 bar Adapter stopped Media valve closed Create back pressure by tightening the membrane valve on the Top mobile phase until the pressure is approximately the value stated in the dialog Check the box when completed Press CONFIRM to proceed AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 4 Priming the column 4 4 5 Using Master wizard Step 10 wait for the adapter to move down past the priming groove Priming progress Moving the adapter down 9 15 wam 238 romaning Adapter commanded f peau S75 on TETT Moving the adapter down The adapter moves down at 40 cmh passing the Priming groove expelling the air under the adapter through the seal flush valves Pressure 0 78 bar Adapter moving Media valve closed The adapter now moves down at 40 cm h passing the priming groove expelling the air under the adapter through the Seal flush valves Press PAUSE to stop the movement temporarily or to terminate the priming wizard Step 11 release the pressure Priming progress Release pressure 10 15 enue Release the pressure Adapter position 57 8 cm Close the seal flush valves Stop the pump i Slovty ope
20. XL DEAE Sepharose Fast Flow Other iy piae an _ aan Select an available medium for the selected chromatography technique that was chosen in the previous dialog A scrollbar will appear on the right side of the list if it is too long to fit on the screen All lists include a selection called Other which is to be used for media other than GE Healthcare BioProcess media Press the CONTINUE button to proceed 122 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 4 Packing using the Master Wizard Step 4 Enter the hose dimensions Enter hose dimensions 4 15 enna Capto Q Slurry hose inner diameter 22 1 mm E a Slurry hose length 2 00 m en Cell atl nama uai e Enter the slurry tube inner diameter in millimeters by pressing Set if the current value is wrong e Enter the slurry tube length in meters by pressing Set if the current value is wrong Note If the wrong tubing parameter is set the bed height can be affected Press the CONTINUE button to proceed Step 5 Enter the target bed height and the slurry concentration Enter packing settings 5 15 waw Capto Q Enter slurry concentration 65 Ce al Enter target bed height 10 0 cm E Sem Inteligent Packing bed height limits Enter max imit 11 0 cm Ce 1 eaan Enter min limit 9 0 cm Peal a AxiChrom 300 1000 columns User Manual 28 9562 89 AD 123 5 Packing the column 5 4 Packing using the Master Wizard e Press Set if the current value
21. adapter moves upwards under upflow 2 The pressure rises again abruptly when the adapter is in the top position and the flow is reversed 3 The pressure rises further as media settles on the bottom bed support 7 4 System shutdown procedure Background The AxiChrom Master should always be turned off disconnected and removed after packing unpacking maintenance procedures and before the column is used for sepa ration runs Note If UNICORN has been used to control the Master make sure that the component in UNICORN System setup Edit Component for Intelligent Packing Axi Chrom 300 1000 is unticked before disconnecting the Profibus cable as the following image shows AxiChrom 300 1000 columns User Manual 28 9562 89 AD 163 7 After the packing 7 4 System shutdown procedure Component xj Select components i Process1 v CIP amp IntelligentPacking AxiChrom 100 200 AxiChrom 300 Cancel Help Disconnecting KTAprocess from AxiChrom Master Follow the instructions on page 74 Shutdown procedure for AxiChrom Master Use this procedure to shut off and disconnect the Master safely WARNING Disconnect the power cable from the power outlet to AxiChrom Master before disconnecting the power cable between the Master and column There is still lethal voltage present in AxiChrom Master when it is shut off using the power switch on the Master
22. assessment must be made This risk assessment in combination with local regulations and policies will result in specific safety instructions for installation operation and maintenance use of proper personal protective equipment or other ar rangements that are needed to operate your process safely AxiChrom 300 1000 columns User Manual 28 9562 89 AD 13 1 Introduction 1 6 Safety precautions General precautions WARNING The customer must ensure that all installation maintenance operation and inspection is carried out by qualified personnel who are adequately trained understand and adhere to local regulations and the operating instructions and have a thorough knowledge of AxiChrom 300 1000 columns and the entire process WARNING AxiChrom columns must NOT be used for any other purposes than chromatographic separations They must not be used as e Storage tanks for chemicals etc e General pressure vessels e Fermentation vessels e Gas storage tanks The column must never contain air or gas under pressure Using flammable liquids WARNING AxiChrom columns are not designed to handle flammable fluids AxiChrom is not approved for work in a potentially explosive atmo sphere in areas classified as Zone 0 to Zone 2 according to IEC 60079 10 2002 14 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1 Introduction 1 6 Safety precautions Personal protection WARNING
23. bed support nuts Column Tightening torque 300 600 10 Nm 800 1000 15 Nm 88 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 Preparations before packing the column 4 1 Packing Factor and Compression Factor Introduction During a standard AxiChrom packing procedure the media is first settled under a low flow induced by the adapter moving downwards This is what we refer to as consolidation The Top mobile phase is closed and the Bottom mobile phase is open during packing and the excess liquid is expelled through the Bottom bed support on which the bed will start to form When all media has settled and formed a bed at the bottom of the column the bed is consolidated still under influence of a low flow until the adapter hits the bed Adapter movement then continues compressing the media until the required packed bed height is reached based on the packing factor used The Packing Factor PF is cal culated as follows packing factor PF consolidated bed height packed bed height The Packing Factor takes the settling speed of media into account The Compression Factor CF is defined as the ratio when no flow through the bed is applied compression factor CF gravity settled bed height packed bed height There is a difference between Packing Factor and Compression Factor The definition of Packing Factor involves flow through the bed which in some cases
24. cm h is used for the whole process Note Before the sanitization method is started check that the media withstands 1 0 M NaOH Regular cleaning 144 Regularly wipe the outside of the column using a cloth moist with an antibacterial agent This reduces the risk of microbial growth AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 2 Sanitization in place General method for sanitization in place il It is recommended to rinse the column and Media valve with purified water 2 column volumes upflow at a flow velocity of 60 cm h Clean the Media valve by filling it through the Rinse inlet and out through the Slurry inlet with 1 M NaOH Let stand for 4 hours before neutralization Mean while continue with the next step mn ush the column upflow with 2 column volumes of 1 0 M NaOH at a velocity of 60 cm h mal ush the column downflow with 2 column volumes of 1 0 M NaOH at a velocity of 60 cm h D ecirculate 5 column volumes of 1 0 M NaOH upflow for 4 hours at a velocity f 60 cm h Wash the column and Media valve with buffer or water of desired quality to achieve a neutral pH Re equilibrate the column with storage or starting buffer An application note on sanitization is available from GE Healthcare 7 3 Unpacking the column Section contents This section first gives an overview of the unpacking procedure Later in this section there is a more detailed stepwi
25. in the shape of the peak is usually the first indication of bed deterioration The asymmetry factor A describes the deviation from an ideal Gaussian peak shape and is calculated from the peak width at 10 of peak height A b a The following test chromatogram shows the parameters used to calculate peak broad ening and peak symmetry Concentration measured by Absorbance or Conductivity AxiChrom 300 1000 columns User Manual 28 9562 89 AD 143 7 After the packing 7 After the packing 7 1 Cleaning in place CIP Description Efficient cleaning methods should be developed as an integral part of the complete process to maximize the working life of the medium CIP removes precipitated material strongly bound substances and other contaminants from the medium without the need to disassemble the column Regular CIP between batches helps assure the desired product quality and the expected working life of the medium and equipment For more information about which chemicals to use for CIP see the table on page 37 Note Ensure that the media and the column withstands the chemicals used for CIP 7 2 Sanitization in place Description Sanitization is the use of chemical agents to minimize microbial contamination A sanitization procedure is used when there is a need to ensure microbial reduction for example before changing a product or when there has been unwanted microbial con tamination Low fluid velocity 60
26. is connected to the Slurry inlet A hose from the pump system is connected to the Rinse Inlet The adapter is in the bottom position lt BACK CONTINUE gt The AxiChrom Master Intelligent Packing wizard starts with a welcome dialog where all necessary preparations before packing are listed You can step back and forth in the wizard until the START button has been pressed step 8 The preconditions for packing are The system pump is primed All tubing leading to the column are primed The column is primed A slurry tank is connected to the Slurry inlet A tube from the pump system is connected to the Rinse inlet The adapter is in its bottom position Press the CONTINUE button to proceed AxiChrom 300 1000 columns User Manual 28 9562 89 AD 121 5 Packing the column 5 4 Packing using the Master Wizard Step 2 Select the media type Select media type 2 15 e00vs00 Media type D Affinity Anion exchange a eee Cation exchange Metal affinity Hydrophobic interaction Size exclusion Sah ae wos Select a medium type for a certain chromatography technique using the arrow buttons The selection All types will list all available media regardless of their applications Press the CONTINUE button to proceed Step 3 Select the medium Select medium 3 15 1O00G00sara Medium Scanio Adhere Capto DEAE Capto Q meen Sst Q Sepharose Big Beads 2 z Q Sepharose Fast Flow Q Sepharose High Performance Q Sepharose
27. is to be Follow the instructions in Section 3 8 Cleaning the column used soon on page 85 The column is to be Unpack the column from the crate check it and clean it used at a later time according to the instructions below within 3 months from Note delivery If the column is not cleaned there is a substantial risk of mold and corrosion attacks Cleaning procedure 1 Remove the tri clamp blind cap from the Bottom mobile phase inlet outlet note that the column may be under slight pressure Connect a pump to the Bottom mobile phase inlet outlet e Ensure that the tube from the pump is primed that is completely filled with liquid before it is connected to the Bottom mobile phase inlet outlet e The pump must be switched off during installation so that the column does not get pressurized Remove the tri clamp blind cap from the tube at the Top mobile phase and connect a tube that leads to waste Flush the column with at least 6 column volumes calculated from an adapter position of 1 to 2 cm at delivery of 20 ethanol Prepare the column for storage according to AxiChrom Operating Instructions AxiChrom 300 1000 columns User Manual 28 9562 89 AD 41 3 Getting started 3 1 Unpacking the crates and cleaning a new column NOTICE If the column is filled and should be moved plug the Top mobile phase and Bottom mobile phase inlets with tri clamp blind caps diaphragms or other suitable valves
28. is wrong and enter the measured slurry concentration in percent If a slurry concentration above 70 is entered the default filling speed will be decreased to 200 cm h e Enter the target bed height in centimeters by pressing Set if the current value is wrong The value is set to one decimal place accuracy e The deviation from the desired target bed height is automatically suggested by In telligent Packing bed height limits The default range is 1 cm of the target bed height Press Set to adjust the values Note The more accurate the slurry concentration is the closer the bed height will be to the nominal target bed height Set a range for the bed height limit to allow some tolerance in slurry concentration Press the CONTINUE button to proceed Step 6 Confirm the packing settings Confirm the packing settings Use the recommended default values for Packing Factor Filling speed and Packing speed These have been verified for GE Bioprocess media nfirm packir attinas ana Capto Q The default settings have been verified to give well packed beds Target packing factor 1 10 Filling spoed 300 cm h Packing speed 60 em h CONTINUE e Press the CONTINUE button to use the default settings that have been verified to give well packed beds e Press Edit to change any of the values not recommended other than for reasons stated in Section 4 2 Preparation of the slurry on page 90 Then press Set in the next dialog to chang
29. leads to a more compressed bed and lower consolidated bed height while in other cases the gravity settled bed height is lower since the bed packs more densely if given more time to do so Mabselect SuRe for example has a significantly lower gravity settled height than its consolidated height and this has to be considered when calculating the amount of media to be packed in the column About default packing values The default values in the Master software are tested and verified parameters for packing in water 20 ethanol Packing Factor is allowed to vary around the optimal value 0 03 which has also been verified e For HIC media the default values are defined for packing in 10 to 20 ethanol in the AxiChrom 300 to 1000 columns AxiChrom 300 1000 columns User Manual 28 9562 89 AD 89 4 Preparations before packing the column 4 1 Packing Factor and Compression Factor e For some media packed in salt containing solution a higher Packing Factor has to be used This is due to different settling conditions in solutions other than water and ethanol e Note that all S and SP ligand media are shipped and stored in 0 2 M sodium acetate and any decanted solution will contain salt For detailed information about which Packing Factor to be used depending on packing buffer see Appendix B Packing buffer information on page 183 Bed support porosity 4 2 90 The bed support porosity should be less than or equal to 1 3 of the av
30. light grey Virtual keypad The virtual keypad is used to enter input for text and figure fields AxiChrom 300 1000 columns User Manual 28 9562 89 AD 77 3 Getting started 3 7 Using the Master 3 7 2 User interface Information in progress dialogs During packing unpacking and priming the progress can be monitored in progress di alogs They display the remaining time and a graphical progress bar the adapter position shown both graphically in the column drawing and in the adapter position box actual pressure in the column the state of the bottom Media valve the packing process steps shown in text and checked when completed new status messages The action buttons will change functions depending on the specific status of the packing process The status message may call for some action from the user for example to open the Bottom mobile phase valve or clean the valve and tube The action will then require a confirmation before the process is resumed 3 7 3 Column and Master setup Select column 78 Gon CONFIGURATION CONTINUE gt If the column to be used can be seen in the list select it with the arrow buttons If the column to be used can not be seen in the list press Add to add a column to the list No column will be listed when Master is delivered Press Edit to change the parameters of the chosen column or the column name Press Delete to remove the chosen column from the list AxiCh
31. operation or maintenance procedures but results in a spare parts and accessories list that differs from that of acrylic columns Parts list and materials Component Material In contact with process stream Adapter backing plate Stainless steel ASTM 316 or ASTM No 32205 Adapter seals and snap UHMWPE ultra high molecular weight Yes ring polyethylene AxiChrom 300 1000 columns User Manual 28 9562 89 AD 35 2 Specifications and characteristics 2 4 Materials Component Material In contact with process stream Bed support Stainless steel ASTM 316L and ASTM Yes 32205 or PE polyethylene or UHMWPE Column tube PMMA polymethyl methacrylate or Yes Stainless steel ASTM 316L Distributor PP polypropylene Yes Dynamic seals FFPM full fluorinated propylene Yes monomer or UHMWPE Media valve body PP Yes Static seals EPDM ethylene propylene diene Yes monomer Top mobile phase PP Yes Bottom backing plate Stainless steel ASTM 316 No Lid Stainless steel ASTM 316 No Stand Stainless steel ASTM 316 No 2 5 Chemical resistance AxiChrom columns are resistant to chemical agents used in protein recovery including buffer solutions for adsorption elution and washing and to solutions effective in cleaning sanitization and storage Table lists chemicals that may or may not be used with AxiChrom columns The concentrations listed are not normally exceeded during an
32. solution such as 20 ethanol or 10 mM NaOH Note One outlet should be capped during the unpacking to block the Top mobile phase outlet during the media push out step when the Media valve is open to the slurry tank WARNING Ensure that nobody is present within the safety zone when the adapter is moving The operator in charge is responsible for keeping the safety zone free from unauthorized personnel and unnecessary equipment Before running the method Connect the column using the automatic configuration for AktaProcess as described in Section 3 6 Column and system setup on page 68 Running the method Ensure that the liquid levels in the containers are lower than the system outlet This is to avoid siphoning flow AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes 1 Start the Unpacking method Result The bed expands and collapses To speed up the bed expansion sequence increase the adapter speed with Pump Increase_Adapter_Speed under Manual Instructions The adapter speed can be increased in increments of 30 cm h up to 240 cm h Note that the pump flow velocity should always be higher than the adapter speed If the bed collapses and comes to the bottom of the column earlier the com mand Next breakpoint in Manual Other in UNICORN can be used to speed up the process 2 When prompted to do so open the slurry tank Result Th
33. stainless steel bed support Some crate weights are estimates AxiChrom Master weights and dimensions Parameter Value Weight kg 73 Length mm 670 Width mm 590 Height mm 1090 194 AxiChrom 300 1000 columns User Manual 28 9562 89 AD F Decontamination report Appendix F Decontamination report Decontamination Report 1 Equipment that is returned for service or any other reason when personnel connected to GE Healthcare must handle the equipment at any location should be cleaned 2 This form must be used to log the decontamination of the equipment 3 A completed copy of this form should be faxed or sent by first class post to the person who will come in contact with the equipment to ensure that he she has the information before handling the equipment 4 Failure to complete the form or comply with the procedure will lead to costs for decontamination and delays in hand ling the equipment 5 Company Address Phone Fax number 6 Please complete the following sections 6 1 Equipment type 6 2 Serial number 6 3 Details of substances used on the inside outside of the equipment 63 1 Substance namels 6 3 2 Precautions to be taken in handling these substances 6 3 3 Action to be taken in the event of human contact 6 3 4 Cleaning fluid to be used if residue of chemicals is found during handling 6 4 Any further relevant information 7 If
34. steel ASTM 316 and ASTM316L EN 1 4401 1 4436 and EN 1 4404 1 4432 1 4435 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 31 2 Specifications and characteristics 2 3 Primary specifications 2 3 Primary specifications 2 3 1 Weights volumes and related dimensions Column 10 0 400 450 31010 310 0 sKolole Tube inner diameter A 300 400 450 600 800 1000 mm Footprint BxC mm 1110 x 1110 x 1110 x 1180 x 1470 x 1720 x 520 600 620 780 1080 1300 Safety zone mm 3200 x 3200 x 3200 x 3200 x 3500 x 3800 x 2600 2600 2700 2800 3100 3300 Column cross section cm2 707 1257 1590 2827 5027 7854 Max bed volume 21 35 36 63 48 80 85 141 151 251 236 393 short long tube liters Weight of empty column 420 440 460 480 710 760 835 900 2150 2240 2560 2680 short long column stainless steel bed support kg Weight of empty column 414 434 451 471 699 749 818 883 2122 2212 2517 2637 short long column plastic bed support kg 32 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 2 3 2 Heights in different operating states 2 Specifications and characteristics 2 3 Primary specifications 2 3 2 Heights in different operating states Note The letters in parentheses refer to the image on page 32 All dimensions are given in millimeters Column Tube 300 400 450 31010 800 KKOO Max height D Short 22
35. system after an emergency stop WARNING The column may still be pressurized when the process has been stopped using the EMERGENCY STOP button Ensure that the pressure is released before disconnecting any tubing The EMERGENCY STOP button will abort the process that is currently in progress in Axi Chrom Master for example Intelligent Packing The process has to be restarted from the beginning after the emergency has been resolved If the EMERGENCY STOP is pressed during the MAINTENANCE wizard or MANUAL CONTROL wizard the MAIN MENU is displayed When the assembly or disassembly process is launched again the software returns to the same state as when the button was pressed The EMERGENCY STOP button is released by turning the button counter clockwise If the column was being packed the column must be emptied using MANUAL CONTROL or UNPACKING in AxiChrom Master or the unpacking method in UNICORN before the Intelligent Packing wizard can be started again AxiChrom 300 1000 columns User Manual 28 9562 89 AD 21 1 Introduction 1 9 Disconnecting the power 1 9 Disconnecting the power The power to the AxiChrom column AxiChrom Master and or the system can also be shut off in an emergency by disconnecting the power supply using the system power switch Note that disconnecting the power this way from an AKTAprocess system will cause valves to close immediately and not in a proper sequence Ensure that the system and column is ful
36. the slurry hose is clean stop J the pump Close the slurry tank valve and press CONFIRM Pressure 0 05 bar Adapter stopped Media valve closed CONFIRM 1 Keep the valve on the slurry tank open N Start the pump at the stated flow through the rinse tube and back into the slurry tank 3 When the slurry tube has been rinsed free from slurry e Stop the pump e Close the valve on the slurry tank 4 When all above conditions are fulfilled and their boxes have been checked press CONFIRM to proceed to the next step Step 11 Open the bottom mobile phase valve Packing progress Open the bottom mobile phase 11 15 ea Capto Q Adapter position 117 cm opanthe mobie Top mobile phase should be kept closed Press CONFIRM to start the packing Pressure 0 05 bar Adapter stopped Media valve closed HELP ABO CONFIRM 128 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 4 Packing using the Master Wizard 1 Ensure that the Top mobile phase valve is kept closed 2 Open the Bottom mobile phase valve and check the box in the dialog 3 Press CONFIRM to proceed with packing Step 12 Consolidation phase Packing progress 12 15 6 05 romaning Dialek Adapter position 15 1 cm Adapter speed GO cmh Time to calculated bed detection 4 05 Pressure 0 14 bar Adapter moving Media valve closed The consolidation starts An approximate total time to the low bed height l
37. waeeccessessssssssssssssssssssccsssssssssssssssssssesssssssssssssssssssessesssssssssssessssssseesessssssees 138 Evaluating column performance s ss seessssssesssssssessssssessssessessssesesessessesessessesesess 141 6 1 Choice of test samples for COLUMNS waacecccssssssssssssssssesseseccssssssssusssssssssesssssesssssssessessecsessssssssseesees 141 6 2 HETP and asymmetry calculation oe eescesssssscsssssssseesssseseccssssssssusssessessecssssesssussseessssessesesssassneesees 142 After the Packing aise eee aha icici cies estab esis Lien kadina daa kaikien riaan ha 144 EL Clegning in plac CIP sissioni aaan iin 144 Le Sanitization Nn pli snae AAAA RNR 144 T3 Unpacking the coluisse na ann A NRN 145 7 3 1 Short description of the unpacking procedure accceesesssssssssssssssssssssssssssssssssssssssseesesssssssssssesess 146 7 3 2 Creating a UNICORN method esssssesessssssscssssssssssssssssssssesssssssssssssssssssseccesssssssssssssssssesesssssssssssssseeess 146 7 3 3 Unpacking acrylic column tubes 150 7 3 4 Unpacking stainless steel column tubes 162 LA System shutdown proc dure suinosinianianaianinnuia anunn aia 163 Parts list and didor amS scsi aan steer eee eee 167 A 1 Part numbers and NAMES oo ssssssssessssssessssssssssssnsssssnssssssesssssnssssssssssestssseeseseeceneeeeecseceeceeneeccecceneeeeeeeee 167 A 2 Columns with any bed support Mate rich o eessssessseesssssesssssnssssssnsssseessssssseceseseceeeeeceeeeeneceeee 172 A 3 Column
