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Biacore T100 （Ver.1） 取扱説明書

Contents

1. CN hm Pe
2. a 1 Sensor Chip i CM5 Biacore T100 9 189 9 Evqluation Evqluation 9 1
3. 60 4 Z 2 a O O
4. OFF TEL 03 5331 9336 ED 9 00 17 30 WWW delifesclenceS COJD e mail Web
5. RI O RU RI RI RI 0 1
6. DMSO 1 1 500 RU DMSO DMSO 10 RU 3 100 RU 10 000 RU DMSO
7. 2 3 Baseline 132 0 5 3E5BB1E LO OD baseline 1 197 0 dE 5 1 38 E71 Binmdinn gg 5 Ed HH1E H I5 1 9 stability 1 ld baseline 1 10 binding _ 1 SI stability _1 10 NBGSMINe 1 RU 0 RU RelResp 0 0 binding_1 stability_1 RelResp baseline_1 RU 2 bagseline_2 binding_2 stability_2 RelResp baseline_2 RU Toolbar Reference line Menu bar View Reference Line C 9 Em a fi Ft
8. Biacore T100 6 167 6 4 Excel Excel Excel txt Next gt Next gt Biacore T100 168 6 6 3 E060417 conc Systenn Preparations x J Frime before run Normalize detector ETiDETatUTE settings nalysis temperature Sample compartment temperature 55 C Help lt Back Next
9. Include Next gt Kinetics Affinity Select Data Create Blank Subtracted Sensorqrams 0 0 0706 Affinity gt Biacore T100 5 67 Create RU Req RU Req Next gt Kinetics Affinity Fit Affinity Create Steady State Affinity No fits perfarmed Delete Req RU RE 2 2 9 Biacore T100 68 5 Kinetics Affinity Fit Affinity Create Steady State Affinity Steady State Affinity Finish
10. 2 2 AH AS 1 2 1 13 C 21 C EF 29 C 37 C 45 C nm kJ mol kJ mol kJ mol 4 Ko Ko kg
11. RU moulselq SL 3 TT Conc OOD ng ml Exclude Curwe Excludg Cycle RatinE ESE 15LiI ho Sensoregra Lahea le Cinny Granh Exnnrt urwes i 2nnn 2nnn amn ridlines 120nn Concentrat Legend ngi Exclude Cycle Calibration toble A Galibratian table 1247 0 3045 2490 2482 2 4529 0 4647 4 4 8 2 9999 0 3252 Parameters for curye 1 Rhi 2871E 04 Rlo 1471 Al 493GE 04 A2 1 na Biacore T100 5 83 5 21
12. 5 3 Menu gt Export Positions Menu gt Import Positions Biacore 1100 5 53 5 4 Control sqmple R1A1 R1A12 12
13. RU RU 40 x 1 S x RU 200 x 1 S x S 50 kDoa 100 kDa 1 40 x 1 1 x 50 000 100 000 20 RU 200 x 1 1 x 50 000 100 000 100 RU 20 100RU Biacore T100 3 23 3 1 N e CM NHS N NHS CONH COO CO CONHCH CH OH NFS
14. 1 1 4 1
15. 0 5 TFA 0 1 ZipTipTM Milipore 50 mM NaoOH NaCl Surfactant P20 DMSO Biacore T100 138 5 5 6 1 Toolbar Run Method Menu bar Run gt Method
16. Bulk_ Effect ng ml ug ml kKp 1 10 10
17. 1 1 Binding ng 11 Binding Biwalent nalkwte HasterngenEnuls nayte Hgterngehnnus LEand RU Tmo State Reaction confor mational charege Ft 5 9 B A 1 1 Binding 1 A B AB Bivalent Analyte 2 2 AB B 2 A B AB AB B BAB Heterogeneous Analyte 1 2 A1 B A1B A2 B A2B Heterogeneous Ligand 2 A B1 ABl A B2 AB2 Two state Reaction conformation chande 1
18. pH 3 Methods Manual run Biacore 1100 2 15 2 1 Toolbqr Start Manual run Menu bar Run gt Manual run 8 Phwpsht334 COCO 5 Flow rate 1 100ul min Flow path Rack Ca Start ul min s lu
19. End manual run Menu bar Commands End Run Standby flow 4 3 Tool bar Reference Line View gt Reference Line WE TEL View Biaqse Line
20. EDC 89 ul 7 mm NHS 99 NHS EDC 7 mm Ethqnolamine 128 ul 7 mm Ligand 98 ul 7 mm Biacore T100 34 3 Eject Rack Rack tray port Eject Rack Tray _ Hack Tray Ejected Click OK to return the rack tray to the sample compartment Hep Time to auto close 58 OK Eject Rack Tray Rack Positions Next gt WE Entrmhmltr mf HI irm Ft PTE il r EL hi fwe hm mmrg bater Meme hetmwss etn me mes tt Smt csm Start Sa
21. Insert Chip Dock Chip Biacore Ti Conktrol softwarEe nserting chip 1 2d Dock Standby flow Standby flow A 4 65 ml 24 1 5 Dock Insert Chip Cancel Toolbar Eject Eject Chip Insert Chip Toolbar Insert