38. will have to be recreated according to the instructions in Section 3 7 1 Start Master and cali brate column on page 75 3 Disconnect the Master power cable 4 Disconnect the motor power cable 5 Disconnect the motor feedback cable 6 Disconnect the pneumatic air supply 7 Optionally dismount the Master pressure transmitter from the flow path 166 AxiChrom 300 1000 columns User Manual 28 9562 89 AD Appendix A Parts list and diagrams Note A Parts list and diagrams A 1 Part numbers and names Diagrams showing AxiChrom 600 main column components The appearance may differ between different column sizes The table below lists the main parts of the column See product documentation for full listing 104 Tube holder 400 to 1000 105 Tube holder bracket 106 Holder 109 Locking part 111 Fastener adaptor 115 Fastener Tube holder 300 to 600 116 Flushing connection 117 Flushing connection nut 126 Plate 127 Shock absorber lower 128 Top plate 129 Level display 132 Tube Roller asm 133 Flexible tube 3 4in TC25 134 Lid support complete 136 Locking pin asm 137 Flushing Inlet Tubing L1000 143 Hex Head Screw ISO 4017 M6x25 A4 70 144 Screw M6S H M5X14 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 167 A Parts list and diagrams A 1 Part numbers and names
39. with priming solution and the adapter moves to the priming position If the adapter does not move it is already in the correct position Note With stainless steel column tubes it is not possible to see when liquid has passed the priming groove Verify instead that the adapter is in the top position and that buffer flows out of the top mobile phase outlet before clicking Continue When prompted to do so increase the pressure in the column by tightening the membrane valve to approximately 0 3 bar open the Seal flush valves and then click Continue This will happen when the adapter is in priming position above the priming groove the column has been filled and priming solution comes out in waste through the system and through the Seal flush valves and the flow is lowered Result The adapter then moves down at 40 cm h to a position under the priming groove expelling the air below the scraper seals as the adapter passes the groove During the priming direct the Seal flush valve to waste or to a vessel AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 4 Priming the column 4 4 4 Using UNICORN method 5 The adapter stops below the priming groove When prompted to do so open the membrane valve slowly to release pressure in the column close the Seal flush valves and click Continue
40. 0 Note Default values for variables in the Method Wizard may be changed to optimize an Intelligent Packing method Changes using the variables page in the Method Editor module should only be made by advanced users 1 Start the Method Wizard in the Method Editor window and choose a system in the pop up window AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 2 Creating a UNICORN method 2 Select whether to create a method for Column Packing Packing Test or both combined Saiid Wieus fur syo fin AAT iosusees Atlus souio Innige Packing Coke Packing w h Packing Test AxiChrom lt Setting Description Column Select AxiChrom lt xxx gt lt yyy gt xxx column diameter in millimeter yyy max bed height for short and long column tube 300 or 500 mm respectively The column size can be found on a label located on the lower front part of the frame of the column Column material Select the material of the column tube Show medium type Select the appropriate medium type Medium Select the appropriate medium For custom medium see 5 2 1 Creating a packing method based on custom medium Set Default AxiChrom 300 1000 columns User Manual 28 9562 89 AD Sets the default values used in the Method Wizard 111 5 Packing the column 5 2 Creating a UNICORN method 112 Click Next to display the Packing Parameters 1 Method Wizard page Select
41. 0 or 20 um 401 Distributor bottom 600 402 Distributor adaptor 600 403 Bed support screw bottom 300 600 404 Bed support screw adaptor 300 600 405 Distributor adaptor ring 600 406 Snap ring 600 409 Fastener bottom 411 T piece 3TC ASME BPE DT 18 1 2in SF5 412 Pressure Transmitter 413 O ring 26 7x1 78 414 O ring 600x10 415 O ring 556 86x6 985 416 O ring 506 86x6 985 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 169 A Parts list and diagrams A 1 Part numbers and names 417 O ring 30x3 5 421 Hex Socket Head Cap Screw ISO 4762 M8x25 A4 70 422 Washer ISO 7089 8 4x16x1 6 A4 423 Parallel Pin ISO 2338 6m6x16 A2 431 Hex Nut ISO 4032 M10 A4 70 432 Washer ISO 7089 10 5x20x2 A4 501 Tube Mobile Phase bottom 502 Tube Slurry inlet outlet bottom 503 Tube Rinse bottom 504 Connector top inlet 505 End cap 506 TC clamp for TC 25 507 TC gasket TC25 ID15 EPDM 508 Inner Connector Rinse only used with PP tubes on 300 600 columns 509 O ring only used with PP tubes on 300 600 columns 511 Top Inlet outer body 512 O ring 16x3 EPDM70 513 Parallel Pin ISO 2338 4M6x16 A2 516 Valve body outer 517 Valve body inner 518 O ring holder 519 Piston 521 Pneumatic cylinder 537 Washer 541 O ring 125x5 542 Top inlet 543 Valve 170 AxiChrom 300 1000 columns User Manual 28 9562 89 AD A Parts list and diagrams A 1
42. 00 2230 2340 2630 2650 Long 2720 2750 2860 3150 3170 Height D when adapter is at max bed Short 1740 1760 1870 2160 2170 aoe Long 2200 2220 2330 2620 2630 Min height delivery height Short 1450 1460 1480 1590 1880 1890 Long 1710 1720 1740 1850 2140 2150 Max slurry filling height Short 2040 2060 2185 2335 2490 Long 2555 2580 2700 2850 3005 Max operating height D during priming Short 2060 2080 2190 2480 2490 Long 2580 2600 2710 3000 3010 Max adapter stroke height for filling Short 570 578 Long 830 838 1 Refers to the height height of casters excluded of the column when the adapter is in its highest position for example for working and service 2 Bed height 100 to 300 mm 3 Bed height 100 to 500 mm 2 3 3 Operating conditions Parameter Value Maximum operating pressure 1 4bar Operating temperature 2 2 C to 30 C Operating pH 3 1to 14 1 Refers to the maximum allowed pressure over the bed during normal chromatographic operation 2 See Section 3 9 Ambient temperature changes on page 86 3 Due to corrosion low pH values are unsuitable for solutions containing chlorides See Section 2 5 Chemical resistance on page 36 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 33 2 Specifications and characteristics 2 3 Primary specifications 2 3 4 Pressure and time out limits 2 3 4 Pressure and time out limits Descripti
43. 2 15 Prime the bottom mobile phase Close the top mobile phase J Open the bottom mobile phase and i Girect it towards waste Adapter position 30 0 cm The adapter will start at a high speed to prime the bottom screen expelling Rquid through the bottom mobile phase Pressure 0 03 bar Press CONFIRM to continue Adapter stopped Media valve closed 1 Close the Top mobile phase valve 2 Open the Bottom mobile phase and direct its tubing to waste 3 Press CONFIRM when all the conditions have been fulfilled and all the boxes are checked The adapter will then start moving at a high speed to prime the bottom bed support expelling liquid through the Bottom mobile phase 106 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 4 Priming the column 4 4 5 Using Master wizard Step 14 wait for the adapter to move down Priming progress Moving the adapter down 13 15 aa 4 42 roaming Adapter commanded postion 1 0 cm LLL Adapter position 24 5 cm ERE NE SON Moving the adapter down Pressure 1 23 bar Adapter moving Media valve closed The adapter now moves down to bottom position The remaining time is displayed in a time bar Press PAUSE to stop the movement temporarily or to terminate the priming wizard Step 15 close the bottom mobile phase valve Priming progress lose the bottom mobile phase 14 15 Close the bottom mobile phase Close the bottom mobile phase flow pa
44. 300 1110 x 620 1470 2330 699 710 450 100 to 500 1110 x 620 1730 2750 749 760 600 100 to 300 1180 x 780 1570 2340 818 835 600 100 to 500 1180 x 780 1830 2860 883 900 800 100 to 300 1470 x 1080 1720 2630 2122 2150 800 100 to 500 1470 x 1080 1980 3150 2212 2240 1000 100 to 300 1720 x 1300 1875 2650 2517 2560 1000 100 to 500 1720 x 1300 2135 3170 2637 2680 1 Weights are given for columns with plastic bed support stainless steel bed support AxiChrom 300 1000 columns User Manual 28 9562 89 AD 193 E Column and AxiChrom Master weights and dimensions Column weights and dimensions stainless steel column tube The table below lists column tube weights and the weights of empty columns and crate column for columns with stainless steel tubes All weights are given in kg Column Column tube Empty column Crate column 300 short tube 73 431 437 591 597 300 long tube 89 454 460 619 625 400 short tube 92 481 490 641 650 400 long tube 113 508 518 674 683 450 shorttube 106 730 741 910 921 450 long tube 130 786 797 971 982 600 short tube 178 864 881 1059 1076 600 long tube 209 930 947 1135 1152 800 short tube 351 2231 2259 2581 2609 800 long tube 394 2318 2346 2678 2706 1000 short tube 584 2738 2781 3178 3221 1000 long tube 637 2841 2884 3291 3334 1 Weights are for columns with plastic bed support stainless steel bed support 2 Weights are for columns with plastic bed support
45. 890 450 100 to 500 2180 1090 1480 185 934 945 600 100 to 300 1875 1100 1395 195 1013 1030 600 100 to 500 2135 1100 1395 205 1088 1105 800 100 to 300 2190 1440 1800 350 2472 2500 800 100 to 500 2450 1440 1800 360 2572 2600 1000 100 to 300 2240 1650 2040 440 2957 3000 1000 100 to 500 2500 1650 2040 450 3087 3130 Master 1300 770 700 65 140 1 Weights are given for columns with plastic bed support stainless steel bed support Note Some crate weights are estimates 192 AxiChrom 300 1000 columns User Manual 28 9562 89 AD E Column and AxiChrom Master weights and dimensions Appendix E Column and AxiChrom Master weights and dimensions The actual weight and dimensions for the individual column is given in the documentation Typical weights and dimensions for the different column sizes are listed in the table below The min and max heights refer to the height of the column with the adapter in its lowest position for transportation and with the adapter in its highest position for example for maintenance work Column weights and dimensions acrylic column tube Column Column bed Footprint Min Max Weight of empty diameter heights length x width height height column mm mm mm mm mm kg 300 100 to 300 1110 x 520 1450 2220 414 420 300 100 to 500 1110 x 520 1710 2720 434 440 400 100 to 300 1110 x 600 1425 2220 451 460 400 100 to 500 1110 x 600 1685 2720 471 480 450 100 to
46. Always use protective glasses and other personal protective equipment appropriate to the current application to ensure per sonal safety during operation WARNING Disconnection of pressurized air supply tubes can cause loud noises Ear protection is recommended Note No ear protection is required during normal operation The noise from AxiChrom is low below 55 dB Installing and moving the columns and Master WARNING Move transport crates Ensure that the forklift has capacity to safely lift the crate weight Ensure that the crate is properly balanced so that it will not accidentally tip when moved WARNING Move column Ensure that the column s center of gravity is well balanced over the forklift s lifting arms otherwise the column may tip off the forklift WARNING All electrical installations must be performed by authorized personnel only AxiChrom 300 1000 columns User Manual 28 9562 89 AD 15 1 Introduction 1 6 Safety precautions CAUTION Ensure that all tubing hoses and cables are placed so that the risk for tripping accidents is minimized NOTICE Disconnect all tubing hoses and cables before moving the column O gt System operation WARNING The working pressure of the column should never exceed 4 bar otherwise there is a risk of personal injury and damage to the col umn Always use appropriate pressure alarms pressure vents or ru
47. C Machinery Directive MD 2006 95 EC Low Voltage Directive LVD 2004 108 EC ElectroMagnetic Compatibility EMC Directive 97 23 EC Pressure Equipment Directive PED 10 AxiChrom 300 1000 columns User Manual 28 9562 89 AD International standards 1 Introduction 1 4 Regulatory information This product is designed in accordance with the requirements of the following standards and regulations Standard EN 61010 1 IEC 61010 1 UL 61010 1 CAN CSA C22 2 No 61010 1 Description Safety requirements for electrical equip ment for measurement control and lab oratory use Notes EN standard is harmonized with EU directive 2006 95 EC EN 61326 1 Emis sion according to CISPR 11 Group 1 class A EMC emissions and immunity require ments for electrical equipment for mea surement control and laboratory use EN standard is harmonized with EU directive 2004 108 EC EN ISO 12100 Safety of machinery General principles EN ISO standard is for desgn Risk assessment and risk reduc harmonized with tion EU directive 2006 42 EC ASME BPVC Boiler and Pressure Vessel Code BPVC Applies only to VIII div 1 stainless steel columns ASME BPE Bioprocessing equipment UL 508a UL standard for Safety for Industrial Control Panels EMEA 410 01 All wetted polymer and elastomeric parts CPMP Note are animal origin free or comply with the conditions in the standard and are als
48. Chrom Master is equipped with an EMERGENCY STOP button which can be used to stop the movement of the adapter in case of an emergency during operation of the column When the EMERGENCY STOP button is pressed the adapter movement stops The process that is in progress in AxiChrom Master for example an Intelligent Packing will be aborted and the interface will display the MAIN MENU The EMERGENCY STOP button locks in place until it is released manually The EMERGENCY STOP button is located on the AxiChrom Master The EMERGENCY STOP label is yellow with black text and the button is red ae i WARNING Ensure that the EMERGENCY STOP button can always be reached while working with the column 20 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1 Introduction 1 7 Emergency stop procedures WARNING The EMERGENCY STOP button will not shut off the electrical power to the motor of column Use the power switch on the connector panel of AxiChrom Master to shut off the electrical power EMERGENCY STOP buttons on AKTAprocess systems The EMERGENCY STOP buttons on the system must be used in case of an emergency when the pumps have to be stopped There are two EMERGENCY STOP buttons on an AKTAprocess system one on each side and only one needs to be pressed in order for the pumps to stop Refer to the Operating instructions for more information about the emergency procedures for AKTAprocess 1 8 Resetting the
49. D 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes 1 Start the pump upflow from the Bottom mobile phase using the flow stated in the dialog 2 When the column is empty and the pump has been stopped press CONFIRM to continue to the next step Step 15 Rinse the media valve and slurry tubing npacking progress Rinse 15 16 awam Capto Q Rinse Adapter position 1 0 cm Keep the slurry tank valve open Start the pump at 503 Uh through the J Rinse hose and further into the slurry tank When the sluny hose is clean stop the pump Close the slurry tank valve and press CONFIRM Pressure 34 mbar Adapter stopped Media valve closed CONFIRM Rinsing the Media valve will clean the valve and slurry tube from media after the unpack ing 1 Ensure that the valve on the slurry tank is open 2 Start the pump at the flow stated in the dialog through the rinse tube and back into the slurry tank 3 When the slurry tank tube is emptied of gel stop the pump 4 Close the valve on the slurry tank 5 Check the boxes and press CONFIRM AxiChrom 300 1000 columns User Manual 28 9562 89 AD 161 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes Step 16 Exit the wizard amwa Capto Q The unpacking has been completed Please measure the slurry concentration before repacking the column The unpacking is now completed Measure the slur
50. GE Healthcare Life Sciences AxiChrom 300 1000 columns User Manual Table of Contents Table of Contents L sos 0 nero eee nee ecg nee i er ee RU ere err erent arr tt neler a reer Nes cree 5 1 1 General introduction of 5 1 2 Scope of this User Manual 6 1 3 Important user information is 8 14 REGION IMPORT ON esiisa n ai RAR 10 1 5 intended use of the COLUMNS oeceeeccccccessssssssssssssssssssseeeesseeeesseeeesssesessseesessesessssesssssssessssesssssessssnsessnats 12 1 6 Safety precautions u 13 1 7 Emergency stop procedures 20 1 8 Resetting the system after an emergency stop ai 21 1 9 DISCONNECTING th POWT vecseecsccsssssssssssssssssusssscssccsssssssssusssseseccesssssssussssssseseesesssssssssssesessessesenssuassees 22 MMO SSCS AET EE EA OEE O EEE EETA E ES 22 2 Specifications and characteristics cccscesesssscssecessesssesscssseesseessssseseseeseeees 26 21 Principles Of column FUNCTION oiaseesnanirsessnmnannenasssunnnanhissaninnihs 26 2 2 Overview of the column and parts csssssssessssssccssssssssssssessesssesscsssssssssssecsessessssssssssussssssscessssenssees 28 2 3 Primary Specificato NS sszsssiasssvssssvusdeccessseicovtelastssoacSeisecdenvatisaas oonssnscoovaetaveieiseebdbsnobaevbansshsabeidbvnealenaoen 32 2 3 1 Weights volumes and related dimensions 32 2 3 2 Heights in different operating states 33 2 3 3 Operating conditions cssssssse
51. MMC Any salt solu 1 16 60 tion Capto S Water 1 10 60 0 20 EtOH Capto S Any salt solu 1 20 60 tion CM Sepharose Fast Flow Water 1 15 60 0 20 EtOH SP Sepharose Big Beads Water 1 18 60 0 20 EtOH SP Sepharose Big Beads Any salt solu 1 20 60 tion SP Sepharose Fast Flow Water 1 15 60 0 20 EtOH SP Sepharose Fast Flow Any salt solu 1 15 60 tion SP Sepharose High performance Water 1 15 30 0 20 EtOH SP Sepharose High performance Any salt solu 1 17 30 tion SP Sepharose XL Water 1 15 60 SP Sepharose XL Any salt solu 1 15 60 tion Hydrophobic Butyl S Sepharose 6 Fast Flow 10 20 EtOH 1 15 60 interaction Butyl Sepharose High Performance 10 20 EtOH 1 15 30 Phenyl Sepharose High Performance 10 20 EtOH 1 15 30 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 185 B Packing buffer information Packing buffer Packing factor Packing velocity cm h Metal Chelating Sepharose Fast Flow Water 1 15 60 affinity 0 20 EtOH IMAC Sepharose 6 Fast Flow Water 1 15 60 0 20 EtOH Ni Sepharose 6 Fast flow Water 1 15 60 0 20 EtOH Size Superdex Water 1 15 30 exclusion 0 20 EtOH Custom Custom default settings Custom 1 0 30 Unpacking values All values are in cm h Media Expand flow Unpacking Unpacking Adapter unpacking expand speed flow speed Capto MabSelect Fast Flow 75 60 30 60 XL Big Beads High Performance Superdex 40 cycle 1 30 30 60
52. North America Frequency Supply voltage frequency Max Power Consumption Maximum power consumption Enclosure Enclosure rating according to EN 61326 1 Largest Motor Current FLA Highest current used by the motor Full Load Ampere Max Current Highest current used by the AxiChrom Master Pneumatic Supply Required pressure for pressurized air supply Short Circuit Rating Maximum tolerated short circuit current Protection Class Protection class rating according to EN60529 24 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1 Introduction 1 10 Labels Symbols used in safety labels Label Meaning Warning Read the user documentation before using the system Do not AN open any covers or replace parts unless specifically stated in the user documentation The system complies with the requirements for electromagnetic compliance EMC in Australia and New Zealand C The system complies with applicable European directives Labels concerning hazardous substances Label Meaning This symbol indicates that waste electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected sep arately Please contact an authorized representative of the manufacturer for information concerning decommissioning of equipment Be This symbol indicates that the product contains hazardous materials in Lay excess of the limits established by the Chinese stan
53. ORN 6 Choose an appropriate result file folder result name and save the method 44 4 Using UNICORN method The Media valve column and system are primed using UNICORN and KTAprocess The priming sequence using a UNICORN method is the same as when priming using the Master wizard from Step 5 The difference is that during UNICORN control manual inter actions are minimized to control the Seal flush valves and to perform some confirmations The KTAprocess system controls other valves flows and rinsing procedures Preparations before running the method Connect the column using the automatic configuration for AktaProcess as described in Section 3 6 Column and system setup on page 68 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 97 4 Preparations before packing the column 4 4 Priming the co 4 4 4 Using UNICO 98 umn RN method WARNING Ensure that nobody is present within the safety zone when the adapter is moving The operator in charge is responsible for keeping the safety zone free from unauthorized personnel and unnecessary equipment Running the method 1 4 Start the priming method Result The system performs a wash procedure The Media valve and the slurry tubing tank are then primed if selected in the Method Wizard dialog Click Continue when prompted to do so when the stated conditions have been fulfilled Result When the Media valve has been primed the column is filled
54. Packing velocity Packing buffer factor cm h Affinity Blue Sepharose Fast Flow Water 1 15 60 0 20 EtOH Heparin Sepharose 6 Fast Flow Water 1 15 60 0 20 EtOH MabSelect Water 1 15 60 0 20 EtOH MabSelect SuRe Water 1 151 60 0 20 EtOH MabSelect SuRe LX Water 1 151 60 0 20 EtOH MabSelect Xtra Water 1 15 60 0 20 EtOH 1 When calculating the amount of media to be packed use the Compression Factor 1 1 instead of the Packing Factor above which is used during packing AxiChrom 300 1000 columns User Manual 28 9562 89 AD 183 B Packing buffer information Packing Packing velocity Packing buffer factor cm h Anion Capto adhere Water 1 10 60 exchange 0 20 EtOH Capto DEAE Water 1 10 60 0 20 EtOH Capto DEAE Any salt solu 1 15 60 tion Capto Q Water 1 10 60 0 20 EtOH Capto Q Any salt solu 1 15 60 tion DEAE Sepharose Fast Flow Water 1 15 60 0 20 EtOH Q Sepharose Big Beads Water 1 18 60 0 20 EtOH Q Sepharose Fast Flow Water 1 15 60 0 20 EtOH Q Sepharose High Performance Water 1 15 60 0 20 EtOH Q Sepharose XL Water 1 15 60 0 20 EtOH 184 AxiChrom 300 1000 columns User Manual 28 9562 89 AD B Packing buffer information Packing Packing velocity Packing buffer factor cm h Cation Capto MMC Water 1 13 60 exchange 0 20 EtOH Capto
55. Part numbers and names 601 O ring 14x3 602 O ring 48x3 5 603 O ring 21 2x3 604 O ring 27x4 607 O ring 48x3 5 611 O ring 48x3 5 612 O ring 27x4 614 O ring 48x3 5 616 O ring 14x1 78 617 O ring 14x1 78 618 O ring 8x3 624 O ring 14x3 628 O ring 565x11 641 Scraper 710 Bellows 711 Screw and gear 712 Adapter rod with level scale 713 Adapter rod 801 Hex Head Bolt M20x110 802 Hex Head Bolt M24x300 803 Hex Nut M24 811 Bottom backing plate 812 Adapter backing plate 813 Lid 814 Adapter stop 904 Level display label AxiChrom 300 1000 columns User Manual 28 9562 89 AD 171 A Parts list and diagrams A 2 Columns with any bed support material A 2 Note 172 Columns with any bed support material e The numbers correspond to position numbers in Material Conformity and Spare Parts Grey circles indicate parts which retain pressure or are in contact with process fluids aaa teeny l a te t LS N 8 7 132 Fae D An t A a A Oe A 133 7 oD Saz Parts 411 and 412 are mounted on the system Part 133 Storage solution fill hose is dismounted after commissioning AxiChrom 300 1000 columns User Manual 28 9562 89 AD A Parts list and diagrams A 2 Columns with any bed support m
56. TC50 ID 22 1 TC50 ID 34 8 28 9381 76 TC gaskets Article name Code no TC Gasket 25 ID 10 5 For tube ID 7 75 28 9381 86 TC Gasket 25 D 12 For tube ID 9 4 28 9381 87 TC Gasket 25 D 15 8 For tube ID 15 75 28 9381 88 TC Gasket 50 22 1 For tube ID 22 1 28 9668 86 TC Gasket 50 34 9 For tube ID 34 8 28 9381 89 Clamps Article name Code no Clamp TC25 28 9381 90 Clamp TC50 28 9381 91 188 AxiChrom 300 1000 columns User Manual 28 9562 89 AD C Accessories and additional literature Wheel kits AxiChrom 300 wheel kit 28 9381 92 AxiChrom 400 wheel kit 28 9381 93 AxiChrom 450 wheel kit 28 9554 29 AxiChrom 600 wheel kit 28 9381 94 AxiChrom 800 wheel kit 28 9381 96 AxiChrom 1000 wheel kit 28 9381 97 Grounding kit Article name Code no Grounding kit 18 1157 87 Toolkit Article name Code no Tool kit Axichrom 300 1000 28 9382 03 Plastic mallet 28 9820 32 Removal tool 28 9820 31 Safety valves Article name Code no Safety Valve 4bar ID 9 4 28 9382 00 Safety Valve 4bar ID 15 75 28 9382 01 Safety Valve 4bar ID 22 1 28 9382 02 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 189 C Accessories and additional literature Rupture disc and holder Article name Code no Rupture disc kit 4 2 bar for AxiChrom 300 600 29 0139 97 contains one special T junction and one rupture disc Rupture disc 4 2 bar TC50 29 0139 98