22. Rmax 20RU SLEN CES Rmax x xS Da 50 000 Do 1 000 RU 20 000 Do Rmax 20 000 x 1 000 50 000 x 1 400 RU 10 000RU RU vs C k k Kp
23. Kp Ko k Biacore T100 100 5 vant Hoff Kp AG RT In Kp AG AH TAS InKp AH7 RT AS7R AG R T Kp AH AS SR intercent on Y aXis kJ moD K M kJ moD J K moD In Kn AHF R slope I T RT InKp AH TAS AC8 T To TAC5 In T To AC To kJ K moD 25C Eyring k k k ksT h exp AS R AH RT In k T AS R AH RT inkB h AG Biacore T100
24. Req vs C C Req K Kp Req Cx Rmax C Ko Req RU C M Kp 1 10 10 5 0 1 2 n 2 10 0 Biacore T100 5 49 5 1 1 Toolbar Run Wizard Menu bar Run gt Wizard Assay Kin
25. Ctrl Break Biacore T100 Evaluation Software 5 426 binding stability co bindind 1 stability CO_bindind Co_stability FTW 0VET control Redehneration Biacore T100 128 5
26. Aim for Immobilized level Toolbar Run Wizard Menu bar Run gt Wizard mr war Teete tf mix Surface Freparation i Methods AndTemplates Ilrmmobilization pH Scouting nmobilization ee Name Te 5 Fiegeneration Scouting Buffer Scouting Surface Performance Control Experimenls Kinelics Linked Fieactions Kinelics Mass Transfer Assay Concentration A amp nalysis Kinetics SFfinity 2 Thermodynamics Einding amp nalysis Help Browse Dpen Cancel Surface Preparation gt Immobilization New Methods qnd Templates Open Browse Open 5 inimobilization Immobilization Setup
27. Ko ka k k M 1s 1 k 1 Ka A B AB 0 Kp ka ka KA ka kg Kp kg kg R fl ka kg Rmax C ka k Biacore T100 48 5 fReqJ
28. Miscellaneous settings Detection 3 Single Dual Multi Single 1 2 3 4 Dual 1 2 3 4 2 1 4 3 Multi 1 2 3 4 2 1 4 3 2 1 3 1 4 1 Assay Steps OO BC Biacore T100 120 5 Startup Method Builder Main Overview 1 Create assay steps by using the buttons below Startup General Settings startup 2 Select an assay step by clicking on it Edit properties below or use buttons to remowe or mowe the step New Step Fiemowe Step kMoye Up kowe Down Copy Step Assay Steps 2 Sample sample Cycle Tupes Sample amp Assay Setup 3 Simulate Cycle Hun List 3 times as entered LMW screen LMW screen 1 time as entered Solyent Correction Salyen carrectian Before after every 20 Cycles Solvent correction 1 time as entered Control sample Control sample Before after every 20 cycles LM W screen 1 time as entered Base settings Startup Startup
29. BR 1000 50 EDC N ethyH N 3 dimethylqminopropylcarbodiimide hydrochloride NHS N hydroxysuccinimide 1 M ethanolamine hydrochloride pH 8 5 EDC NHS 10 ml 200 ul 7 mm 20 C 1 4 200 ul lt 1 BSA
30. Biacore T100 5 145 Eject Rack Rack tray port Hack Trau Ejected Click OK to return the rack tray to the sample compartment Hep Time to auto close 58 OK Eject Rack Tray Rack Positions Start a EH te a EE Fi ae th ww ml Start Save aqs Methods qnd Templates Bia Users Dont Save WE eu A Fs mmwr acoreeranmte Ge 8 Pe Imusr _ cm Save in File name Saqve Standby flow
31. Biacore T100 74 5 Ts Concentration Analysis System Preparations x Piime before run Narmalze detector Temperalure settings Analysis temperature 25 C Sample compartment temperature 25 C Help lt Back Nexl gt Close Prime Normalize Analysis temperature 25 C Sample compartment temperature 25 C Next gt LT rrlmuhhah Jr Fk Pol ee i ck 2 Fi 1 1 2 mW C 0 te emer Fiear mt sbrt ww eml ul 5 15