57. Use the arrow buttons to reduce the adapter speed if the adapter comes too close to the bed Make sure the distance between the bed and the adapter is at least 2 cm The adapter speed range depends on the media that is unpacked The largest range is from 30 cm h to 240 cm h e Ifthe adapter movement stops due to the electric current in the motor rising too high the method is aborted and an alarm is raised The unpacking must then be restarted from the beginning or be continued manually e Ifthe pressure exceeds the maximum allowed the adapter movement will temporar ily stop until the pressure has dropped When unpacking bead size d gt 50 um e g Capto Fast Flow or MabSelect media the adapter will stop if the pressure reaches 0 5 bar and start to move again when pressure falls below 0 2 bar When unpacking bead size d lt 50 um e g High Performance media the adapter will stop if the pressure reaches 1 0 bar and start to move again when pressure falls below 0 5 bar The pressure will decrease automatically when the adapter movement has stopped If the pressure does not drop automatically ensure that The valve on the slurry tank is open The slurry tube is not clogged The flow is not too high AxiChrom 300 1000 columns User Manual 28 9562 89 AD 159 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes 160 e Ifthe adapter has stopped due to high pressure it is not possible to chan
58. acrylic column tubes Not for use with columns contaning wetted components of stainless steel Abstract Services CAS American pH in these solutions depends on the pH of the buffer which can vary between 3 and 13 On stainless steel column parts only Exposure of other column parts to ethanol at higher For columns containing wetted stainless steel components max 1 0 M NaCl is recommended For additional important information see Section 2 5 1 Resistance to sodium chloride on page 39 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 2 Specifications and characteristics 2 5 Chemical resistance 2 5 1 Resistance to sodium chloride Avoid using the following chemicals in AxiChrom columns e Extreme oxidizers e Fluorine and halogenated compounds e Chlorinated solvents such as methylene chloride e Esters e Aromatic hydrocarbons such as toluene e Alcohols at concentrations higher than those specified in the table above e Salt in combination with pH below 4 Always wash the column with at least five col umn volumes of pH neutral solution water between and after use of salt and low pH buffers 2 5 1 Resistance to sodium chloride Depending on configuration Axichrom columns can be equipped with wetted compo nents of stainless steel and must therefore be appropriately maintained when exposed to sodium chloride NaCl during chromatographic processes Since the bed supports are made of stainless steel and have a
59. al use In a domestic environment it may cause radio interference in which case the user might be required to take appropriate measures Note This equipment complies with FCC part 15 2004 Radio frequency device subpart B Unintentional radiators Class A Operation is subject to the following two conditions 1 This equipment may not cause harmful interference 2 This equipment must accept any interference received including interfer ence that may cause undesired operation Grounding the column The column should always be connected to ground using a ground cable kit Use the connection point on the inside of the column stand to ground the column to a suitable grounding point The illustration below shows the connection point for protective ground cable on the column frame Figure 3 1 Connection point on the column frame for protective ground cable AxiChrom 300 1000 columns User Manual 28 9562 89 AD 65 3 Getting started 3 4 Connections Compressed air requirements It is important for personal safety and safe operation to use the correct pressure and quality of compressed air for the pneumatic valve control The basic requirements are e Free of oil and particles e 30 C dew poin
60. ance read from the level scale on the adapter rod Observe that the scale has increasing figures running downwards x s 8 Note The level scale indicator differs between columns with different bed support material plastic or stainless steel Make sure that the correct level scale indicator is fitted 2 Press Set to change the dialog value Last known adapter position if it differs from the actual adapter position read from the level scale Set the value to a tolerance of one decimal in centimeters 3 Press CONTINUE to enter the MAIN MENU AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 7 Using the Master 3 7 3 Column and Master setup Verify the factory calibration value for the level display 1 Connect the column and the Master 2 Start the Master 3 Verify that the adaptor value in the master correspond to the value on the Level display label 4 Check the factory Master calibration value cm in the functional test records in the column product documentation 5 Move the adaptor upwards using a low speed 30 cm h to the end of the stroke Use the current limitation to stop the adaptor 6 Verify that the adaptor height on the level display scale corresponds to the test record Height difference lt 0 2cm Master configuration Description To access the configuration wizards press the Configuration button on the Select column page lt BACK Use the configur
61. answer the questions and enter a result name Ensure that the Master is in MAIN MENU before continuing Click Start to start the method When prompted to open the slurry tank and then click Continue Keep the media homogeneously suspended throughout the entire fill step Result The column is filled automatically with slurry from the slurry tank by raising of the adapter The filling stops automatically and the Media valve closes when correct amount of slurry is filled into the column The maximum filling speed is 300 cm h After the filling the system rinses the Media valve from slurry back into the slurry tank When the slurry tube is free from slurry click Continue Close the slurry tank valve and click Continue Result The bed consolidates as the adapter moves down in the column A warning dialog is displayed three or six minutes before the calculated consoli dated bed is reached depending on media The warning dialog should be closed before detection of the bed occurs AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 3 Packing using a UNICORN method 7 Pay attention as the adapter approaches the consolidated bed surface When the bed is detected a confirmation dialog appears Note When the adapter comes into contact with the consolidated bed there will be a lag default 2 seconds to sense if the detection has occurred or not When the detection has occurred the operator confirms that the detec
62. any substances used in the system are hazardous or toxic these must be highlighted under 6 3 1 hereby confirm that the equipment has been thoroughly cleaned whether or not the equipment has been in contact with toxic and hazardous substances and that the equipment has been filled with a suitable solution have also at tached a description of the cleaning method Storage solution in equipment Signed Position Document number 04 0051 11 Edition AA WE imagination at work AxiChrom 300 1000 columns User Manual 28 9562 89 AD 195 For local office contact information visit www gelifesciences com contact GE Healthcare Bio Sciences AB Bj rkgatan 30 751 84 Uppsala Sweden www gelifesciences com axichrom imagination at work GE imagination at work and GE monogram are trademarks of General E Company lectric Akta AKTAprocess AxiChrom Capto DropDesign HiScale MabSelect MabSelect SuRe MabSelect Xtra Sepharose Superdex and UNICORN are trademar GE Healthcare companies 2009 2012 General Electric Company All rights reserved First published Sept 2009 All goods and services are sold subject to the terms and conditions of sa the company within GE Healthcare which supplies them A copy of these and conditions is available on request Contact your local GE Healthcare sentative for the most current information s of e of terms repre UNICORN Any use of this software is su
63. aterial Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids CN a on W on i d CaA S ey Part 536 is not included in newer inlet outlets AxiChrom 300 1000 columns User Manual 28 9562 89 AD 173 A Parts list and diagrams A 2 Columns with any bed support material Columns with acrylic tubes Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids eH 109 174 AxiChrom 300 1000 columns User Manual 28 9562 89 AD A Parts list and diagrams A 2 Columns with any bed support material Columns with stainless steel tubes Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids Mounted on system Vow he Storage solution fill hose Dismounted after commissioning Mounted on system Parts 411 and 412 are mounted on the system Part 133 Storage solution fill hose is dismounted after commissioning AxiChrom 300 1000 columns User Manual 28 9562 89 AD 175 A Parts list and diagrams A 2 Columns w
64. ation wizards to e run startup tests and access the Panel view only accessible by a GE Healthcare service operator e view system settings only accessible by a GE Healthcare service operator e view system information and specifications AxiChrom 300 1000 columns User Manual 28 9562 89 AD 83 3 Getting started 3 7 Using the Master 3 7 3 Column and Master setup System Info hrom master system int Revisions and serial numbers DriveLogix 730 PowerFlex 700S 2 400V PLC Firmware rev 16 20 VFD Firmware rev 4 1 Serial no 20174946 Application rev 2 10 1788 Module Profibus slave Operator Panel Application rev 2 10 CONTINUE gt This dialog displays serial numbers and firmware revisions Press CONTINUE to proceed Note The above image is an example Other firmware and application revisions may apply Necessary adjustments If Then The column is new e Adjust the tightening torque of the bolts on the tie rods see Tightening torques on page 87 The column has been Adjust the tightening torque of the bolts on the tie rods moved from a room see Tightening torques on page 87 with a different temper ature The room temperature has changed significantly The column tube or Contact GE Healthcare to have a service technician per other major parts have form an adapter position calibration been replaced 84 AxiChrom 300 1000 columns User Manual 28 9562 89 AD Leakage test 3
65. ay than described in the user documentation If you do you may be exposed to hazards that can lead to personal injury and you may cause damage to the equipment 8 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1 Introduction 1 3 Important user information Intended use The AxiChrom family of process columns has been designed for low pressure chromato graphic separation of biomolecules such as proteins peptides and oligonucleotides in GMP regulated environments AxiChrom columns are intended for production use only and should not be used for diagnostic purposes in any clinical or in vitro procedures The columns are not suitable for operation in a potentially explosive atmosphere or for handling flammable liquids If the columns are used for purposes other than those specified in the user documentation safe operation and the protection provided by the system may be impaired Safety notices This user documentation contains WARNINGS CAUTIONS and NOTICES concerning the safe use of the product See definitions below Warnings WARNING WARNING indicates a hazardous situation which if not avoided could result in death or serious injury It is important not to proceed until all stated conditions are met and clearly understood Cautions CAUTION CAUTION indicates a hazardous situation which if not avoided could result in minor or moderate injury It is important not to pro ceed until all stated condition
66. bject to GE Healthcare Standard So End User License Agreement for Life Sciences Software Products A copy Standard Software End User License Agreement is available on request GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare UK Limited Amersham Place Little Chalfont Buckinghamshire HP7 9NA UK GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan 28 9562 89 AD 06 2012 ftware of this
67. centration 91 4 4 Priming the column s sssssesssssssssssesssrorrossssssrssessrrosessee m 93 4 4 1 Short description of the priming procedure ssssssssssssssssssssssssssssssssssssssssnansenesneneennneeeeeeeee 93 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Table of Contents o a o m AAP Wetting plastic bed SUPDOMES soiiinniiroriiisiiiuenan ia 94 4 4 3 Creating a UNICORN Method uncceccssssssssssssssssssssscsssssssssssssssssssessssssssssssusssssssseesssssssssuussssessseesesssssnees 94 4 4 4 Using UNICORN method 97 445 Using Master WOA ccccsisssscessasecsesscscscssoistessstbasesavovssccistetnsaatesssvavsussoessescessusssshceusievestececespecsstsstdtans 99 Packing the colui sorsi e E eens 108 51 Intelligent Packissa ANN ANNANN EE 108 52 Creatingia UNICORN method srsssssssissscccsescosssssiissseassscactexveusaisussbennsvevsisetordensistalide ries subionteobessttGioee 110 5 3 Packing using a UNICORN method o ccccceccssssssssssssssssssesssssessssssessssssessnnssssnnnsssiinsssnnaissannnnness 117 5 4 Packing using the Master Wizard ssssssssssessccsccsssssssssussessesccsssssssssussssssssscssssssssssssuessssssessssesssess 120 5 5 Creating custom packing methods in the Master ccsssssssssssesssssessssssessssssssssssesssssesssanseses 135 5 5 1 Packing stainless steel COLUMN tubes wececcccsssssssssssssssssesssssssccssssssssssssssssssecsssssssssssssesesseeesessssssees 137 5 5 2 POSt packing modifications
68. continue the intelligent packing using the predefined conditions i Keep the media homogenously suspended Adapter speed 2 _4 Pressure 0 25 bar adjustment tenia aladspeed 200 PAUSE e The adapter moves upwards e Keep the media homogeneously suspended e The adapter stops and the Media valve closes automatically when the filling is finished e Ifthe pressure falls below 0 30 bar g the filling will stop until the pressure has risen above 0 25 bar again e The adapter speed can be controlled using the arrow buttons If the adapter stops repeatedly the slurry concentration is too high and the adapter filling speed needs to be lowered The speed range limit is 100 to 300 cm h e Ifthe filling for some reason needs to be ended earlier press the END FILLING button to proceed using the pre defined values with the risk of ending with a result that is out of range for the defined bed height END FILLING can also be used to abort the packing e Press PAUSE to temporarily pause the procedure In the next dialog press ABORT to quit packing or CONTINUE to proceed AxiChrom 300 1000 columns User Manual 28 9562 89 AD 127 5 Packing the column 5 4 Packing using the Master Wizard Step 10 Rinse the media valve Packing progress Rinse 10 15 ena Capto Q Rinse Adapter position 117 cm Keep the slurry tank valve open Start the pump at 503 Uh through the J Rinse hose further into the slurry tank When
69. ct The bed height is the same as the adapter position and should always be read from the level scale located on the adapter rod Note The level scale indicator differs between columns with different bed support material plastic or stainless steel Make sure that the correct level scale indicator is fitted If the value is incorrect change it by pressing MENU and then in the MAIN MENU Press BACK to reach the Verify adapter position dialog and see Section 3 7 1 Start Master and calibrate column on page 75 for more information about changing the bed height 2 When the values have been confirmed press CONTINUE to proceed to the next step 154 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes Step 5 Connect the pump Pump settings n m awam Capto Q To unpack the column you need to have an extemal pump connected and have a setup that allows both upwards and downwards flow during the unpacking procedure Please verify that you have a pump connected to mobile phase Make sure your pump is capable of providing the flow rates listed below Pump requirements Expansion phase 377 Uh Unpacking phase 151 h Connect a pump to the Bottom mobile phase valve Ensure that
70. cted to the fitting on the front of the trolley bar Disassembling the wheels on AxiChrom 800 and 1000 columns 1 56 Adjust all feet until they have the same distance between the floor and stand 95 135 mm AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 3 Wheel kits 2 Place the jack J under the frame at the reinforced pad towards the rear wheel and lift Lever the back of the column using the jack 3 Remove the rear wheels from the hole on the column stand 4 Lower the column again and remove the jack 5 Remove the second pair of rear wheels on the other side of the column by re peating steps 1 to 4 6 Place the jack at position I in the figure Use the jack to lever the front of the column so that the wheel kit can be removed from underneath the trolley bar G Lower the column front again and remove the jack The column should now stand on its front feet G gt G K AxiChrom 300 1000 columns User Manual 28 9562 89 AD 57 3 Getting started 3 3 Wheel kits 10 11 12 13 14 Fit the trolley bar wheels in the trolley bar storage positions as the figure shows Fit the rear wheels in the trolley bar storage positions F Attach the handle to the trolley bar Unscrew the screw E on one of the trolley bar sides and insert the bracket H between the trolley bar and the co
71. ctly placed to se cure the rear wheels 3 Lower the column by lifting the lever arm Ensure that the column stands firmly on the wheels before removing the lever 4 Fit the second pair of rear wheels on the other rear foot by repeating steps 1 to 3 above 50 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 3 Wheel kits 5 Attach the trolley bar to the front of the column stand Make sure that the pins underneath the stand F Wheel and trolley fittings on page 49 are properly fitted into the slots E of the trolley bar Trolley and trolley bar on page 49 before tightening the bolts B to the column stand 6 Place the trolley knob D Trolley and trolley bar on page 49 vertically under the adjusting screw C and adjust the height adjusting screw so it rests on the trolley knob 7 When the height adjusting screw is adjusted secure it with the safety pin S 8 Lift the front of the column by lowering the handle on the trolley Turn the trolley so it faces forward 9 Ensure that the column fully rests upon the wheels Screw the foot pads in completely The column can now be moved Disassembling the wheels on AxiChrom 300 600 columns 1 Adjust all feet until they have the same distance between the floor and stand 90 to 110 mm AxiChrom 300 1000 columns User Manual 28 9562 89 AD 51 3 Getting started 3 3 Wheel kits 2 Turn
72. cycle 1 60 cycle 2 30 cycle 2 Custom default settings 75 60 30 60 186 AxiChrom 300 1000 columns User Manual 28 9562 89 AD C Accessories and additional literature Appendix C Accessories and additional literature Tubings Tube S Cor 3 8 ID 9 5 TC25 ID 7 75 2000 mm 28 9377 58 Tube S Cor 3 8 ID 9 5 TC25 ID 7 75 5000 mm 28 9381 24 Tube S Cor 1 2 ID 12 7 TC25 ID 9 4 2000 mm 28 9381 25 Tube S Cor 1 2 ID 12 7 TC25 ID 9 4 5000 mm 28 9381 26 Tube U Cor 3 4 ID 19 1 TC25 ID 15 75 2000 mm 28 9381 27 Tube U Cor 3 4 ID 19 1 TC25 ID 15 75 5000 mm 28 9381 29 Tube U Cor 1 ID 25 4 TC50 ID 22 1 2000 mm 28 9381 36 Tube U Cor 1 ID 25 4 TC50 ID 22 1 5000 mm 28 9381 37 Tube U Cor 1 ID 38 1 TC50 ID 34 8 2000 mm 28 9381 38 Tube U Cor 1 ID 38 1 TC50 ID 34 8 5000 mm 28 9381 39 T junctions Article name Code no Tee 3 8 3xTC25 ID 7 75 28 9381 67 Tee 1 2 3xTC25 ID 9 4 28 9381 68 Tee 3 4 3xTC25 ID 15 75 28 9381 69 Tee 1 3xTC50 ID 22 1 28 9381 70 Tee 1 3xTC50 ID 34 8 28 9381 71 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 187 C Accessories and additional literature Reducers Article name Code no Reducer TC25 ID 7 75 TC25 ID 9 4 28 9381 73 Reducer TC25 ID 9 4 TC25 ID 15 75 28 9381 74 Reducer TC25 ID 15 75 TC50 ID 22 1 28 9381 75 Reducer
73. dard J T11363 2006 Requirements for Concentration Limits for Certain Hazardous Substances in Electronics AxiChrom 300 1000 columns User Manual 28 9562 89 AD 25 2 Specifications and characteristics 2 Specifications and characteristics 2 1 Principles of column function Intelligent packing Intelligent Packing uses interactive guides to ensure an optimized compression of the bed Intelligent Packing will pack the column with minimal manual intervention to ensure reproducible results Intelligent Packing is controlled from AxiChrom MasterMaster either as a stand alone unit or controlled in turn from an AKTAprocess instrument running UNICORN software Axial compression AxiChrom columns are packed using axial compression achieved by an electric motor driving the adapter downwards through the column tube until the required packing is achieved Filling the column To fill the column the adapter is raised to draw media slurry through the media valve inlet into the process chamber i e the space between the adapter bed support and bottom bed support The correct slurry volume to draw into the column is calculated using e the target bed height e the slurry concentration e a Packing Factor for optimal packing defined later e the volume added by the tube leading from the slurry tank to the column 26 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 2 Specifications and characteristics 2 1 Pri
74. e Target Packing Factor Filling speed cm h Packing speed cm h 124 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 4 Packing using the Master Wizard Note The recommended filling speed is 300 cm h for optimal mixing in the column during the filling step However if the slurry concentration is high gt 709 or if the slurry viscosity is high for example in a cold room lt 10 C the filling speed should be reduced to 200 cm h If the filling speed is too high cavitation may occur leading to formation of air bubbles inside the column The following settings are used for bed detection and may be changed for packing media not in the list on page 183 e Bed detection delay s e Bed detection level mbar Press the CONTINUE button to save the changed settings or press BACK to return without saving Step 7 Verify the settings Verify the settings The Summary dialog displays all selections made in the previous di alogs and also the calculated required slurry volume in liters Column 800 300 Media Capto Q Slurry concentration 65 Target bed height 10 0cm Max bed height limit 11 0cm Min bed height limit 90cm Required slurry T Packing factor 1 10 Filling speed 300 cmh Packing speed 60 cmh Press the CONTINUE button to confirm the settings or press BACK to return without saving AxiChrom 300 1000 columns User Manual 28 9562 89 AD 125 5 Packing the column 5 4 Packing usin
75. e During the priming procedure the adapter pushes the liquid out through the mobile phases at up to 300 cm h If a system is connected make sure that it can handle this flow and that the pressure drop in the system is not too high at that flow The flow deviation alarm and the air alarm in UNICORN or alarms of equal functionality in other systems must be disabled since they may be triggered by a velocity higher than that the system is designed for or by air bubbles flowing through the system Do not disable the pressure alarms on the system If an alarm is triggered on the system the adapter must be stopped immediately otherwise the adapter will continue to move and increase the pressure in the column up to 4 bar at which the AxiChrom Master pressure alarm goes off Step 3 enter pump capacity Enter the flow that vall be used for filling the column Max recommended liquid velocity for an unsupported screen is 400 cmh Max pump flow capacity 1800 I h Entered flow equals a flow velocityof 358 cm h 1 Enter the flow that will be used for filling the column by pressing Set The maxi mum recommended liquid velocity for an unsupported bed support is 400cm h Enter the value in liters hour I h the entered value will then automatically be converted to cm h 2 Press the CONTINUE button to proceed Step 4 check preconditions Check precondition boxes when the conditions have been fulfilled The priming can then be initiated by pres
76. e Intelligent Packing wizard and return to the MAIN MENU AxiChrom 300 1000 columns User Manual 28 9562 89 AD 131 5 Packing the column 5 4 Packing using the Master Wizard Bed height out of limit Packing progress 12 15 Warning The specified bed height range cannot be dpi patton 1218 ta met Press Abort to go to main menu Adapter speed 0 cmh 1 Consolidated bed detected at 12 6 cm Adapter position 12 6 cm Packing factor 1 00 Pressure 0 03 bar Adapter stopped Media valve closed ADO The specifications for bed height range and Packing Factor range can not be met That is the bed was detected but the specified bed height range is out of limits from the Packing Factor range optimal Packing Factor 0 03 Press Abort to end the Intelligent Packing wizard and return to the MAIN MENU Step 13 Monitor the compression Packing progress 13 15 Packing factor Bad height Consolidated bed detected st 110cm Adapter position 106 cem Packing factor 1 04 Distance to target packing factot 0 6 cm tortor 103 Time to min packing factor o19 Time 10 max packing factor O52 1 10 10 0 it 18 Stop adapter as close to target packing factor 1 10 as possible for optimal compression 90 Vem OESEL aa 132 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 4 Packing using the Master Wizard The compression of the bed has now started The accepted Packing Factors are shown on the left side o
77. e System will flow upwards and unpack the media through the Media valve to the slurry tank as the adapter moves downwards During the last push out step it is possible to change the adapter speed in Pump Increase_Adapter_Speed under Manual Instructions to speed up the unpacking procedure It is not possible to change adapter speed if the pressure is too high gt 1 bar or gt 0 5 bar depending on media When the pressure comes below the limit for the media it is possible to change the adapter speed again The maximum unpacking adapter speed is 240 cm h 3 When the adapter is at the bottom position a flush of remaining media in the column is initiated When the bed support is free from media click Continue Result The Media valve closes and a rinse of the valve and slurry tube is started 4 When the slurry tube is free from slurry click Continue 5 Close the slurry tank valve and click Continue Unpacking acrylic column tubes using Master wizard The unpacking procedure described in the Master wizard guides the operator through a number of steps to unpack the column with manual valves and a standalone pump Follow the instructions in the Unpacking wizard found in the MAIN MENU in AxiChrom Master For tubing connections see Manual configuration on page 69 Note This wizard changes depending on the media type and media chosen The method described below is a general method for the unpacking of Capto and Fast Flow media A different un
78. e discs can be or dered as accessories from GE Healthcare Ensure that any liquid that is ejected by the over pressure protection equipment is con nected to a waste outlet 68 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 6 Column and system setup Manual configuration The following manual setup makes it possible to perform all processes available in the AxiChrom Master PRIMING INTELLIGENT PACKING and UNPACKING The setup com prises two way valves and shut off valves to utilize different flow paths for the different procedures To waste o FESI e Column Slurry inlet Rinse inlet To waste To waste Symbol Function Pressure relief valve rupture disc or similar T piece 2 way membrane valve shutoff valve 2 way membrane valve shutoff valve with bleed can be replaced with gt lt two 2 way membrane valves and a T piece AxiChrom 300 1000 columns User Manual 28 9562 89 AD 69 3 Getting started 3 6 Column and system setup Possible flowpaths for different column operations Process Flow path alternatives from gt to Pump gt bottom mobile phase gt waste valve 5 Priming Column gt bottom mobile phase gt waste valve 6 Packing Column gt bottom mobile phase gt waste valve 6 Rinse pump gt valve 2 gt valve 4 gt rinse gt slurry tank Unpacking pump g
79. e used The valve holder is clamped on the front stand tube with a U rod and holds the components with 3 pronged clamps One valve holder carries two clamps Different sizes of U rod are used for AxiChrom 300 600 columns and AxiChrom 800 1000 columns respectively AxiChrom 300 1000 columns User Manual 28 9562 89 AD 29 30 2 Specifications and characteristics 2 2 Overview of the column and parts Valve holder accessory inset and mounted with valves and sensors on front stand tube AxiChrom 300 1000 columns User Manual 28 9562 89 AD 2 Specifications and characteristics 2 2 Overview of the column and parts AxiChrom Master AxiChrom Master is a self contained operator console featuring interactive guides for work procedures such as packing unpacking and maintenance The user interface is a touch screen panel The interactive firmware guides the user and reduces the risk of making mistakes The operator has control over the workflow and can use manual control for adapter move ment and open or close the Media valve One AxiChrom Master unit can be used to control up to ten columns one at a time Note Always use the same AxiChrom Master to control a given column The motor will be recalibrated automatically if you change to a different Master AxiChrom Master specifications Parameter Value Weight Approximately 73 kg 161 Ibs Length x width x height 670 x 590 x 1090 mm Material Stainless
80. eiet Derion vae inlet Mole phase m i aaa fat fa sytem COLUNN TOP potion a connected lo the adist on the axe of the OP vabe Connect the AsiChrom Master pressure tarsmitier PIS_113 to system INLET CIPI Correct the column moble phase top to the AxiChom Master protse tuarusration PIS_119 Correct the coke mabie phase bettom to the system COLUMN BOTTOM valve Correct the column rinse irist to system INLET CIP2 Correct the shar tark to the column modis rist AxiChrom lt Back This page provides a summary of the options selected in the previous wizard pages An estimated buffer consumption is given and how to connect the column to the system is described 5 Click on Finish The unpacking summary can be viewed under Notes Method Notes in the Method Editor module of UNICORN 6 Choose an appropriate result file folder result name and save the method Creating a custom unpacking method Unpacking methods may also be developed for custom media using the Method Wizard The wizard facilitates this by providing an extended range of variables that may be modified to optimize the method 1 Start the Method Wizard in the Method Editor window and choose a system in the pop up window 2 Select the appropriate AxiChrom column 3 Select All or Other media type and then select Custom medium 4 Click Next to continue in the wizard For custom media it is possible to choose from two unpacking modes regular or ad vanced
81. entered on the dome of the disc then place the rupture disc into the ferrules with the flow arrow on the rupture disc tag pointing in the same direction as the desired vented flow after rupture NOTICE gt Always handle rupture discs with extreme caution 5 Install the clamp around the ferrules so that the gap between the two clamp halves is centered on the rupture disc tag 6 Hand tighten the wing nut until the clamp is fully engaged with the ferrules and the rupture disc is securely in position The nut will turn freely until it becomes snug with the clamp Tighten another 1 to 1 25 turns 7 Perform column and system setup according to the relevant instructions in Section 3 6 Column and system setup on page 68 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 67 3 Getting started 3 5 Rupture discs CAUTION Double check the orientation of the rupture disc Verify that the flow arrows on the disc tags are pointed in the same direction as the desired vented flow after rupture 3 6 Column and system setup In this section This section describes how to connect the AxiChrom column to tanks pumps and for example an AKTAprocess system for priming packing and unpacking sequences in the AxiChrom Master WARNING A pressure relief valve rupture disc or a similar over pressure pro tector must be installed at both of the mobile phases before the column is used Pressure relief valves and ruptur
82. erage particle size of the media This ensures that all particles are larger than the holes in the bed support Using a bed support with a larger porosity results in particles passing or getting caught in the screen leading to poor bed efficiency and for a clogged screen high pressure Preparation of the slurry Note Prepare the slurry at least 2 hours before usage in order for it to properly degas after shaking The slurry can be prepared up to 1 day ahead Tip In the case of packing short beds in the range of 10 to 15 cm it is often suitable to first reduce the slurry concentration to about 30 to 35 Further it is rec ommended to increase the Packing Factor from the set value in the Master by 0 02 or 0 03 2 to 3 Using either or both of these modifications often leads to improved results Procedure 1 Calculate the amount of media using the formula media volume target bed height x PF x column cross section 2 Decant the supernatant and replace with recommended packing buffer accord ing to the tables on page 183 until a suitable slurry concentration is reached Short column tube at least 30 for 100 mm bed height and 65 for 300 mm bed height Long column tube at least 30 for 100 mm bed height and 74 for 500 mm bed height Note Minimum concentration bed height x PF or CF max fill height 5 Determine the slurry concentration see Section 4 3 Determining the slurry con centration on page 91 AxiChrom 300
83. ess steel bed supports Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids 180 AxiChrom 300 1000 columns User Manual 28 9562 89 AD A Parts list and diagrams A 5 Media valve and tubes A 5 Media valve and tubes Columns with stainless steel tubes and 800 and 1000 columns with PP tubes Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids AxiChrom 300 1000 columns User Manual 28 9562 89 AD 181 A Parts list and diagrams A 5 Media valve and tubes 300 600 columns with PP tubes Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids aa gt SH 2 DD S NE u N B A 182 AxiChrom 300 1000 columns User Manual 28 9562 89 AD B Packing buffer information Appendix B Packing buffer information Packing buffers and packing factors Packing values Note The Packing Factor range used in the verified packing procedure in the Master or UNICORN is PF 0 03 The range has been verified to give stable efficient packed beds Media not listed here may be available in UNICORN or AxiChrom Master Packing
84. f the animation and the bed height limits are shown on the right side of the animation The area in which these two criteria overlap shown as green in the animation is where an approved packed bed can be achieved The adapter shown as the black horizontal line is moving down in the animation closing in on the green ap proved range of Packing Factor and bed height Note If the picture for some reason does not resemble this example above press Reset to refresh the picture Step 14 Finish the packing Packing factor Bed height ane POCENIE ji us Consolidated bed detected at 11 0 cm Adapter position 10 3 cm Packing factor 1 08 Distance to target packing factor 0 2 cm Calculated packing pressure 0 1 bar Stop adapter as close to target packing factor 1 10 as possible for optimal compression 90 End packing When the adapter is within the limits within the green area the new button End packing will appear Press End packing to stop the adapter as close to the target Packing Factor the static black line in the green area as possible for optimal compression AxiChrom 300 1000 columns User Manual 28 9562 89 AD 133 5 Packing the column 5 4 Packing using the Master Wizard If the adapter is not stopped If the adapter is not stopped a dialog like the following is displayed Packing progress Warning Packing factor Bed height g no The adapter was stopped due to overcompression 66 HELP of the user manual for
85. further instructions 10 100 Adapter position 9 5 cm Packing factor 1 16 Calculated packing pressure 0 1 bar The adapter stops automatically 15 seconds after the optimal Packing Factor PF 0 03 is reached or e atthe low bed height limit In the first case the bed has been over compressed Press Confirm to proceed Press Abort to end the wizard and return to the MAIN MENU See Method 2 on page 139 for help about over compression Step 15 Review the process summary Packing completed 5 15 ean Capto Q The packing of Capto Q in column 800 300 is complete Bed height 10 0 cm target 10 0 Column volume 50 37L Pack factor 1 10 target 1 10 Consolidated bed height 110cm This dialog summarizes the packing process and displays these values 134 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 4 Packing using the Master Wizard Resulting bed height and target bed height Column volume The column volume is calculated using several decimal place s precision in both the value of Pi and adapter position This may lead to a discrepancy in the displayed volume compared to that which can be calculated from the figures shown Resulting Packing Factor and target Packing Factor Height where the consolidated bed was detected Press the MAIN MENU button to leave the AxiChrom Master Intelligent Packing wizard 5 5 Creating custom packing methods in the Master Methodology for choos
86. g the Master Wizard Step 8 Check the preconditions for packing 126 Check the preconditions for packing Preconditions 8 15 aoa Capto Q Before you can start packing the following preconditions need to be fulfilled The hoses system column and media valve are pnmed The Rinse inlet is connected to a system or a pump and is closed Both top and bottom mobile phase valves are closed A Slurry tank is connected to the Slurry Inlet The slurry tank valve is open Make sure the media is homogeneously suspended Make sure the slurry volume is at least 86 L SS AINSI SA The tubing system column and Media valve are primed The Rinse inlet is connected to a pump and is closed Both the Top and Bottom mobile phase valves are closed A slurry tank is connected to the Slurry inlet on the column The valve on the slurry tank is opened The media is kept homogeneously suspended throughout the filling process The slurry volume is at least the specified value Check the boxes and press START to initiate the packing process AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 4 Packing using the Master Wizard Step 9 Fill the column Packing proaress Filling bie Capio Q 251 romaning 7 Adapter commanded posdion 18 1 cm an Adapter position 38 cm Adapter speed 300 cmh Filling in progress The Filing vall stop automatically when finished END FILLING wall
87. ge the adapter speed The speed can be changed again when the adapter starts to move Press PAUSE to stop the movement temporarily or to terminate the UNPACKING wizard Step 13 Stop the pump progress Adapter is at the bottom position 13 16 Adapter Is at the bottom position Stop the pump and press CONFIRM Adapter position 10 cm Pressure 0 04 bar Adapter stopped Media valve open CONFIRM 1 Stop the pump when the adapter is in its bottom position If the pump is not stopped the slurry will be diluted 2 Press CONFIRM to acknowledge Step 14 Flush out the remaining media npacking progress Flush the remaining media out 14 16 enue Q Sepharose High Performance Flush the remaining media out Adapter position 10 0 cm To flush emanning media out start the pump at upfiow from the bottom mobile phase See Help for further instructions Press CONFIRM when the column is empty and the pump has been stopped Pressure 0 04 bar Adapter stopped Media valve closed CONFIRM If there is media left in the column when the adapter has come to the bottom position itcan be flushed out Use longer flushing times for more efficient media removal Collect the flushing liquid in a separate vessel to avoid diluting the previously unpacked slurry Press SKIP if you do not want to flush out the remaining media Follow the steps below to flush out the remaining media AxiChrom 300 1000 columns User Manual 28 9562 89 A
88. has fallen to the bottom of the column the Media valve opens and the slurry can be pushed out from the column by having a low upwards flow into the column and at the same time lower the adapter to the bottom position Unpacking is performed either using UNICORN on an AKTAprocess system or from the AxiChrom Master directly The workflows for unpacking acrylic and stainless steel column tubes are described in Section 7 3 3 Unpacking acrylic column tubes on page 150 and Section 7 3 4 Unpacking stainless steel column tubes on page 162 respectively 7 3 2 Creating a UNICORN method The Method Wizard in UNICORN simplifies creation of an Unpacking method The methods require an initial selection of a listed medium Thereafter the user makes appro priate selections to complete the creation of the method This section describes the creation of a UNICORN unpacking method If a custom medium is selected in the Method Wizard media list a wider range of variables are allowed to be defined to optimize the method for two different unpacking procedures that can be used see Creating a packing method based on custom media on page 115 Procedure 1 Start the Method Wizard in the Method Editor window and choose a system in the pop up window 146 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 3 Unpacking the column 7 3 2 Creating a UNICORN method 2 Select Column Unpacking Mulliud WE ur orale ATAY ub ans act GDU bonlo
89. he pressure on sensor PRESS_119 Set the vertical scale on the pressure graph for PRESS_119 to minimum 0 1 bar maximum 0 5 bar and make sure that the horizontal axis displays time Automatic bed detection is triggered by a drop in pressure by 0 01 bar for a duration of at least 2 seconds Manual bed detection is indicated by a corresponding drop in the pressure graph AxiChrom 300 1000 columns User Manual 28 9562 89 AD 137 5 Packing the column 5 5 Creating custom packing methods in the Master 5 5 1 Packing stainless steel column tubes Figure 5 1 The blue trace shows the pressure recorded at PRESS_119 during column packing Bed detection is indicated by a transient pressure drop arrow Packing the column using the Master wizard For the general packing procedure using the Master wizard see Section 5 4 Packing using the Master Wizard on page 120 With stainless steel column tubes it is not possible to see when the adapter is approach ing the consolidated bed surface step 12 For manual bed detection verify that the adapter has reached the consolidated bed by watching the pressure indication in the Master dialog Automatic bed detection is triggered by a drop in pressure by 0 01 bar for a duration of at least 2 seconds Manual bed detection is indicated by a corresponding drop in the pressure graph 5 5 2 Post packing modifications Method 1 If there is reason to believe that the bed needs further compression f
90. he column 4 4 3 Creating a UNICORN method 3 96 pen lt oe k ArT A Pid Setting Prime media valve and slurry hose Click Next The wizard page Priming Parameters opens Select the appropriate inlet Description Leave checked to prime the media valve during the priming process Inlet for mobile phase Select the appropriate inlet AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 4 Priming the column 4 4 3 Creating a UNICORN method 4 Click Next to display the wizard page Priming Summary MLI Hurd lug sy aik ANIA piueese Ater Kuali Corrected to the outlet on the ade of the CIP valve Connect a memtzare vatve to system INLET OPI Connect the AudCheom Master pressure trancmates PIS_119 tote menteare vave Connect the colma mebile phase top to the AxiChiom Mane preceuse vanamines PIS_119 Connect the column mobile phase bottom to the spitem COLUMN BOTTOM vahe Connect the cokane rinse irket to system INLET CIP2 Connect the shiny tark to the column shary irist AxiChrom This page provides a summary of the options selected in the previous wizard pages The summary page also gives information on buffer volumes and instruc tion for how to connect the column to the KTAprocess system 5 Click Finish The column packing summary can then be viewed under View Run Setup Notes Method Notes in the Method Editor module of UNIC
91. he column is primed e The flowpath to the column is primed e Aslurry tank is connected to the Slurry inlet Check the boxes in the dialog and press START to proceed Step 7 Start the pump upflow npacking progress Start the pump 7 16 aoa Capto Q Start the pump upflow Start an upwards flow through the column at 377 Ih Adapter position 10 0 cm The pump has reached a flow of at J least 377 ih Press CONFIRM to start the adapter movement Pressure 0 03 bar Adapter stopped Media valve closed CONFIRM Start the pump upflow through the column at the flow stated in the dialog When the pump has reached a flow of at least the stated value check the box and press CONFIRM to start the unpacking AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes Step 8 Wait for bed expansion npacking progress Expanding bed 8 16 amwa Capio Q 217 romaning Adapter commanded postion 14 0 om LELEL Adapter position 11 7 cm Adapter speed 61 cmh Expanding the bed Naroase the PUN IOW selects before increasing the adapter speed The pump Roxy Vooy SOUT esis De Nigher han the adapter speed Adapter speed Pressure 0 29 bar adjustment Adapter moving Media valve closed Defaut speed 60 cmh PAUSE e Increase the pump flow velocity before increasing the adapter speed The pump flow velocity should always be highe
92. high degree of resistance to corrosion they are susceptible The degree of susceptibility is largely due to conditions of use Low pH in combination with NaCl high temperature and surface damage all increase risk of corro sion If a stainless steel surface is damaged scratches impact marks etc is has to be cleaned and passivated prior to taken into operation Your GE Healthcare representative can provide guidance on suitable cleaning and passivation procedures if required To clean the column from chloride ions it is recommended that a water rinse of at least five column volumes is used Water is the preferred rinse solution due to the solubility properties of sodium chloride in water compared to other possible rinsing solutions for example ethanol solutions Columns should be stored with solutions free from chloride ions AxiChrom 300 1000 columns User Manual 28 9562 89 AD 39 3 Getting started 3 Getting started 3 1 Unpacking the crates and cleaning a new column Unpacking the crates The following manuals contain information about how to unpack a new column e Read Me First contains specific instructions on how to unpack the column and Axi Chrom Master from their crates The instructions are attached to the outside of the packing crates upon delivery Refer to those instructions when unpacking the products from their crates e AxiChrom columns Site Preparation Guide provides guidelines and requirements re garding how to place
93. imit and to the calculated bed detection are displayed in the right frame The current position and the speed of the moving adapter are displayed in the left frame Bed detection Waiting for detection Packing progress 12 15 awam 1 Orisa Bed detection window Pa Waiting for detection a oo oo Adapter position 13 5 cm Pressure 0 24 bar Time to calculated bed detection 2 27 Pre Adi Me AxiChrom 300 1000 columns User Manual 28 9562 89 AD 129 5 Packing the column 5 4 Packing using the Master Wizard This dialog will automatically open 3 to 6 minutes before the calculated bed detection occurs The time depends on the media with longer time for High Performance media When the adapter reaches the bed the user has two options e Let the AxiChrom Master detect the contact between the adapter and the gel bed e Manually detect that is visually verify that the adapter has contacted the bed by pressing Detect to proceed to the next step in the packing procedure Note Manual detection should only be used if the user is certain that the adapter has indeed come into contact with the consolidated bed but no automatic detection was executed The Axichrom wizard will apply the compression specified for the selected medium starting from the point of execution of the selected option Bed is detected Bed is detected Manual detection Adapter position 10 8 cm Pressure 0 03 bar Consolidated bed detected at 11 0 cm Pack
94. ing packing parameters Using the Intelligent Packing wizard the factors that can be varied to optimize the packing results for a particular media are mainly Packing Factor and packing speed The packing buffer may sometimes also play a significant role For more information about packing factors and buffers see Section 4 1 Packing Factor and Compression Factor on page 89 and Appendix B Packing buffer information on page 183 Investigate what parameters are suitable for the media Check literature with the manufacturer or data from smaller scale columns It is very likely that the Packing Factor packing buffer or the compression can be found or at least estimated In the standard Packing wizard see Section 5 3 Packing using a UNICORN method on page 117 enter the known Packing Factor or a low value where it is known that there is no damage incurred to the media The packing speed is mainly dictated by the size of the media particles and poten tially also their density and rigidity In general smaller and less rigid media in the range from 15 to 40 um in diameter are suitable to pack at 30 cm h while larger particles around 100 um are best packed at 60 cm h Very large rigid and dense particles can be packed at 90 cm h even though 60 cm h often works well AxiChrom 300 1000 columns User Manual 28 9562 89 AD 135 5 Packing the column 5 5 Creating custom packing methods in the Master Changing parameter settings If the paramete