32. ul 1 14 Biacore T100 142 5 Conditioning ro tr pi Eb TT OLHPoL TH snmurrrriih Hoe hopic sy HH orm Bi Hm pnhuhomupedl Pr ma ELEI CPP rm hg hanarm le pa Fer i et tht ee I Care YWP Fr eye ei ET rit rile cr ed TFA rimm ed cn ast end Settings ly Regenaestion 1 Fregererstion shaor asETffa Loscl ime Fm yale mmm Eom pm sw Bmdp Hmh ricnsip ohn Ehawwhiwwmeetenmr ao Peas i Regeneration solution Contact time S Flow rate ulmin Flow path 1 2 3 4 Setup Run SE Method Builder Detection Detection Flow path 1234 1 Flow path 1 2 3 4 Next gt Next gt
33. temperature 25 Biacore T100 1 11 1 2 4 Toolbar Eject Rack Hack Trau Ejected Llick OF In return the rack Iran In the sample compartmert Hb Time to autn close HH Eject Rack Tray 60
34. Save as Methods qnd Templates Bia Users Dont Save FMe ammr necorrerwamtm Ge EE amusr Ce Save in File name Saqve Standby flow Biqcore T100 Evqluation Software Ee Biacore T100 5 77 5 17 Run gt Stop Run Biarnre Ti x 2 This will stop the run Help stop Run ar stop Run Fun Stopped FinishIn chrrernt cucle please Wal bort cnclg bn Lt Bre
35. 2 4 Saqmple Regeneration 1 2 Next gt Biacore T100 102 5 TE Thernnodynamics Setup Detection Flow path 3 5Startup Hun startup cycles Solution Dummy Number of cycles 3 Tempneralures nalysis temperatures Sample compartment temperature Temperature 5 Vary with analysis temperature Help lt Back Next gt Llase Detection Flow path Startup Solution Temperatures Analysis temperatures 2 1 4 3
36. Biacore 1100 5 81 RU mouselgG 4s00 35HI 2500 Relative response 15 In SHI 5I 2IH In 2 Un BI GU 10000 12000 Concentration nd 5 20 5 21 Concentration Analysis LCreate Samples VE ration Analysis Cres mouselgG 1083 0 4535 1129 1118 1129 1116 1123 D0 Retative rwPrmme a 0 1000 2000 XD 4000 HOD OD 7OO0 OO 000 10900 Measen ed concerir ation Finish Evaluation Explorer Sensnrgram nd All sensargrams Flot i Basgline Sample Be a Binding stability Cinntrnls stability Cnncentratinm nalysis Unncertratinn Analyss Evaluation Explorer Concentration Amalysis Biacore T100 82 5 5 20
37. 30 Biacore Maintenance Kit type 2 BIAdesorb solution 1 0 5 SDS BIAdesorb solution 2 50 mM Gly NaOH pH 9 5 Maintenance Tools gt Wash Buffer Tubing Start Wash Buffer Tubing x Select tubes to wash Al four tubes Next gt x This procedure remowes adsorbed material from the flow system The procedure is divided into three steps Total run hme is about 30 minutes NOTE se the Maintenance Chip for this procedure The surface on other sensor chips may be damaged by the solutions used Do not run this procedure below 20 C Do not abort this procedure after it is started eT Next gt uu Riequired salulions from Maintenance Kit BIdesarh salulign 1 ahaut 2H ml BIAdesarh solution 2 about 20 ml Next gt Biacore T100 wbourcrubo Step 1 Flace 20 ml BIAdesarh Solution 1 on the left hand tray and insert tube amp oe BlAdesorb Solution 1 Stqart
38. 10 mM 10 mM Borate 1 M NqC pH 8 5 Biacore T100 24 3 5 200 ug ml 10 mM pH 0 5 2 pH pH 3 5 25 Immobilization pH Scouting 10 mM pH SA4 100 ug ml 10 mM Borate 1 M NaCl pH 8 5 NHS pH 8 5
39. Biacore T100 6 151 General settings TE Method Builder Main 25 Buffer amp name The name of the buffer in position a BufferBname The name of the buffer in position B Buffer Cname UU The name of the buffer in position C BufferDname UU The name of the buffer in position D General settings 6 Concentration unit Data Collection rate 1 10 Hz 10Hz 1Hz Sample compartment temperature 4C lt 45 15C 10 DMSO Vary with qnalysis temperature Analysis temperature
40. Concentration Analysis 6 Concentration Analysis Toolbar Run Wizard Menu bar Run gt Wizard a Hae Bome Oean Crea Assay gt Concentration Analysis New Methods qnd Templates Open Browse Open SAMPLE Sample REGENERATION 1 Fiegeneration lt Bk New gt os 1 Capture
41. Startup 6 Binding to reference Stability baseline EE EE EE EL Biacore T100 5 137 5 6 2 ul
42. FTIVIC BiacoreT100 5 7 Sample Regenerdtion 1 2 Enhancement 2 Next gt Te Concentration Analysis Setup Detection Flow path 2 Startup IY Hun startup cycles Snlulian Ipumm Numbher of cucles no a Help lt Back Next gt Close Detection Flow path Startup Solution 3 Next gt TE Concentration Analysis Injectinn Parameters x Sample Contact time ln s Flow rate 5 MMmin Mi with Fractiar of the mis solution Regeneration Solution Im lp H Cl pH2 0 High wiscosity solution Contact time le ls Flo
43. Biacore T100 1 7 1 6 Reuse chip Newchip Heuse chip Fieuse chip Reuse Ehip tnpe Chip Chip Int hn hinck Chip Carnce Reuse id Details Chip lot no Firstuse date DED329 1151064 3429 2006 Chip IFC type CM5 IFC105 Close id id Biacore T100 8 1 1 7 Series S Sensor Chip C
44. Evaluation Biacore T100 Evaluation Software C Documents and Settings nakano y BIACORE1 My Deum en NERS g EE 4 File Wiew Evaluation Tools Window Help a Sensorgram Plot AConcentration Analysis A Kinetics Affinity A Affinity in Solution Thermodynamics Evaluation Explorer All sensorerams I Edit Remoye lt 4 hssay Sten Furpose Undefined 4 Cycle lt Overlay gt Tools Sensargram A sensorgrams Plot ga Baseline Sample 1 gq Binding level Sample 1 gq Binding stability Samp Binding to reference S Sensorgram zaaom Lock 5 29 Keyword table Tools Keyword Table Finish EE ayrwnrd Tablm THEE 1 ET Biacore T100 5 93 5 30 Menubar SNEO rram rdg mshilrfr Summs 1 113TmO Bdne t
45. Ctrl Break Biacore T100 Evaluation Software Biacore T100 78 5 5 2 2 Evaluation x File Wiew Evaluation Tools Window Help Sensorgram Plot Concentration Analysis A Kinetics Affinity Affinity in Solution Thermadynamics Evaluation Explorer All sensnrgrams Edit x Remove Curve Name Fc 4 3 4 hssay Step Purpose lt Overlay gt lt Cycle Overlay gt Tools Sensorgram Zoom Lack Sensargram ER All sensorgrams Plot i Baseline Sample i Binding stability gi Binding to reference Calibratian Calibratian2 Samhle Sample2 Startup Startup2 5 18 Keyword table Tools
46. Ru Um enrgrarm Een Lask 350 _ a a _ _ ae NN in mn EE hp m i _ 1 le En am 99 mo TlTig HH Ei Biacore T100 9 197 9 5 Excel Sensorgram window Copy Graph Biacore Word Pad Paint Word Pad EN 3 mlrlml some MEE EET EE EC EO EL EE TL
47. Edit EDC NHS 5nlutin EL ix with INHS an Fractior nf mi mith solutior Help Finntact time HEH tl Flow rate ho i OK hethod name ICopy f Amine nmmarid 5 lutinn Contact Time fs Flow Rate i ie 2 FRE CQ MC SpeciFied in Immnhilizatinn 5etun PRTMECT EDC MHS SO 50 Enn 1n jmjct Ethanolamine Lx LIGANDINIECT Specified in Immabilization Setup Mix amp mject a INIECT Ethannaming 2 1n EE Biacore 1100 3 33 TE Immobilization pH Scouting 5ystemn re Prime before run Normalize deteclor Temperature settings Analysis temperature 25 E Sample compartment temperature 25 Help lt Back Next gt Llose Prime Normgalize Temperdture settings Anaqlysis temperature 25 CC Sample compartment temperature 25 C Next gt Trmrnahilizr ation nee C 3 0 _ Jee ee ieeet Mk Nmz ge Rn ul
48. k KRmax lt Rmoax RU Rmax Rmax Rmax Biacore T100 5 63 5 11 4 Fi
49. Control sample R1A1 R1A12 12 Menu Automatic Positioning Si 2 Jr 2 I Cam de Em te Fi uch Bn Es a ri ri ir mmamn mrrh Tl ETP i sis EL bl a ITITEELW1 Lanhum itm Pep Jim EE i i mt nal 1 Pf 1 a Auto First reginn in the list is positinned first Rack Vial Size Sample Sample Sample gt Small gt Small gt Small Reagent _ Reagent ES Medium No Large gt autn Content Content Content Content gt Content Pooling Yes OK Automatic Positioning