95. ing factor 1 03 i the detection is not confirmed the process will stop in 13 seconds There will be a 2 second delay before the bed detection dialog appears A consolidated bed has been detected automatically or manually e Press Confirm to acknowledge the detection and continue to the next step If not confirmed the process will stop automatically in 30 seconds e Press Ignore if the adapter is not in full contact with the bed The bed search will continue 130 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 4 Packing using the Master Wizard Unconfirmed detection Packing progress 12 15 100000sara WE DEAE Sophene F Bed detection window Bed is detected Manual detection Adapter position 11 0 cm Pressure 0 11 bar Consolidated bed detected at 0 0 cm Packing factor 1 05 The adapter has stopped due to unconfirmed detection F O ADO The adapter has stopped automatically since the bed detection not was confirmed manually The actual Packing Factor is displayed No bed was detected Packing progress 12 15 Warning The adapter has stopped at the lowbed height limit No bed was detected raai sone bub Press Abort to go to main menu Pressure 0 03 bar Adapter stopped Media valve closed ADOT The adapter has stopped at the low bed height limit without finding any bed This may happen if the wrong slurry concentration has been set Press Abort to end th
96. it is recom mended to use the default Packing Factor Unchecking this option allows the Packing Factor to be changed Note Over compression during packing can damage the medium The default Packing Factor for each medium has been tested and verified by GE Healthcare AxiChrom 300 1000 columns User Manual 28 9562 89 AD 113 5 Packing the column 5 2 Creating a UNICORN method 5 Click Next to display the Slurry and Packing Test Parameters wizard page Make the appropriate selections asinu Wiad up syal U lA piut sss Aaiun Lualiul Shay and Packing Test Parameters Estimated dury concenti aion 36 60 X a measured n demonized water 7 Enter the meansed conceriation in the start protocol Equiteston vokume 0 1 20 CV 10 E Use dota kgad velociy daing equlibeation gud velocity dunng eqaita son 1mh E Use delat kadd velocky daing paching test Laud velouty dung packing lest Dem E Packing test upiiow 2 Packing test downtion Inlet for sarge ede AxiChrom Back ja Nety CoR Cote Setting Description Slurry concentration Enter the estimated slurry concentration The measured concentration is entered later in the start protocol Equilibration volume Enter an equilibration volume measured in CV within the designated range Use default liquid velocity during equilibration pack ing test Leave checked to use the suggested default liquid velocity Packing tes
97. ith any bed support material Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids 176 AxiChrom 300 1000 columns User Manual 28 9562 89 AD A Parts list and diagrams A 3 Columns with plastic bed supports A 3 Columns with plastic bed supports Note The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids a DOO N aA a m a ft amp Ol oO AxiChrom 300 1000 columns User Manual 28 9562 89 AD 177 A Parts list and diagrams A 3 Columns with plastic bed supports Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids 178 AxiChrom 300 1000 columns User Manual 28 9562 89 AD A Parts list and diagrams A 4 Columns with stainless steel bed supports A 4 Columns with stainless steel bed supports Note e The numbers correspond to position numbers in Material Conformity and Spare Parts e Grey circles indicate parts which retain pressure or are in contact with process fluids AxiChrom 300 1000 columns User Manual 28 9562 89 AD 179 A Parts list and diagrams A 4 Columns with stainl
98. k the boxes and press CONFIRM to proceed Step 10 Stop the pump Inpacking progress Collapsed bed 10 16 aoa Capto Q 1 The bed has collapsed Adapter position 14 0 cm The bed is now collapsed Stop the pump J Pressure 0 03 bar Adapter topped Media valve closed Stop the pump Check the box and press CONFIRM to continue to the next dialog Step 11 Empty the column Unpacking progress Empty the calumn 11 16 awam Capto Q Adapter position 14 0 cm the column Open the slurry tank valve Close the top mobile phase Fi Start a flow upwards through the J column at 151 Uh 1 158 When pump is started press CONFIRM Pressure 0 03 bar Adapter copped Media valve open CONFIRM Open the slurry tank valve AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes Z Close the Top mobile phase inlet outlet 3 Start an upwards flow through the column at the flow stated in the dialog 4 When this flow has been reached check the boxes and press CONFIRM to proceed Step 12 Wait for unpacking to the slurry tank 1235 remainng Adapter commanded postion 1 0 cm Adapter position 13 6 cm Adaplee speed EO cmh Unpacking to the slurry tank Reduce the adapter speed if the adapater contacts the bed Adapter speed Pressure 0 03 bar adjustment Adapter moving Media valve opon Default speed 60 cmh e
99. ket between the column stand and the trolley bar as in H Steer the wheel kit with the trolley handle and lift up until part C reaches the lower surface of the column stand Make sure that the pin underneath the stand fits inside the slot D of the trolley bar then tighten the screw E in the image in step 1 Make sure that both screws E are in the correct position Remove the handle from the trolley bar The handle is only a help when putting the trolley bar into position and should not be used when moving the column AxiChrom 300 1000 columns User Manual 28 9562 89 AD 55 3 Getting started 3 3 Wheel kits 10 it Lower the column front again and remove the jack The column should now stand on the front wheels of the trolley bar and the front feet should not touch the ground Both wheels should be locked facing the direction of the frame legs before continuing to the next step Place the jack J under the frame at the reinforced pad towards the rear wheel and lift Fit the rear wheels in the hole A in the column stand see illustration on page 54 Ensure that the washer and safety pin are correctly placed to secure the rear wheels Lower the column again and remove the jack Make sure that the front feet do not touch the ground Fit the second pair of rear wheels on the other side of the column by repeating steps 7 to 9 Ensure that the column rests fully on the wheels The column can now be moved using a MasterMover conne
100. les that may be modified to optimize the method For example it is possible to e Select filling speed in a defined range AxiChrom 300 1000 columns User Manual 28 9562 89 AD 115 5 Packing the column 5 2 Creating a UNICORN method Pack according to a desired Packing Factor The custom method utilizes the same sequence for packing as the regular method The adapter must be stopped in the overlap of defined min max bed height interval and the Packing Factor range Edit detection limits Select flow conditioning with associated variables Flow conditioning is always per formed down flow unless the flow direction is changed in the variables page Specify desired adapter velocity during packing within defined range Select sample volume within defined range Procedure Start the Method Wizard in the Method Editor window and choose a system in the pop up window 2 Select whether to create a method for Column Packing Packing Test or the two combined 5 Select the appropriate AxiChrom column 4 Select All or Other media type and then select Custom under Medium o A EAE AE AA Inteligent Packing Cokann Packing vath Packing Test a Cmn AvdChrom 450 300 v Coumn maesi Acn c v Shon medum ype Other a Modam Custom AxiChrom Nest gt cma Hop Sedma 5 Click Next to continue in the Method Wizard Note Over compression during packing can damage the medium The default 116 Packing Factor for each medi
101. ls All columns can be ordered with wheels as accessories Columns without wheels must be moved with a forklift pallet lift or lifted by a hoist or crane attached to the adapter rods A forklift or pallet lift with a distance of about 670 mm between the outer edges of the forks is recommended WARNING Ensure that the forklift has capacity to safely lift the column weight The column weight is shown on the rating plate WARNING When lifting with forklifts pallet lifters or similar devices the column must be lifted from the long sides Do not try to lift the column from the back the short side WARNING Ensure that the column s center of gravity is well balanced over the lifting forks otherwise the column may tip off the forklift NOTICE When moving the column using a forklift or pallet lifter make sure that the media valve under the bottom of the column is not dam aged Close the media valve so that it protrudes less NOTICE If the column is to be moved when it is filled plug the top and bot tom mobile phase inlets with tri clamp blind caps diaphragms or other suitable valves Oo O gt gt gt AxiChrom 300 1000 columns User Manual 28 9562 89 AD 45 3 Getting started 3 2 Placing the column and Master a 46 Forklift placing Always place the forks of the forklift or pallet lifter under the bottom of the column Make sure that the forks are properly placed in relatio
102. lume V Maximum column bed volume Design pressure PS Design pressure for the column Test pressure PT Pressure at which the column has been tested in manufacturing Date Date of test Maximum filling mass Maximum weight of column contents media and eluent PED fluid group cat Pressure Equipment Directive fluid group category AxiChrom 300 1000 columns User Manual 28 9562 89 AD 23 1 Introduction 1 10 Labels AxiChrom Master label The illustration below shows an example of an AxiChrom Master label AxiChrom Master 300 1000 Code Number 28803963 Enclosure Type 1 Serial Number Largest Motor Current FLA 5 8 A Manufacturing Year Max Current FLA 6A Supply Voltage Range CE 380 400V 3 Pneumatic Supply 5 5 7 bar g Max Voltage NA 480Y 277 V AC Short Circuit Rating 5kA Frequency 50 60 Hz Protection Class IP56 Max Power Consumption 2400 VA LAcc to NEMA 4X Mfn Site UM1 GE Healthcare Bio Sciences AB X Field Wiring Bj rkgatan 30 Diagram S 751 84 Uppsala i 28921816 Sweden Label text Meaning Code Number Product code number for AxiChrom Master Serial Number Serial number for the individual AxiChrom Master Manufacturing Year Year when the AxiChrom Master was manufactured Supply Voltage Range CE Permissible supply voltage range Europe Max Voltage NA Maximum permissible supply voltage
103. lumn The unpacking of the media from the column is performed in three steps e expansion e collapse e slurry push out The expansion is performed by lifting the bed by moving the adapter up at the same time as flow comes from the bottom bed support When the bed has expanded the flow is reversed to collapse the bed by pushing it down When the collapsed bed has fallen to the bottom of the column the Media valve opens and the slurry can be pushed out from the column by having a low upwards flow into the column and at the same time lower the adapter to the bottom position AxiChrom 300 1000 columns User Manual 28 9562 89 AD 27 2 Specifications and characteristics 2 2 Overview of the column and parts 2 2 Overview of the column and parts Part Function Part Function 1 Top unit 8 Bottom bed support and distribu tor plate 2 Worm gear and bellows 9 Column stand 3 Servo motor 10 Process chamber 4 Top mobile phase inlet outlet 11 Column tube 5 Adapter 12 Slurry inlet 6 Adapter bed support and distribu 13 Bottom mobile phase inlet outlet tor plate 7 Media valve assembly 14 Rinse inlet 28 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 2 Specifications and characteristics 2 2 Overview of the column and parts For information on column weights and dimensions see Appendix E Colu
104. lumn and Master setup Add or edit a column Size Diameter I Max bed height Bed support Steel Plastic The name cannot contain more than 30 letters The dialog is reached after pressing Add in the previous dialog Settings for a column can be created here The column diameter and maximum bed height can be found on the column label It is located on the lower front part of the frame on the column Select the correct column diameter and maximum bed height in the dialog by using the arrow buttons The chosen column will be highlighted Select the type of bed support fitted in the column to be used The options are Steel and Plastic Press Set to open the on screen keyboard where a unique column name can be written This name can contain a maximum of 30 characters Note It is recommended that a label with the column name that was given by the operator in the AxiChrom Master is placed on the column The Serial number is a good unique name Press SAVE to save the chosen settings for the column and to continue Press BACK to return to the Select column dialog without saving Editing the column name is possible even when the column has been saved 80 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 7 Using the Master 3 7 3 Column and Master setup Motor calibration alibration Please wait while the AxiChrom Master calibrates the motor Seeedeneeneee Calibration completed successfully
105. lumn stand Use the bracket to carefully lower the trolley bar down until the trolley bar rests upon the column stand Repeat step 10 to release the other side of the trolley bar Use the handle to retract the trolley bar from the column stand Ensure that the column stands firmly on all feet Ensure that the column is level see Placing the column on page 42 58 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 4 Connections Connections on Master and column 3 Getting started 3 4 Connections The illustrations below show the AxiChrom Master connector panel and the connection points on a column Part Description Description 1 Pneumatic air supply Motor feedback 2 Power switch Pressure sensor connector 3 Power supply cable Profibus signal cable connector 4 Motor power cable Two air outlet connectors AxiChrom 300 1000 columns User Manual 28 9562 89 AD 59 3 Getting started 3 4 Connections Part Description S 9 Slurry inlet 10 Bottom mobile phase inlet outlet 11 Rinse inlet 12 Top mobile phase inlet outlet 13 Motor power and motor feedback aT it al 14 Pressure sensor PIS_119 mounted on hose to system 15 Pneumatic inlets with 2 connectors Connect column Master and external s
106. ly depressurized before disconnecting any tubing or removing compo nents that are part of the flow path WARNING It is essential that access to EMERGENCY STOP buttons master power switch system power switch and power cable connections are kept free Do not block any of these items with other equipment or position the column so that they become unreachable WARNING A disconnecting system power switch must be installed such as a branch circuit protection and disconnect switch installed at your facility The system power switch should be installed at the wall to cut the power to AxiChrom Master Always use this power switch located on the back on the AxiChrom Master unit and the system power switch for safe disconnection of AxiChrom Master 1 10 Labels This section describes the identification labels safety labels and labels concerning haz ardous substances that are attached to the AxiChrom columns and AxiChrom Master 22 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1 Introduction 1 10 Labels Column label Label placement Example label Tr Tay ia Label text Meaning Dimension Dimensions of the column Serial number Serial number for the individual column Year of manufacturing Year when the column was manufactured Tare mass Weight of the empty column Operating temperature TS Operating temperature range Max column vo
107. m the connector of AxiChrom Master unit and replace the protective cap 7 Power on the AKTAprocess unit and log in 3 7 Using the Master 3 7 1 Start Master and calibrate column Description Every time the AxiChrom Master is started the column list is shown When a column has been chosen the Calibration page is displayed and the driver will automatically be cal ibrated to the column motor After calibration the Verify adapter position page is dis played Enter the adapter position as read on the column scale Firmware installation AxiChrom Master comes fully installed with the firmware needed To support new GE Healthcare media for Intelligent Packing or improved functionality including new column models AxiChrom columns ordered as separate units after AxiChrom Master delivery may have later firmware versions enclosed including compatibility information and in stallation instructions Please contact your GE Healthcare representative for further in formation AxiChrom 300 1000 columns User Manual 28 9562 89 AD 75 3 Getting started 3 7 Using the Master 3 7 2 User interface 3 7 2 User interface Master wizards The MAIN MENU is the starting point for wizards for PRIMING INTELLIGENT PACKING UNPACKING MANUAL CONTROL and MAINTENANCE Buttons and explanations Button General interface functions MAIN MENU Closes the ongoing operation and opens the MAIN MENU dialog or MENU The button is active when it is grey
108. mes 4 Pause the flow Use the membrane valves to seal the column mobile phases and incubate for 2 hours at 1 bar 5 Carefully release the pressure on the membrane valves 6 Flush the column with at least five column volumes of deionized water or packing buffer 7 Pack the column according to the AxiChrom Master or a UNICORN packing method 4 4 3 Creating a UNICORN method 94 The Method Wizard in UNICORN simplifies creation of a Priming method The methods require an initial selection of a column dimension Thereafter the user makes appropriate selections to complete the creation of the method This section describes the creation of a UNICORN Priming method Procedure il Start the Method Wizard in the Method Editor window and choose the system in the pop up window AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 4 Priming the column 4 4 3 Creating a UNICORN method 2 Select Priming AxiChrom nee Setting Description Column Select AxiChrom xxx yyy xxx column diameter in mil limeter yyy max bed height for short and long column tube 300 or 500 mm respectively The column size can be found on a label located on the lower front part of the frame of the column tube Column Select the material of the column tube material AxiChrom 300 1000 columns User Manual 28 9562 89 AD 95 4 Preparations before packing the column 4 4 Priming t
109. mitter PIS_119 to INLET CIP1 Connect a pressure relief valve or similar to PIS_119 Connect a tube of suitable length and inner diameter from the column Top mobile phase to the pressure relief valve on AxiChrom Master pressure trans mitter PIS_119 For tube diameter recommendations see Recommended mobile phase tubing inner diameters on page 62 Connect a tube of suitable length and inner diameter from CIP2 on the process system CIP Column Pack valve block to the Rinse inlet on the column Connect a tube of suitable length and inner diameter from Bottom mobile phase on the column to a pressure relief valve rupture disc or similar and then to Column1 bottom valve on the process system Connect the profibus cable from the Profibus connection situated under the electrical cabinet of the AKTAprocess system to the Profibus connection on the AxiChrom Master see Connecting AxiChrom Master to AKTAprocess via Profibus on page 73 Connecting AxiChrom Master to AKTAprocess via Profibus Connect the Profibus cable 1 Power down the AKTAprocess and Master units AxiChrom 300 1000 columns User Manual 28 9562 89 AD 73 3 Getting started 3 6 Column and system setup CAUTION Make sure that the AKTAprocess unit is powered off before the following step is performed Remove the Profibus termination plug from the AKTAprocess bulkhead connec tor Connect the Profibus cable to the AKTAprocess bulkhead connector Make sure
110. mn Due to the weight of the column tube assembly it is not advisable to try to stop the momen tum of the assembly by hand when it is set in motion WARNING There must not be any residues of harmful substances left in the column during maintenance Make sure that the column is properly cleaned before maintenance and that cleaning is documented in the Decontamination Report NOTICE Lifting the top unit with incorrectly placed lock blocks may damage the column NOTICE Inspect all connections and tubing before use and replace any defective parts Also inspect the column tube thoroughly before use to verify that there are no cracks or other visible signs of damage NOTICE Perform a leakage test whenever any of the wetted parts have been re fitted or changed Oo O OO gt gt NOTICE Always wear latex gloves or gloves of other suitable material when handling the bed support to prevent grease from unprotected fin gers coming into contact with the bed support umns User Manual 28 9562 89 AD 19 1 Introduction 1 6 Safety precautions NOTICE To avoid spillage do not open the bottom valve while the column is filled with liquid unless there are hoses connected to the outlets NOTICE Handle stainless steel parts especially the bed supports with care Damage to surfaces may lead to corrosion 1 7 Emergency stop procedures EMERGENCY STOP buttons Axi
111. mn and AxiChrom Master weights and dimensions on page 193 Inlet and outlet system e The tubing connection at the Top mobile phase on the adapter is the only liquid connection at the top of the column e Atthe bottom the center inlet outlet is the bottom mobile phase e The Slurry inlet is connected to the slurry tank from where the media slurry is drawn into the column Inside the Media valve under the column the Slurry inlet has a liquid connection with the Rinse inlet e The Rinse inlet is used to flush the Media valve the Slurry inlet and the tube to the slurry tank free from residual media after filling and unpacking the column When a filling process has ended the Media valve closes and a pump connected to the Rinse inlet pumps the liquid through the Media valve and then through the Slurry inlet to the slurry tank e The Slurry inlet and Rinse inlet have no connection to the Bottom mobile phase flowpath When the Media valve is open the Slurry inlet and Rinse inlet lead directly to the process chamber Liquid in the mobile phases on the other hand has to flow through the bed supports to reach the process chamber Connection flanges e Tri clamp 25 is used for the 300 400 450 and 600 columns e Tri clamp 50 is used for the 800 and 1000 columns Valve holder accessory The valve holder accessory is useful for fixing components to the column assembly particularly when a number of serially connected valves and sensors ar
112. mn is moved From cold to warm sur roundings The adapter dome nuts the tie rod bolts and the bottom bed support nuts should first be loosened to finger tight in the colder room and then moved to the warmer room When the column is properly tempered after about 24 hours the adapter dome nuts the tie rod bolts and the bottom bed support nuts should be tightened according to the tightening torques below From warm to cold sur roundings Ensure that the column is properly tempered after about 24 hours Then tighten the adapter dome nuts the tie rod bolts and the bottom bed support nuts according to the tightening torques below AxiChrom 300 1000 columns User Manual 28 9562 89 AD Tightening torques Tie rod bolts Tightening torques for tie rod bolts Column Tightening torque 300 400 and 450 20 Nm 600 28 Nm 800 42 Nm 1000 66 Nm AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 9 Ambient temperature changes 87 3 Getting started 3 9 Ambient temperature changes Adapter dome nuts and bottom bed support nuts Location of dome nuts Location of bottom bed support nuts l C O oS oe Ad Z LE FIELD IU GE Aff Tightening torques for adapter dome nuts and bottom
113. n how to unpack the columns Read Me First column from the delivery package Product documentationin Configured for A CD containing spare parts lists binder and on a CD individual diagrams material certificates columns etc AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 1 Introduction 1 2 Scope of this User Manual Additional documentation on request e Additional printed versions of documentation available electronically spare part lists diagrams and material certificates etc e Packing recommendations applications and Sanitization in place studies and other supporting documentation Other reference documents Other documents that should be referred to before using the column e AKTAprocess User Manual e AKTAprocess Operating Instructions e UNICORN User Reference Manual for AKTAprocess NOTICE Ensure that the product file is kept updated and together with the product documentation The file should contain information about inspections performed maintenance exchanged spare parts and any other important information for safe operation 1 3 Important user information Read this before using AxiChrom columns All users must read the entire AxiChrom 300 1000 columns Operating Instructions before installing using or maintaining the instrument Always keep the Operating Instructions at hand when using AxiChrom 300 1000 columns Do not operate AxiChrom 300 1000 columns in any other w
114. n the valve on the top mobile phase hose to release the Pressure in the column R is important to open the valve slowly if pressurized The adapter will start at a high speed to prime Arcam the top screen expelling liquid through the top Pre mobile phase Media valve closed Press CONFIRM to continue 1 Close the Seal flush valves 2 Stop the pump 3 Slowly open the valve on the Top mobile phase tubing to release the column pressure It is important to open the valve slowly when the column is pressurized When the valve has been opened direct the Top mobile phase to waste 4 Check the boxes and press CONFIRM when all the conditions are fulfilled The adapter will then start moving at a high speed to prime the top bed support expelling liquid through the Top mobile phase AxiChrom 300 1000 columns User Manual 28 9562 89 AD 105 4 Preparations before packing the column 4 4 Priming the column 4 4 5 Using Master wizard Step 12 wait for the adapter to move down Priming progress Mo ing the adapter down 11 15 een 4 53 romaning Adapter commanded posdion 30 0 om a Adapter position 54 4 cm SES Nee Moving the adapter down Pressure 1 05 bar Adapter moving Media valve closed The adapter now moves down The remaining time is displayed in a time bar Press PAUSE to stop the movement temporarily or to terminate the priming wizard Step 13 prime the bottom mobile phase Priming progress Prime the bottam mobile phase 1
115. n to the center of gravity label on the column stand Correct side view Correct top view Incorrect maka Wheel kits WARNING Columns in swing out or any kind of maintenance mode must not be moved This may cause personal injury or damage to the column CAUTION To avoid accidental collisions use caution when moving columns equipped with wheels by hand Two or more people may be re quired to move the column safely AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 3 Wheel kits NOTICE To avoid damage to the wheels and possible tipping when the column or AxiChrom Master is moved ensure that the wheels do not run into objects for example thresholds or tubing NOTICE Columns equipped with casters must be levelled properly with the casters removed before the column is used Wheel kits on AxiChrom 300 600 columns The wheels for 300 600 columns are delivered in a kit containing rear wheels lever trolley bar and trolley Different column sizes use different wheel kits NOTICE While transporting a 300 600 column do not pull the handle up ward as the trolley knob may fall out of its position Lifting the handle will not work as a brake A
116. nciples of column function Packing the column To pack the column the adapter moves downwards pushing down the slurry and forcing the liquid out through the bottom bed support and further out through the Bottom mobile phase Media particles are retained by the bed support and a bed is formed This is called the consolidation stage The pressure is monitored in the closed Top mobile phase flow path The pressure gradually increases while the bed is being consolidated After the bed is fully consolidated the adapter will eventually meet with the newly formed bed When this occurs the pressure drops momentarily in the column When the consolidated bed is detected either by the decrease in pressure or manually the operator is given an option to e Ignore if the detection was false Result The consolidation continues If no detection occurs the adapter stops at the defined Low bed height limit e Confirm the detection Result If the final bed height is calculated to be within the selected bed height range with the defined Packing Factor the adapter will continue to compress the bed When the defined Packing Factor has been achieved the op erator ends the packing procedure If the selected bed height range cannot be met the packing will be aborted The compressed bed should be within the defined compression range within the set bed height range to assure optimal compression and performance of the packed bed Unpacking the media from the co
117. needed for fitting and removing the wheels and trolley bar is stored in the box on the trolley Assembling the wheels on AxiChrom 800 and 1000 columns Note Note To be able to use the jack when assembling disassembling the wheels the distance between the floor and the column stand must be between 95 135 mm for AxiChrom 800 and 1000 columns The distance between the floor and the column stand is 122 mm when the wheels are assembled AxiChrom 300 1000 columns User Manual 28 9562 89 AD 53 3 Getting started 3 3 Wheel kits ic l ip ii oy 95 135 mm oy QW ADE AN KR ONSEN SKN NAN 1 Mount the wheels 2 on the underside of the wheel kit beam Mount the trolley handle 2 Use the handle on the trolley bar to steer it in between the front feet of the col umn The top B of the trolley bar should hang on the column stand when posi tioned Part Function B Trolley bar top C Lower part D Slot E Screw 54 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 3 Getting started 3 3 Wheel kits Remove the rear wheels from the wheel kit frame Place the jack under the frame at position I and use the jack to lever the front of the column so that the wheel kit can be assembled and tightened with the 16 mm wrench underneath the trolley bar G Place the brac
118. nsure that any liquids used with the column is particle free down to 1 um as particles can block and damage the bed support AxiChrom 300 1000 columns User Manual 28 9562 89 AD 17 1 Introduction 1 6 Safety precautions NOTICE Chlorides and low pH can cause corrosion on stainless steel Rinse thoroughly with clean water after use Inspect the column regularly for signs of corrosive attacks which may cause column damage if untreated Note that the stainless steel bed supports are especially vulnerable to corrosion NOTICE If there is air in the column ensure that there is an unrestricted flow path for evacuation through an open valve before the adapter is moved or liquid is pumped into the column O NOTICE Ensure that the pressure does not exceed the operating pressure of the media packed in the column NOTICE The acrylic column tube has limited resistance to organic solvents Refer to the Chemical resistance section for more information Maintenance WARNING Follow all safety instructions displayed in the AxiChrom Master when performing maintenance on the column 18 AxiChrom 300 1000 columns User Manual 28 9562 89 AD AxiChrom 300 1000 co 1 Introduction 1 6 Safety precautions WARNING Ensure that no body parts are caught between the column tube assembly and the column frame when using the swing out function for maintenance work on the colu
119. o classified according to USP Class VI 21 CFR Part 177 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 11 1 Introduction 1 4 Regulatory information CE Marking The CE marking and the corresponding Declaration of Conformity is valid for the instru ment when it is e used as a stand alone unit or e connected to other CE marked products recommended or described in the user documentation and e used inthe same state as it was delivered from GE Healthcare except for alterations described in the user documentation The Declaration of conformity is valid only for systems that are marked with the CE marking CE Regulatory compliance of connected equipment Any equipment connected to the AxiChrom 300 1000 columns and AxiChrom Master should meet the safety requirements of IEC EN 61010 1 IEC 61010 1 or relevant harmo nized standards Within EU connected equipment must be CE marked 1 5 Intended use of the columns AxiChrom columns are designed for process scale chromatographic separation of biomolecules such as proteins peptides and oligonucleutides AxiChrom columns can be operated at pressures up to 4 bar The columns must be used in accordance with the instructions and technical specifica tions in this manual and AxiChrom 300 1000 columns Operating Instructions to minimize the risk for injuries and damage to the columns or other equipment 12 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1
120. on The Master uses several different pressure limits during operation When a limit is reached the adapter will stop if moving and an alarm text will be displayed The limits are sum marized in the table below Sequence Limit Manual Low pressure 0 5 bar High pressure 4 bar Priming Low pressure 0 5 bar High pressure 4 bar Filling Low pressure 0 30 bar The adapter stops automatically at the limit and continues again when pressure rises above 0 25 bar High pressure 4 bar Packing Low pressure 0 5 bar Overpacking Overpacking safety stop 15 seconds after PF 0 03 is reached Bed detection 10 mbar pressure drop for more than 2 seconds editable Unpacking Low pressure 0 5 bar Bed expansion 0 30 bar The adapter stops automatically at the limit and continues again when pressure rises above 0 bar High pressure Regular unpacking for Capto Fast Flow and MabSelect media Bead size gt 50 um 0 5 bar The adapter stops automatically at the limit and continues again if the pressure falls below 0 2 bar High pressure II Advanced unpacking for High Performance me dia Bead size lt 50 um 1 0 bar The adapter stops automatically at the limit and continues again if the pressure falls below 0 5 bar 34 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 2 Specifications and characteristics 2 3 Primary specifications 2 3 4 Pressure and time out limits Sequence
121. on 5 Allow to stand at least overnight 6 Read the total volume and the media volume and calculate the slurry concen tration media volume total volume e Carefully tilt the cylinder and check the media surface If the surface does not sway the media is settled well enough If the surface is not sufficiently settled it is recommended to allow the sample to stand longer This occurs for some media especially small particles such as Sepharose High Performance but also for media that is settled in salt containing solutions 44 Priming the column In this section This section first gives an overview of the column priming procedure Later in this section there is a more detailed stepwise description of how to create and use a UNICORN method or Master to prime the column The procedure is described for columns with acrylic column tubes with deviations for stainless steel column tubes where appropriate 4 4 1 Short description of the priming procedure During the priming procedure the adapter will go to priming position at the top of the column The column is then filled with priming liquid When the priming liquid comes out from the column top the Seal flush valves see page 98 are opened and the pressure in the column is increased with the connected membrane valve When the pressure has been increased the adapter will move downwards past the priming groove expelling air from inside the column through the rinse channel out th
122. operating cycle CAUTION Do not use chemicals with temperatures above the specified limits 36 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 2 Specifications and characteristics 2 5 Chemical resistance NOTICE Chlorides and low pH can cause corrosion on stainless steel Rinse thoroughly with clean water after use Inspect the column regularly for signs of corrosive attacks which may cause column damage if untreated Note that the stainless steel bed supports are especially vulnerable to corrosion Note The information in Table has been collected from several published sources not from individual tests on column components It should be used only as a guide The effect of chemicals will generally be more severe at higher temper atures Note also that the combined effects ofagents have not been taken into account in this table The applicable chemical resistance depends on the con figuration of the column and the associated materials of construction Chemical Concen Time cycle Comments Operating tration 1 restrictions temperature Acetic acid 25 3h Cleaning In Place 2 C to 30 C 64 19 7 CIP Acetone 2 th Efficiency test 2 C to 30 C 67 64 1 Ammonium 2m3 5h Adsorption 2 C to 30 C 7783 20 2 sulphate Benzyl alco 2 12 months Storage 2 C to 30 C 100 51 6 hol Ethanol 20 12 months Storage 2 C to 30 C 64 17 5 and max 0 5 bar Ethanol 70 4 3h CIP 2 C
123. or example if the asymmetry factor is high or the bed stability is insufficient the packed bed can be further compressed manually This is providing that the resulting lower bed height is allowed Procedure 1 Enter the MANUAL CONTROL wizard from the MAIN MENU 2 Determine how much further to compress the bed Note that over compressing the bed may cause damage to the particles As a general recommendation the bed should not be compressed more than 0 05 higher than the recommended default value in the Master 3 Enter a low adapter speed 30 cm h is suggested 138 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 5 Creating custom packing methods in the Master 5 5 2 Post packing modifications Open the Bottom mobile phase pathway The Top mobile phase shall be closed Enter the lower target bed height and press return in the numeric pad Press RUN and the adapter will move down to the set height at the set speed expelling excess liquid through the Bottom mobile phase valve When the adapter stops close the bottom mobile phase pathway Evaluate the column performance see Chapter 6 Evaluating column performance on page 141 Method 2 If the bed was over compressed it can often be expanded in a controlled way Results are normally good but cannot be guaranteed Procedure 1 Start a liquid flow upflow through the column at least two times higher than the adapter speed in step 2 that is gt 60 cm h
124. packing method is used for High Performance media The number of slurry push out cycles in the High Performance unpack ing method depends on the bed height to be unpacked Two media push out cycles are used for beds lower than 25 cm and three cycles for beds higher than 25 cm Note Always prime the Media valve before operation AxiChrom 300 1000 columns User Manual 28 9562 89 AD 151 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes WARNING Ensure that nobody is present within the safety zone when the adapter is moving The operator in charge is responsible for keeping the safety zone free from unauthorized personnel and unnecessary equipment Step 1 Enter the Master unpacking wizard Welcome to the AxdChrom Master unpacking wizard This wizard will take you through a number of steps to confirm and edit the settings for the unpacking of your AxiChrom column Preconditions for unpacking are All hoses leading to the column are pnmed A pump or a hose from the system is connected to the Rinse Valve Both mobile phases are open A slurry tank is connected to the Slurry Inlet 1 Select UNPACKING in the MAIN MENU to enter the AxiChrom Master UNPACK ING wizard 2 Ensure that all preconditions are fulfilled before pressing CONTINUE All tubing leading to the column is primed A pump or a tube from the system is connected to the Rinse inlet Both mobile phases are opened A
125. primed see page 93 e The slurry concentration has been determined see page 91 Short description of the packing procedure The packing procedure starts with filling of the column with slurry from the slurry tank When the correct amount has been filled the adapter starts to go down consolidating the bed on the bottom bed support When all media have consolidated and the adapter meets the bed surface the bed contact is detected and the operator shall confirm the detection within 30 seconds After the detection the compression continues until the desired Packing Factor and or bed height has been reached The operator is then prompted to end the packing AxiChrom 300 1000 columns User Manual 28 9562 89 AD 109 5 Packing the column 5 2 Creating a UNICORN method 5 2 Creating a UNICORN method Background Procedure 110 The Method Wizard in UNICORN simplifies creation of an Intelligent Packing method The verified methods require an initial selection of a listed medium Thereafter the user makes appropriate selections to complete the creation of the method This section describes the creation of a UNICORN Intelligent Packing method If a custom medium is selected in the Method Wizard media list wider range of variables are allowed to be defined to optimize the method see Creating a packing method based on custom media on page 115 For information concerning low bed heights see Section 4 2 Preparation of the slurry on page 9
126. pture discs and safety equipment WARNING Do not block access to the power switch The power switch must always be easy to access CAUTION Do not touch the motor on the column during operation since the motor might be hot and there is a risk of burning injury CAUTION Do not stand on the column base to reach the top parts of the column e g top inlet outlet Always use proper equipment for climbing and standing when reaching the top parts of the column 16 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1 Introduction 1 6 Safety precautions CAUTION Remove any spillage on the floor immediately to minimize the risk of slipping accidents CAUTION Use a pressure gauge pressure relief valve rupture disc or other pressure safety equipment to ensure that the maximum operating pressure of the column is not exceeded NOTICE Do not operate the column at temperatures outside the operating temperature range NOTICE Ensure that the column and system is primed that is free from air and that the inlet is open before the process run is started NOTICE Do not use chemicals not listed in the Chemical Resistance infor mation O O O gt e The wetted parts of AxiChrom may be damaged by chemicals not listed in the Chemical Resistance information Contact your GE Healthcare representative before using chemicals that are not listed NOTICE gt E
127. r settings for the media are known Packing Factor packing speed and filling speed use the wizard for Intelligent Packing Procedure 1 In the Select media type dialog if the separation technique is known choose the technique otherwise choose All types 2 In the Select media dialog choose Other 3 In dialog no 6 15 press Edit to reach the template parameter settings They are deliberately set rather low for most media Press Set and enter the param eter settings found for the media 4 Proceed as usual according to the instructions in Section 5 4 Packing using the Master Wizard on page 120 Manual packing 136 Note This method is recommended for advanced users only swam ET Adapter Adapter position 10 3cm Direction Up Adapter speed 0 cmh Direction Down Gotoposition 40 0 cm f Adapter Speed 600 cmh Media valve closed Pressure 0 07 bar Adapter stopped MAIN MENU If the standard method does not fulfill the requirements the MANUAL CONTROL wizard can be used for packing either as it is or with the help of a standalone pump All interac tions need to be performed manually and without the help of the software interface Through manual control the following can be performed e Open or close the Media valve press OPEN or CLOSE e Control adapter movement press Jog Up or Jog Down AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 5 Creating custom packing method
128. r than the adapter speed The adapter speed can be increased in increments of 30 cm h up to 240 cm h e Ifthe vacuum for some reason is below 0 4 bar the adapter will stop The flow can be increased in steps of 25 cm h until the adapter starts to move again It is then possible to increase the flow to over 75 cm h until the pressure has reached the target pressure Use the arrow buttons to control the adapter speed e Press PAUSE to temporarily stop the action or to terminate the UNPACKING wizard and return to the MAIN MENU Step 9 Reverse the pump flow when the bed is expanded npacking progress Reverse flow 9 16 ane Capto Q 218 romaning ae Adapter position 14 0 cm Reverse the flow The bed has been expanded Reverse the flow at 377 nto move the expanded bed towards the bottom of the column Push the bed down fora total of 240 Pressure 0 03 bar Adapter stopped Press CONFIRM to continue Media valve closed CONFIRM The bed is now expanded AxiChrom 300 1000 columns User Manual 28 9562 89 AD 157 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes Reverse the flow through the column and run it at the flow stated in the dialog This will move the expanded bed towards the bottom of the column It is possible to continue the unpacking before the time has elapsed if the bed already has collapsed to the bottom of the column Run the pump for the time stated in the dialog Chec
129. rom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 7 Using the Master 3 7 3 Column and Master setup e Press CONTINUE when a column is chosen in the list e Press CONFIGURATION to enter the Configuration wizard from where it is possible to view system settings system specifications and test the system If the settings are corrupted If the HELP button turns blue when a column is chosen in the list the settings for the column have been corrupted It is not possible to continue using the settings for this column when they are corrupted This situation can occur when the AxiChrom Master system was last shut down and was not allowed to save the data completely before the Power switch was turned off If this happens follow the instructions below to recreate the column settings Press the HELP button for more information Recreating column settings 1 Press Delete to remove the corrupted column settings 2 Press Add and create new column settings 3 Choose the column parameters for the deleted column in the list Type a name and press CONTINUE 4 Verify the column parameters according to the wizard instructions 5 Calibrate the motor according to the instructions in the Master 6 Verify the adapter position according to the wizard instructions 1 The button also turns blue when the selected line is empty AxiChrom 300 1000 columns User Manual 28 9562 89 AD 79 3 Getting started 3 7 Using the Master 3 7 3 Co
130. rough the Seal flush valves AxiChrom 300 1000 columns User Manual 28 9562 89 AD 93 4 Preparations before packing the column 4 4 Priming the column 4 4 1 Short description of the priming procedure When the adapter stops the Seal flush valves are closed and the membrane valve fully opened The adapter then starts to move and the top bed support is primed When the adapter passes 30 cm from the bottom bed support the priming direction changes and the bottom bed support is primed When the adapter comes to the bottom of the column and stops the priming procedure is finished Priming is performed either using UNICORN on an AKTAprocess system or from the Axi Chrom Master directly The workflows are described in Section 4 4 4 Using UNICORN method on page 97 and Section 4 4 5 Using Master wizard on page 99 4 4 2 Wetting plastic bed supports Follow the instructions below if the column is fitted with plastic bed supports Note Before priming the column mount the bed supports dry and assemble the column as described in the AxiChrom 300 1000 Operating Instructions Connect membrane valves on the column top and bottom mobile phases Step Action 1 Prime the column according to the AxiChrom Master priming instructions or a UNICORN priming method 2 Perform a leakage test 3 With the adapter at 1 cm from the bottom bed support equilibrate the col umn with 20 v v 1 propanol at 30 cm h under a pressure of 1 bar for five column volu
131. rther explanations of the progress dialogs in the wizards see Information in progress dialogs on page 78 Step 1 prime the media valve manually If the column is primed with the guidance from the Master and a standalone pump the Media valve has to be primed manually Follow the procedure below to prime the Media valve manually AxiChrom 300 1000 columns User Manual 28 9562 89 AD 99 4 Preparations before packing the column 4 4 Priming the column 4 4 5 Using Master wizard 100 1 Connect a pump to the Rinse inlet 2 Connect the slurry tubing tank to the Slurry inlet 3 Open the slurry tubing tank 4 Close the Bottom mobile phase and the Media valve 5 Start a flow through the Rinse inlet and further into the slurry tubing tank at approximately e 200 I h for 300 400 450 and 600 columns e 800 I h for 800 and 1000 columns 6 When the tubing is free of air make sure the Top mobile phase is open and then open and close the Media valve from MANUAL CONTROL in the Master a couple of times To make sure no more air comes from the Media valve continue to pump liquid through the valve filling the bottom of the column and covering the bottom bed support with liquid for example 1 cm If the column is packed avoid opening the Media valve during the valve priming 7 When no more air comes up from the Media valve close it 8 Stop the pump and close the slurry tank 9 The Media valve has now been primed 10 If applicable calcula
132. ry concentration before the column is repacked see Section 4 3 Determining the slurry concentration on page 91 2 Press MAIN MENU to leave the UNPACKING wizard 7 3 4 Unpacking stainless steel column tubes Unpacking stainless steel column tubes using a UNICORN method Columns in stainless steel column tubes are unpacked using a UNICORN method in the same way as for acrylic column tubes see Section 7 3 in the AxiChrom 300 1000 User Manual edition AB Unpacking stainless steel column tubes using the Master wizard For the general unpacking procedure using a UNICORN method see Section 7 3 in the AxiChrom 300 1000 User Manual edition AB With stainless steel column tubes it is not possible to see when the column bed expands step 8 9 The unpacking progress can however be followed from the pressure on sensor PRESS_119 The pressure drops when adapter starts to move upwards and the bed ex pands under upflow then rises again abruptly when the adapter is in the top position and the flow is reversed There will be a further slower increase in pressure as media settles on the bottom bed support Go to the next breakpoint in the unpacking method when the pressure increase indicates that medium is accumulating on the bed support 162 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 3 Unpacking the column 7 3 4 Unpacking stainless steel column tubes page 7 1 1 The pressure blue trace drops as the
133. s With Plastic bed SUPPOFES wacecceccccsssssssseesessssscccssssssssusssssessesssssssssusssessesseessssssssassseesees 177 A 4 Columns with stainless steel bed SUPPOFTS e seeeesssesssssessssssneessssenssssnsssssstsssestsseeseesnceeeeneeeeees 179 A5 Media Valvecind tubes iiccaiuecclidnduhaldentiansenaan dees 181 Packing Duffer MUU IE RCN A accep htc costa teeters eee 183 Accessories and additional literature ssssssesssssessssssesssssssesssssressssesseseessssse 187 Crate weights and dimensions s sssseessssssessssessesssessessesesseseesesssessessssessesssssses 192 Column and AxiChrom Master weights and dimensions cscsceceeeeees 193 DECONTAMINATION repor tssirini eera enin asiri eri ei 195 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 1 Introduction 1 Introduction 1 1 General introduction AxiChrom process column family has been designed to deliver reproducible results from process development to production scales This is facilitated by the innovative Intelligent Packing where UNICORN software AKTA systems and AxiChrom columns work to gether to facilitate a convenient operation for packing of the bed via axial compression The AxiChrom family of process column comprises the following columns e AxiChrom 300 e AxiChrom 400 e AxiChrom 450 e AxiChrom 600 e AxiChrom 800 e AxiChrom 1000 The collective name AxiChrom 300 1000 refers to all columns in the family Columns are available in t
134. s are Requirement Value Supply voltage 380 400 VAC Nominal current 10 15 16 A NTD Non Time Delay min max Frequency 50 60 Hz Max voltage North America 480 Y 277 VAC Max current 6A Max power consumption 2400 VA Short circuit rating 5 kA Note Detailed requirements may vary for different column configurations Refer to the column documentation for the applicable requirements for your specific column and AxiChrom Master The power cables for both the AxiChrom Master and for the adapter control motor are delivered attached to the AxiChrom Master The system shall be connected to a 400 V supply voltage system or a 480 V supply voltage system with a maximum voltage of 277 V between phase and ground A suitable con nector has to be selected and assembled on the AxiChrom Master by the customer to comply with the supply voltage system and local regulations The Motor Power cable for the adapter control motor is permanently installed on the AxiChrom Master The motor feedback communication cable is connected to Motor Feedback Both these cables must be connected to the control motor before AxiChrom Master is connected to the power outlet Color coding of cable conductors Conductor Color Protective ground earth Green yellow Live 1 No 1 or Black Live 2 No 2 or Brown Live 3 No 3 or Grey The black brown and grey conductors may be connected to any of Live 1 2 and 3 The
135. s are met and clearly understood Notices NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment AxiChrom 300 1000 columns User Manual 28 9562 89 AD 9 1 Introduction 1 3 Important user information Notes and tips Note A Note is used to indicate information that is important for trouble free and optimal use of the product Tip A tip contains useful information that can improve or optimize your procedures Typographical conventions Software items are identified in the text by bold italic text A colon separates menu levels thus File Open refers to the Open command in the File menu Hardware items are identified in the text by bold text e g Power switch 1 4 Regulatory information This section describes the directives and standards that are fulfilled by the AxiChrom 300 1000 columns and AxiChrom Master Manufacturing information Name of manufacturer Address of manufacturer GE Healthcare Bio Sciences AB GE Healthcare Bio Sciences AB Bj rkgatan 30 SE 751 84 Uppsala Sweden Other regulatory information may be found in the Declaration of Conformity supplied with the product CE Conformity This product complies with the European directives listed in the table by fulfilling the corresponding harmonized standards A copy of the Declaration of Conformity is supplied with the product Directive Title 2006 42 E
136. s in the Master 5 5 1 Packing stainless steel column tubes e Set an adapter position press Set and then RUN to start the adapter movement Press STOP to stop the movement The position can be changed again while the adapter moves If the position is changed during adapter movement the RUN button will be lit up To verify that the position is to be changed press the RUN button e Change the speed press Set The adapter speed can be varied from 10 to 600 cm h There are two height limits in the manual mode for the adapter The lower limit is at 1 cm from the bottom The upper limit is the priming position above the priming groove which should be used for priming the column Maximum filling heights are listed in the table below If the adapter is forced past a limit the adapter will stop automatically When filling the column manually pay attention not to move the adapter past the priming groove 300 600 columns 800 1000 columns Short column tubes 570mm 578 mm bed height 100 300 mm Long column tubes 830 mm 838 mm bed height 100 500 mm 5 5 1 Packing stainless steel column tubes Packing the column using a UNICORN method For the general packing procedure using a UNICORN method see Section 5 3 Packing using a UNICORN method on page 117 With stainless steel column tubes it is not possible to see when the adapter is approach ing the consolidated bed surface step 7 In order to verify bed detection observe t
137. se description of how to use a UNICORN method or the Master to unpack the media from the column Recommended unpacking solution The recommended unpacking solution is water If the media is to be stored unpack in appropriate storage solution such as 20 ethanol 0 2 M sodium acetate in 20 ethanol or 10 mM NaOH AxiChrom 300 1000 columns User Manual 28 9562 89 AD 145 7 After the packing 7 3 Unpacking the column 7 3 1 Short description of the unpacking procedure 7 3 1 Short description of the unpacking procedure There are two unpacking procedures that can be used for unpacking media from the column If unpacking a Bioprocess media listed in UNICORN or the Master the correct method is chosen automatically The different procedures are the regular procedure described below used for e g MabSelect Capto and Fast Flow media and the advanced method for e g gel filtration and High Performance media The difference between the methods is flow direction in the expansion step and number of repetitions of media push out steps If custom media is to be unpacked see Creating a custom unpacking method on page 149 The regular unpacking procedure consists of three steps expansion collapse and slurry push out The expansion is performed by lifting the bed by moving the adapter up at the same time as flow comes from the bottom bed support When the bed has expanded the flow is reversed to collapse the bed by pushing it down When the collapsed bed
138. sing START The preconditions are the same as when selecting PRIMING from the MAIN MENU AxiChrom 300 1000 columns User Manual 28 9562 89 AD 101 4 Preparations before packing the column 4 4 Priming the column 4 4 5 Using Master wizard Step 5 wait for the adapter to move up Priming progress Moving the adapter up 4 15 5 35 romaning Moving the adapter up Adapter is moving to priming position Pressure 0 04 bar Adapter moving Media valve closed i iili ilij gi The adapter will now move upward to priming position above the priming groove If the movement needs to be stopped temporarily press PAUSE Priming progress Paused een Priming paused Adapter position 10 3 cm The Priming has been paused To abort the Priming procedure and continue manually press ABORT Press CONTINUE to resume the Priming process Pressure 0 07 bar Adapter copped Media valve closed ABORT The priming wizard can also be terminated from here 102 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 4 Priming the column 4 4 5 Using Master wizard Step 6 start the pump upflow Priming progress Start the pump upfiow 5 15 ena 15 35 remaining ae Adapter position 61 8 cm Start the pump upflow Start an upwards flow through the J Column at 1800 Vh Run the pump for a total of 16 15 until liquid comes out through the top mobile phase hose Pressure 0 32 bar Pre
139. slurry tank is connected to the Slurry inlet 152 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes Step 2 Select media type Select media type 2 16 Giren Media type All types ffi A ranne Cation exchange Metal affinity Hydrophobic interaction Size exclusion pamm Et cosa ne BECP CONTINUE aL Select a media type for the chromatography technique that is packed in the column using the arrow buttons The selection All types will list all available media regardless of their applications 2 Press the CONTINUE button to proceed Step 3 Select media Select medium 3 16 awam Medium Capto Adhere Capto DEAE Q Sepharose Big Beads Q Sepharose Fast Flow Q Sepharose High Performance Q Sepharose XL Other 1 Select the packed media for the selected chromatography technique that was picked in the previous dialog A scroll bar will appear on the right side of the list if it is too long to fit on the screen All lists include a selection called Other which is to be used for media other than Bioprocess media from GE Healthcare 2 Press the CONTINUE button to proceed AxiChrom 300 1000 columns User Manual 28 9562 89 AD 153 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes Step 4 Verify bed height Pte ti Capto Q Bed height 10 0 cm 1 Check that the bed height is corre
140. ss CONFIRM to continue Adapter stopped Media valve closed 1 Start an upward flow i e from the bottom to the top through the column at the stated flow displayed in the dialog The time on the time bar will start when the first box has been checked 2 Run the pump for the stated time displayed in the dialog or until liquid comes out through the tubing on the Top mobile phase 3 Check the boxes and press CONFIRM to continue Step 7 wait for the buffer to pass the priming groove Priming progress Buffer have passed the priming groove 6 15 The liquid level is above the priming groo Adapter position 61 8 cm q Aj ag ng z The kquid level is above the pnming J groove Press CONFIRM to continue Pressure 0 04 bar Adapter stopped Media valve closed CONFIRM 1 Check the dialog box when the liquid is above the priming groove found on the inside in the top of the column tube 2 Press CONFIRM to continue AxiChrom 300 1000 columns User Manual 28 9562 89 AD 103 4 Preparations before packing the column 4 4 Priming the column 4 4 5 Using Master wizard 104 Step 8 reduce the flow Priming progress Reduce flow 7 15 awam Reduce flow Adapter position 61 8 cm Set flow to 402 th m Press CONFIRM to continue Pressure 0 31 bar Adapter stopped Media valve closed Set the flow to the value stated in the dialog 100 cm h Check the box when completed Press CONFIRM to proceed
141. ssesssecceseen i 33 2 3 4 Pressure and time out limits eeeeesessssssssssssssssssssssssssssssssssssssssssssssssissnntntttinitsntntsnnenseeneneeee 34 24A MAENG cenaa ea e e RE 35 2 5 Chemical resistance u 36 2 5 1 Resistance to sodium chloride 39 3 Getting Se ccc asc ca aa eg i odie secretin ai ce eccceeseta dente eaten eens 40 3 1 Unpacking the crates and cleaning a NEW COLUMN csssssssssessessccsssssssssssseesessccsssssssessssesees 40 3 2 Placing the column ANd Master ceessscssssssssssssssseeeseesee w 42 3 3 Wheel kits Ss 46 3 4 Connections a 59 35 RUPES Smesna E E a PAE 66 36 Colummandisystem Set Pssnssspsnainsssnsuansnean aaa 68 3 7 Using the Master o ecccssssssessscssccssccssssesssees 75 3 7 1 Start Master and calibrate column 75 3 7 2 User interface sssri m 76 373 Column and Master Set p oermeniniiniiinnnnaa 78 38 Cleaning th COMMA eisino inanan n AA AAAA T 85 39 AmMbienttemperature CHOMGE S sesssscesatsassiivisvsensioacssvetessaassbvatelacassdaveupciasstscsnvvvnsnonteesssbbtelecCasastonsl 86 4 Preparations before packing the column seessssssessssssressssessesssessesessesseseeesssse 89 4 1 Packing Factor and Compression FACtOT cssssssssssssssscssssssssssssesesssseesessssssssusssesssccessssssssunseesees 89 4 2 Preparation of the SIUFY sssssssssssssssssssisssrssrsssssstssssesssssnenstssrnnsastssntrntnnnnentnsnntantnnntnnnarstrnnrnnnnnnnnnnrns 90 4 3 Determining the slurry con
142. t bottom mobile phase gt top mobile phase gt waste upflow valve 6 pump gt bottom mobile phase gt top mobile phase gt closed gt Unpacking me media valve open to slurry tank dia push out and down flow pump gt valve 2 gt top mobile phase gt bottom mobile phase gt waste valve 6 70 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 6 Column and system setup Manual configuration minimal setup The following manual setup makes it possible to perform the PRIMING and INTELLIGENT PACKING procedures available in the AxiChrom Master The setup comprises two way valves and shut off valves to utilize different flow paths for the different procedures For symbol explanations see the table on page 69 Slurry Buffer Slurry in To waste To waste Possible flowpaths for different column operations Process Flow path alternatives from gt to Pump gt bottom mobile phase gt top mobile phase gt waste valve 5 Priming Column gt bottom mobile phase gt waste valve 6 Packing Column gt bottom mobile phase gt waste valve 6 Rinse Pump gt valve 2 gt rinse gt slurry tank AxiChrom 300 1000 columns User Manual 28 9562 89 AD 71 3 Getting started 3 6 Column and system setup Automatic configuration connecting AKTAprocess The following setup makes it possible to perform all procedures that are available in the Me
143. t e 5 5 to 7 bar The pneumatic air supply connections on AxiChrom Master are illustrated in Connections on Master and column on page 59 3 5 Rupture discs 66 To secure equipment and personal safety rupture discs are available as accessories Note Installation of rupture discs will change the maximum operating pressure for the column from 4 to 3 8 bar g due to rupture disc characteristics For column sizes 300 600 there is a special T junction provided for use with rupture discs Use this junction only with rupture discs recommended by GE Healthcare For column sizes 800 1000 use a standard 1 T junction part no 28938170 Description 1 T junction 3 Vent 2 Rupture disc 4 Clamp Figure 3 2 Rupture disc assembly AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 5 Rupture discs Installing rupture discs NOTICE Rupture discs should be installed at both mobile phases To install rupture discs do as follows 1 Connect the T junctions to the column mobile phases according to Section 3 6 Column and system setup on page 68 2 Choose appropriate hose or pipe with a TC50 ferrule as vent CAUTION Do not place rupture disc vent towards an area where it would endanger personnel 3 Verify and ensure that all ferrule and clamp working surfaces are clean dry and free of nicks dents gouges and wear 4 Inspect gasket position and ensure that gasket ID is c
144. t of media in the Method Wizard for Intelligent Packing and in the Axi Chrom Master For media not included in the Method Wizard it is possible to create a new packing method in UNICORN see page 110 or in the AxiChrom Master see page 135 Intelligent Packing procedure Intelligent Packing uses interactive guides to ensure an optimized compression of the bed Intelligent Packing will pack the column with minimal manual intervention to repro ducible results When the bed has been detected the adapter will proceed to compress the bed until the user is guided to stop the adapter at a specific Packing Factor which is determined to provide the best results for the selected BioProcess media 108 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 1 Intelligent Packing NOTICE In the Intelligent Packing tests performed by GE Healthcare on its base media selection the default Packing Factor for each media has been used and verified The user is therefore recommended to select the default Packing Factor For information change the default packing factor see Section 4 1 Packing Factor and Compression Factor on page 89 Prerequisites for Intelligent Packing e The column is connected to an Axichrom Master unit e The column is properly connected to an KTAprocess system or has a manual con figuration see page 68 e The start up instructions on page 75 have been followed e The column is
145. t upflow and Packing test downflow Select whether to perform packing tests up flow downflow or both Both are selected by default Inlet for sample Select the appropriate inlet for sample solu tion 114 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 2 Creating a UNICORN method 6 Click Next to display the Column Packing Summary Method Wizard page Mulliud AE fur arale Jutta Dukan Aiur bUn UUI Make sure that the system COLUMN TOP Connected 10 the cutlet on the side of the OF velve Corraci the AudChicm Macie preemie tarama PIS_119 to ayitem INLET CIPI Connect the cohn mobile phase top to the Aad hecen Mosier crersuse tarumtier PIS 119 This page provides a summary of the options selected in the previous wizard pages containing for example e estimated slurry volume based on the entered estimated concentration e estimated consumption of buffer and sample e instructions on how to connect the AxiChrom column to the AKTAprocess system 7 Click Finish The column packing summary can then be viewed under View Run Setup Notes Method Notes in the Method Editor module of UNICORN 8 Choose an appropriate result file folder result name and save the method Creating a packing method based on custom media Intelligent Packing methods may also be developed for custom media using the Method Wizard The wizard facilitates this by providing an extended range of variab
146. te the new slurry concentration by adding the amount of packing buffer pumped into the slurry tank to the total volume in the tank Note To minimize the dilution of the slurry in the slurry container prime the Media valve and slurry tubing with packing buffer before the slurry container is con nected Then prime the connection with a minimal amount of packing buffer but enough to get rid of any air in the connection Step 2 select PRIMING from the MAIN MENU The column is primed when there is no air inside the column and connected tubing To achieve this follow the instructions in the PRIMING wizard in the AxiChrom Master Press CONTINUE to start the priming procedure Note Ifthe column has not been used for some time add or change the 20 ethanol between the scraper seals Preconditions for priming are e Apump or a system e g AKTAprocess system is connected to the Bottom mobile phase e Both the Top mobile phase and Bottom mobile phase are open e The Seal flush valves are open and directed to waste AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 4 Priming the column 4 4 5 Using Master wizard e Amembrane valve is connected on the outlet of the system used during priming or on the Top mobile phase between the system and the AxiChrom Master pressure transmitter PIS_119 The membrane valve shall be open e The tubing on the Top mobile phase is directed to waste Not
147. tely flushed out and replaced by an appropriate storage solution after priming the column Result The column is now primed and cleaned AxiChrom 300 1000 columns User Manual 28 9562 89 AD 85 3 Getting started 3 8 Cleaning the column Cleaning the Master Turn off the AxiChrom Master before it is cleaned with water and or 20 v v ethanol 59 Effects of temperature changes Ambient temperature changes If the temperature surrounding the column is changed significantly the acrylic tube will shrink or expand This will cause unwanted tension in the acrylic tube in the dome nuts on the adapter the bolts of the tie rods and the bottom bed support nuts i e they will become tightened or loosened The tightening torque of the adapter dome nuts tie rod bolts and bottom bed support nuts must be checked whenever the temperature has changed see Tightening torques on page 87 The bolts should be tightened crosswise in small intervals Make sure that the torque wrench is calibrated and that the correct socket is used General guidelines 86 e Thecolumn should not be placed in direct sunlight since this could affect the column temperature e The column should preferably be packed and run in a room without significant temperature changes e Check the tightening torques of the adapter dome nuts tie rod bolts and bottom bed support nuts when the column is new and has been placed in its first location If the colu
148. ternatively 0 8 M sodium chloride or equivalent can be applied to a 0 4 M NaCl elution buffer depending on the media that is used Use a fluid velocity 30 to 40 cm h for 34 um and 50 um media and 20 cm h for 75 and 90 um media Running the HETP test at a higher velocity or using a larger sample volume will affect both HETP and AS values The airtrap and any filters should be bypassed during the sample application to avoid back mixing An application note describing column testing is available from GE Healthcare HETP calculation 142 Calculate the height equivalent of a theoretical plate HETP and asymmetry factor As from the chromatogram as follows HETP L N N 5 54 V W Variable Meaning L Bed height N Plate number Ve Retention volume Wh Peak width at half peak height Vp and w are in the same units To facilitate comparison of column performance the concept of reduced plate height h is often used Reduced plate height is calculated as h HETP dp where dp is the mean diameter of the beads As a guideline a value of h lt 3 is normally well acceptable at the optimal test conditions presented above AxiChrom 300 1000 columns User Manual 28 9562 89 AD 6 Evaluating column performance 6 2 HETP and asymmetry calculation Asymmetry calculation The peak should be symmetrical and the asymmetry factor as close to 1 as possible values between 0 8 and 1 8 are usually acceptable A change
149. th Press CONFIRM to finalize the priming procedure Adapter position 1 0 om Pressure 0 04 bar Adapter stopped Media valve closed 1 Close the Bottom mobile phase and then check the box in the dialog 2 Press CONFIRM to finalize the priming procedure Step 16 finalize the priming i The column is now primed Press MAIN MENU to end the priming wizard 2 Exchange the liquid in between the scraper seals to 20 ethanol AxiChrom 300 1000 columns User Manual 28 9562 89 AD 107 5 Packing the column 5 Packing the column In this chapter This section contains a brief overview of Intelligent Packing and stepwise descriptions of how to create and use a UNICORN method or AxiChrom Master wizard to pack media inthe column Columns are packed according to Intelligent Packing procedures regardless of whether a UNICORN method or Master wizard is used General instructions deal with packing acrylic column tubes procedures for stainless steel column tubes are described in Section 5 5 1 Packing stainless steel column tubes on page 137 5 1 Intelligent Packing Introduction Intelligent Packing allows for convenient and precise control of the column packing using an AxiChrom column together with an AKTA system UNICORN software and media with verified packing methods Several media are available from GE Healthcare that have verified packing methods for use with AxiChrom column and Intelligent Packing These are found in a lis
150. that the cable barrel is straight and engages correctly in the threads of the connector Remove the protective cap from the Profibus connector of the Master unit Connect the Profibus cable to the Profibus connector of the Master unit Power on and set up the system 1 Power on the AKTAprocess unit and allow 5 to 10 seconds for the boot process to finish Power on the Master unit Log in to UNICORN n system control in UNICORN disconnect AKTAprocess n administration system setup in UNICORN select the component AxiChrom 300 1000 for the AKTAprocess system n system control in UNICORN connect the AKTAprocess system On the screen of AxiChrom Master select the appropriate column to be used and confirm the bed height Exit to the MAIN MENU in the AxiChrom Master unit The system is now ready to use Disconnecting AKTAprocess from AxiChrom Master 1 2 74 In system control in UNICORN disconnect KTAprocess Power down the KTAprocess unit AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 6 Column and system setup 5 CAUTION Make sure that the AKTAprocess unit is powered off before the following step is performed Power down the AxiChrom Master unit with the Power Off button in the MAIN MENU 4 Disconnect the Profibus cable from the AKTAprocess unit 5 Place the Profibus termination plug on the connector of the AKTAprocess unit 6 Remove the Profibus cable fro
151. the appropriate variables siid Har fur syal MlA piuks atrliuus bUn Packing Parameters 1 Taget bed height 5 030 cm Recommended at least 10 0 cm Iriet for mobie phase Art irem chametes of shay hove 0 1 1000 mm 23 Length of shary hose 0 101000j m 2 AxiChrom Caa C _ Coa Ca Setting Description Target bed height Enter a value within the designated range Inlet for mobile phase Select the appropriate inlet that will be used for equilibration buffer when evaluating the efficiency of the packed bed in the column Inner diameter of slurry Select the appropriate diameter hose Length of slurry hose Select the appropriate length AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 2 Creating a UNICORN method 4 Click Next to display the Packing Parameters 2 Method Wizard page ASUS Wika fur sgela AEA bruise AGUI LONLO Packing Parameters 2 E Ure delat Packing Factor Packing tector 110 Min bed hang kmi 4 0 20 0 com Max bed night kest 20 0425 0 om 120 no AxiChrom ebat Nets Crea Hep Setting Description Min bed height limit Enter the appropriate height Inlet for mobile Enter the appropriate height phase Use default Packing This box is checked automatically which means Factor that the medium will be packed using a predeter mined Packing Factor Unless specifically stated in the instructions for a particular medium
152. the column Note The column is delivered partially filled with 20 isopropyl alcohol ethanol for conservation Make sure that this solution is washed out according to the in structions below WARNING Only personnel with appropriate training may move and unpack the delivery crates All movements and unpacking must be per formed in adherence with local regulations Even if the instructions in the Safety Instructions and User Manual are followed it is the customers responsibility to guarantee the safety of the personnel working with AxiChrom columns WARNING The column packing crate is marked with the center of gravity Ensure that the crate is properly balanced so that it will not acci dentally tip when moved 40 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 1 Unpacking the crates and cleaning a new column Cleaning a new column The column is delivered partially filled with 20 v v ethanol storage solution for con servation The column must be unpacked from the crate and washed as soon as possible after delivery preferably within 3 months Please make sure that the column is in correct condition and free from damage according to the instructions in Read Me First WARNING Always use protective glasses and other personal protective equipment appropriate to the current application to ensure per sonal safety during operation hic Then The column
153. the column to a pump that can deliver a flow of 100 cm h for the specific column used Wash the media up flow with pure water for 5 column volumes Change flow direction from up flow to down flow Let the media consolidate in 1 column volume of pure water at 100 cm h Determining the slurry concentration 16 17 Let the gel bed stabilize for 30 minutes without any flow and then read the bed height Convert the bed height to slurry concentration slurry concentration bed height slurry height 100 e g a bed height of 58 mm obtained from slurry filled to 100 mm gives a slurry concentration of 58 Remember to adjust for dilution step 8 if required e g if 20 mm water is added to the column first and slurry is added to a final level of 100 mm divide the calculated slurry concentration by 0 8 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 4 Preparations before packing the column 4 3 Determining the slurry concentration Gravity settling Gravity settling in a graduated cylinder is one of the most commonly used methods It is best to use fairly large cylinders for process scale columns at least 1 liter of slurry 1 Stir the sample so that all media is suspended and the slurry is completely ho mogenous 2 Quickly remove a sample using a container for example a beaker 3 Pour it immediately into the cylinder 4 Place it on a flat surface and cover the opening with for example Parafilm to minimize evaporati
154. the pump can be used for both upflow and downflow connected to either the Top or Bottom mobile phase valves see Section 3 6 Column and system setup on page 68 It must also provide the velocities in the expansion phase stated in the dialog listed in the table below Table 7 1 Conversion of flow velocities to volumetric flow rates for 30 and 75 cm h Column diameter mm Area cm L h 30cm h 75cm h 300 707 21 53 400 1257 38 94 450 1590 48 119 600 2827 85 212 800 5027 151 377 1000 7845 236 589 Step 6 Fulfill the conditions before unpacking Before the unpacking is started the following preconditions need to be fulfilled AxiChrom 300 1000 columns User Manual 28 9562 89 AD 155 7 After the packing 7 3 Unpacking the column 7 3 3 Unpacking acrylic column tubes 156 Preconditions 6 16 Sous Capto Q Before you can start unpacking the following preconditions need to be fulfilled The hoses and the system pump is primed J Hoses to the column are primed i The flow path to the column is open J A Slurry tank is Connected to the Slurry Inlet Approximate required buffer volume 101 L mis The required buffer volume is displayed in this dialog Make sure that this volume is available Ensure that e The external system pump is primed Ensure that the pump can be used for both up and downflow connected to either the Top or Bottom mobile phase valves e All tubing to t
155. the trolley to one of the sides as the following image shows Lower the front of the column by raising the handle on the trolley The handle should now stand vertically 3 If necessary remove the safety pin and loosen the height adjusting screw to be able to relieve the weight from the trolley knob D Remove the trolley 4 Unscrew the bolts B and remove the trolley bar from the front of the column stand 5 Place the lever close to one of the rear feet and wheels and lift the column by lowering the lever arm If needed in narrow spaces the lever can be mirrored by changing the attachment of the lever arm 52 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 10 3 Getting started 3 3 Wheel kits Remove the washer and safety pin from the wheels and disassemble the wheels from the hole A in the column stand 90 110 mm LL LLALL ELL L EL LLL 7 Lower the column by lifting the lever arm Remove the lever Repeat steps 4 to 6 on the other rear foot Ensure that the column stands firmly on all feet Ensure that the column is level see Placing the column on page 42 Wheel kits on AxiChrom 800 and 1000 columns The wheels for 800 and 1000 columns are delivered in a kit containing a jack trolley bar bracket and tools Different column sizes use different wheel kits All equipment
156. thod wizard in UNICORN Priming Intelligent Packing and Unpacking The setup is simplified by using AKTAprocess controlling the pump and valves for flow directions For symbol explanations see the table on page 69 Outlet 1 waste Outlet 2 blocked by valve blind flange Slurry Column 1 top valve X Column pottom valve PO O ce AKTAprocess system CIP 1 inlet CIP 2 inlet Bottom mobile phase Column 1 top valve Profibus connection Column 1 bottom valve CIP 1 inlet Side port of CIP Column Pack Valve block CIP 2 inlet 72 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 6 Column and system setup Connecting to an AKTAprocess system To simplify automated priming column packing with HETP evaluation or column unpack ing procedure the AxiChrom column has to be connected to a AKTAprocess system equipped with the optional CIP Column Packing valve block as described below For connections on AxiChrom Master and column see Section 3 4 Connections on page 59 Note e Use the Column valves If the AKTAprocess system is equipped with two column valve blocks e During priming a membrane valve should be attached to the top mobile phase Procedure il Connect a short piece of tubing of suitable length and the same inner diameter as the system from Column top valve to Side port of the CIP Column Pack valve block Connect the AxiChrom Master pressure trans
157. ting velocity of the process but preferably more 2 Continue to slowly increase the flow until a small head space is formed in the top or bottom of the column Make sure that the pressure rating of the column or the media is not surpassed 3 Maintain this velocity for 3 column volumes and then slowly lower the velocity again Make sure that there is no sudden decrease in the flow since this may damage the structure of the bed 4 If necessary reverse the flow and repeat the procedure AxiChrom 300 1000 columns User Manual 28 9562 89 AD 6 Evaluating column performance 6 Evaluating column performance Introduction The efficiency of a column depends on for example how well it is packed A poorly packed column gives rise to uneven flow resulting in zone broadening and reduced resolution It is therefore important to have a method by which the column can be tested before it is put into operation Such a method should be simple quantitative and should not introduce contaminating materials It is also an advantage if the same method can be used to monitor column performance over its working life so that it is easy to determine when the medium should be repacked or replaced Experience has shown that the best method of expressing the efficiency of a packed bed is in terms of the height equivalent to a theoretical plate HETP and the peak asymmetry factor A These values can be determined easily by applying a sample of for example ace
158. tion has taken place 8 If the detection of the consolidated bed surface is correct click Continue and be prepared to end the packing Result The bed compresses and the End Packing figure in the UNICORN flow scheme is colorized PF 1 06 Target PF 1 10 Time to PFs 47 Goto End Pack The compression can be followed as the colored boxes light up in brighter red or green color depending on the position of the adapter When the position of the adapter is within the approved bed height range and Packing Factor range the Goto End Pack button lights up green PF 1 09 Target PF 1 16 Time to PFs 12 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 119 5 Packing the column 5 3 Packing using a UNICORN method 9 When the desired Packing Factor is approaching double click the Goto End Pack button Result The manual control pump command window is opened with End Pack command highlighted Note Goto End Pack can be selected by double clicking at any time during the com pression step 10 To stop the adapter when the optimal Packing Factor or desired bed height is reached click on Execute to execute the End Pack command Syste Pini nsu Ustun eia Parameters nS Manual Bed_Detection O Valves End pa g Ona jemote_Contral Oko Select MasteiWizard a gt O Alams Master_Display Execute x ManFlow O Other End Fill G i Uain Result The adapter stops and the system closes the Bottom mobile phase
159. to prevent the column from draining If a packing test was chosen in the Method Wizard dialog it starts automatically Note Ending packing early or late will result in a too low or too high Packing Factor respectively 11 When the packing test is completed open the Evaluation module of UNICORN and perform an integration of the peak using the packed bed height value 5 4 Packing using the Master Wizard Description The Packing procedure described in the Master wizard guides the operator through a number of steps to pack the column with manual valves and a standalone pump Follow the instructions in the Intelligent Packing wizard found in the MAIN MENU in AxiChrom Master WARNING Ensure that nobody is present within the safety zone when the adapter is moving The operator in charge is responsible for keeping the safety zone free from unauthorized personnel and unnecessary equipment 120 AxiChrom 300 1000 columns User Manual 28 9562 89 AD Step 1 Select 5 Packing the column 5 4 Packing using the Master Wizard INTELLIGENT PACKING in the MAIN MENU AxiChrom Master packing wizard 1 15 a00 Welcome to the AxiChrom Master packing wizard This wizard will take you through a number of steps to confirm and edit your settings for the packing of your AxiChrom column Preconditions for packing are The systenvpump is primed All hoses leading to the column are primed The column is primed A Slurry tank
160. to 30 C 64 17 5 Ethanol 20 10 3h CIP 2 C to 30 C 64 17 5 acetic acid 64 19 7 Guanidinium 6M 5h CIP 2 C to 30 C 50 01 1 hydrochloride Hydrochloric 0 1M 1h CIP 2 C to 30 C 7647 01 0 acid pH 1 6 Isopropanol 30 1h CIP 2 C to 30 C 67 63 0 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 37 2 Specifications and characteristics 2 5 Chemical resistance Chemical Concen Time cycle Comments Operating tration restrictions temperature Phosphoric 5 8h For passivationof 2 C to 30 C 7664 38 2 acid stainless steel bed supports Sodium Oto3M 8 3h Purification CIP 2 C to 30 C 7647 14 5 chloride Sodium 1M 24 h room CIP 2 C to 30 C 1310 73 2 hydroxide pH 14 temp to 30 C Sodium 0 01M 12 months Storage 2 C to 30 C 1310 73 2 hydroxide pH 12 Sodium 1 M 20 3h CIP 2 C to 30 C 1310 73 hydroxide 2 64 17 5 ethanol Sodium 1M5 3h Adsorption 2 C to 30 C 7757 82 6 sulphate Urea 8 M3 5h Purification CIP 2 C to 30 C 57 13 6 Commonly 10 to 250 24h Equilibration ad 2 C to 30 C used aqueous mM sorption elution buffers for pH 2 to 10 chromato graphic use 38 1 When a concentration is given as a percentage this is v v 2 CAS no Registration number assigned by the Chemica Chemical Society FW concentrations than 20 can damage the column oN DM pH below 4 for stainless steel is not recommended Applies only to
161. to press the blue active CONTINUE button before the time bars have been filled This is to make it possible to speed up the specific process step but if nothing is done the procedure will continue as normal CONFIRM Opens the next dialog in the workflow after the stated prerequisites are fulfilled This button is active blue only when all mandatory in formation has been entered in the active dialog SAVE Saves the entered information and continues to the next dialog This button is active blue only when all mandatory information has been entered in the active dialog START Starts a process the adapter may move This button is active blue only when all mandatory information has been entered in the active dialog Stop Stops the process manually End Filling Ends the filling step manually and continues with the consolidation phase Active during the filling when it is grey If End Filling is used entered parameters might not be met and the packing might fail END PACKING Stops the adapter during the final compression of the bed END PACKING should be pressed when optimal Packing Factor or desired bed height within given range have been met This button is green when active PAUSE Stops the process temporarily Subsequent options are either to abort the process permanently or to continue Active when it is blue ABORT Aborts the current process and opens MAIN MENU Active when it is
162. tom of the column Carefully place a transparent ruler on the side of a clean and empty labscale column so that the zero mark on the ruler coincides with the surface of the bottom filter Make sure that there is a stopper at the bottom outlet of the column Secure the column in an upright position AxiChrom 300 1000 columns User Manual 28 9562 89 AD 91 4 Preparations before packing the column 4 3 Determining the slurry concentration 92 Sampling 5 Prepare the slurry in a tank of appropriate size 6 Make sure the gel slurry is homogenous The best way to achieve this is by stir ring Stir from all directions with varying tilt Allow at least 5 minutes for stirring This is the largest source of error in the method 7 Take a sample from the middle depth and width of the batch preferably using a Falcon 25 ml pipette Column filling 8 10 ii If the sample has an appreciably high concentration gt 60 dilute it by first adding 2 cm of water to the column The accuracy of the method is higher for lower concentrations Add thoroughly mixed gel slurry to the column until the right volume about the height of 100 mm is obtained Use a Falcon or Pasteur pipette 25 ml to avoid resin on the column wall above the 100 mm marking Add water until the glass body is filled Secure the end piece adapter and the filter on the top of the column Washing and settling steps 12 13 14 15 Connect
163. tone or salt solution to the column It is important that the column is properly equilibrated before the packing is evaluated 6 1 Choice of test samples for columns A solution of either acetone or salt solution for example NaCl can be used to give a good indication of the column packing quality The eluate is monitored by measuring the UV absorption at 280 nm or the conductivity The resulting elution profile is used to calculate the HETP value and asymmetry Both acetone and NaCl may interact with the matrix Do not use acetone together with the techniques HIC and RPC If using NaCl on charged media such as ion exchangers the bed must first be equilibrated with an eluent containing NaCl to a conductivity of about 35 40 mS cm before applying a higher concentration NaCl sample The sample is eluted by the equilibration eluent Consider the instructions for each specific media Alternatively the running buffer concentration can be increased 3 fold and used as sample solution provided that the running buffer contains salt to a concentration high enough that the sample does not interact with the matrix See Appendix for additional literature AxiChrom 300 1000 columns User Manual 28 9562 89 AD 141 6 Evaluating column performance 6 2 HETP and asymmetry calculation 6 2 HETP and asymmetry calculation General guidelines The sample volume should be approximately 1 to 2 of the total bed volume and 1 to 2 v v acetone can be used Al
164. um has been tested and verified by GE Healthcare AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 Packing the column 5 3 Packing using a UNICORN method 5 3 Packing using a UNICORN method Prerequisites This procedure assumes that slurry preparation and column priming have been performed according to the information on pages 90 and 93 The AKTAprocess system controls valves flows and rinsing procedures Note Ensure that the liquid levels in the containers are lower than the system outlet to avoid siphoning flow Note Ensure that the air trap is filled with mobile phase liquid This is because the methods created using the Method Wizard for priming are constructed so that the air trap is inline with the column when equilibration and testing operations are being performed The packing method is built to bypass the air trap Note During the packing procedure it is not possible to pause the process when the consolidation phase has started The packing procedure can be ended by pressing End in UNICORN or by using the Emergency stop If a packing proce dure is ended the procedure will be aborted and the medium has to be un packed from the column WARNING Ensure that nobody is present within the safety zone when the adapter is moving The operator in charge is responsible for keeping the safety zone free from unauthorized personnel and unnecessary equipment Running the method 1 Run the created packing method
165. umns User Manual 28 9562 89 AD 43 3 Getting started 3 2 Placing the column and Master 44 Zone Description A AxiChrom column Safety zone red B Service access area blue C Off limit zone yellow which is an area where cables and tubes are located D Approximate location orange of a connected AKTAprocess system option al Note that there will be more off limit zones with cables and tubing when an AKTAprocess system is connected Safety zone dimensions Column size mm 300 400 450 600 t f0 0 Safety zone length A mm 3200 3500 3800 Safety zone width B mm 2600 2700 2800 3100 3300 WARNING To avoid injury no persons are allowed to be present inside the safety zone when the column adapter is in motion during mainte nance procedures see figure on page 43 The operator in charge is responsible for keeping the safety zone free from unauthorized personnel and unnecessary equipment WARNING To avoid injury no persons are allowed to be present inside the yellow zone the area around cables and tubing running between the column to the Master see image on page 43 WARNING Move Master so that it is always close to where the work is per formed The EMERGENCY STOP button on Master should always be in reach AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 2 Placing the column and Master Moving the column without whee
166. wo tube lengths allowing maximum bed heights of 300 and 500 mm respectively The columns are validatable and the construction materials comply with the ASME BPE standards They are fully traceable back to the source for biopharmaceutical production All wetted polymer and elastomeric parts are classified according to USP Class VI 21 CFR Part 177 and are animal origin free or comply with the conditions in the CPMP Note for Guidance EMEA 410 01 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 5 1 Introduction 1 1 General introduction The images below show a 600 column left and an AxiChrom Master unit right 1 2 Scope of this User Manual This User Manual is designed as a general introduction and guide to using AxiChrom 300 1000 columns It is recommended that the user reads the complete Operating Instructions before using the column Information concerning safety precautions is presented in this manual in conjunction with general information about the features and operation of the column Specifically the instructions cover Chapter 1 General introduction intended use of the columns safety in structions and labeling Chapter 2 AxiChrom 300 1000 columns specifications including principles of column function and column specifications Chapter 3 Preparation of a column for use including unpacking installa tion column and system setup use of AxiChrom Master and instructions for cleaning the column before use 6
167. xiChrom 300 1000 columns User Manual 28 9562 89 AD 47 3 Getting started 3 3 Wheel kits Wheel kit components Part Function 1 Lever 2 Trolley 3 Trolley bar 4 Rear wheels 48 AxiChrom 300 1000 columns User Manual 28 9562 89 AD 3 Getting started 3 3 Wheel kits Wheel and trolley fittings 0 0 0 ic N i E a 90 110 mm WN TINANA NAGY NN A Hole for fitting rear wheels F Pins for locating trolley bar Note To be able to assemble or disassemble the wheels the distance between the floor and the column stand must be in the range 90 110 mm Trolley and trolley bar Part Function B Bolts C Height adjusting screw D Trolley knob E Slots 5 Safety pin AxiChrom 300 1000 columns User Manual 28 9562 89 AD 49 3 Getting started 3 3 Wheel kits Assembling the wheels on AxiChrom 300 600 columns 1 Place the lever close to one of the rear feet and lift the column by lowering the lever arm Screw the foot pads in completely f needed for work in restricted spaces the lever can be mirrored by changing the attachment of the lever arm 2 Fit the rear wheels in the hole in the column stand A Wheel and trolley fittings on page 49 Ensure that the washer and safety pin are corre
168. ystem The table below shows how to connect the column Master and an external system Numbers in parentheses refer to the illustration Connections on Master and column on page 59 CAUTION Use a pressure gauge pressure relief valve rupture disc or other pressure safety equipment to ensure that the maximum operating pressure of the column is not exceeded 60 AxiChrom 300 1000 columns User Manual 28 9562 89 AD From Pneumatic air supply 1 on the Master 3 Getting started 3 4 Connections To Wall socket air outlet 5 5 7 bar Motor power cable 4 on the Master Motor power the right connector on the column 13 Motor feedback 5 on the Master Motor feedback the left connector 13 on the column Pressure sensor connector 6 on the Master Pressure sensor mounted on system 14 Profibus signal cable 7 on the Master Profibus connection on an KTAprocess system Two air outlet connectors 8 on the Master Pneumatic inlets with 2 valves 15 on the column Slurry inlet 9 on the column Slurry tank Bottom mobile phase 10 on the column Bottom mobile phase on a system Col umn1 bottom valve on the AKTAprocess system Rinse inlet 11 on the column A system CIP2 Inlet on the AKTAprocess system Top mobile phase 12 on the column Mobile phase on a system CIP1 Inlet on the AKTAprocess system Protection ground cable on the column stand

AxiChrom™ 300-1000 columns - GE Healthcare Life Sciences

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AxiChrom&trade; 300-1000 columns - GE Healthcare Life Sciences

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AxiChrom™ 300-1000 columns - GE Healthcare Life Sciences