50. Calibration Control sqmple Base settings Name Purpose Purpose Biacore T100 154 6 Evaluationsoftware 7 5 Calibration Conditioning Control sqmple Saqmple Solvent correctionStaqrtub Undefined Connect to cycle type Cycle types DUUST 2 Recurrence Calibration Control sqmple
51. Kinetics Affinity 47 Concentration Analysis 69 Binding Analysis 85 Thermodynamics 99 LMW screen LMW kinetics 115 Inject and recover 137 Biacore T100 5 47 5 1 Kp M 1 Kp KA 1 M A B AB Kp Al B IAB
52. Sensorgram window Export Curves txt Excel text Ero hi gs FTisirit imil ymi 5 IL H33 3 1 EE IMEI ll n i Eh I dd aki TA EF TE 178 0 1 185 9 1 1 L118 7 EE Ao IS1E T_TET T1054 57 sh TT 1 1 a 1 IET 1 it 3 1M 1 I 17 ll 5 a ICE EE 1 i Lh 1 NE Ii MI a 1 i 1 0 2 TH7ll 1 IPEEI I Ii 3 I 8 1 LE 6 INEigHE n 0 0269 3 1 a 1 fl 1 IRI E 95 TE I EEPM 19483 36 Ph Ti GE 7 1 4 0 1 Biacore T100 198 9 Excel File gt
53. pH Biacore T100 4 39 4 crude
54. Menu Export Positions Menu gt Import Positions Biacore 1100 5 75 ve Control sample R1A1 R1A12 12 Menu Automatic Positioning S OW Jr YY JJ I Cf Wm ote Fi eth nm Es mE ri rd ty RE Egon Fnlr ritrel tarps EC on mc a ET i EE tn lr rim Er ri Cri r EErHinmm ke HmsmoWHIt LT erinnt rrr mmm t rie 3 Auto First reginn in the
55. DMSO Instrument handbook Biacore T100 3 25 3 1 1 pH CM5 CM CM 0 5 pH pH 1Immobilization pH Scoutin
56. Inject RU 5 0 G Eo 6 163 6 3 Solvent correction Cycle types New 1 Create a new cycle tyne nr select an existing from the list Cycle types currently in method New SUN Dat
57. Menu Automatic Positioning Si OW Jr YY JJ I Cf Wm ote Fi eth nm Es mE ri rd ty gon Polor_ rtrd simrg IC on mm ET i tn lr rim Er T Crmnt Sr ng eri Lor 9 Auto First reginn in the list is nasitinned first gt Sample Smal Content gt Sample gt Small gt Auto content gt Sample gt Smal Content gt Reagent Medium NO _ ctent gt Reagent Large gt Auto lt Content Pooling9 Yes OK Automatic Positioning Biacore T100 54 5 Eject Rack Rack tray port Hack Trau Ejected lick OK to return the rack tray to the sample compartm
58. Start Save aqs Methods qnd Templates Bia Users Dont Save Save in File name Saqve il TE Fr Tar amp Md sRm LE eml Refesp EL BU WW EE 1 DD I EE Mg ri Hi sh Se FD mal tai Fd cg Tl Immobilization pH Scouting Standby flow Biacore T100 Evaluation Software pH Control Software Biacore T100 3 29 pH pH4 pH
59. 3 Define all variable values at run time Define all variable values in method Define some variable values in method and others at run time Biqcore 1100 166 6 Verification The method has been verified and can be used to set up a run Setup Run Te lethod Builder Detection Sample amp Assqy Setup Define all variable values at run time
60. 10 O0K Biacore T100 134 5 5 3 5 1 100 Evaluation gt Add Plot rc EE Plat Settings Flat name Plot Flat type Report Pomt ws Wariable C Report Paint ys Repart Paint Axis setting Y Axis XR Axis Report Point stability lt Wariable Cycle number lt Response Type Relative response Help Finish Cancel Plot name MW c
61. Nac 10 mM Gly HCI HCI Phosphoric acid Formic acid 10mM Gly NaOH NaOH Etnhanolamine Etnhanolamine HCI EDTA Surfactqnt P 20 Tween 20 Triton X 100 SDS Octylglucoside Acetonitrile DMSO Ethyleneglycol in HBS buffer Ethanol Formamide GuQanidine HCI Urea Biacore T100 pH lt 2M gt pH 1 5 lt 100 mM lt 100 mM lt 209 lt pH 12 lt 100 mM lt 100 mM lt 1M lt 0 35 M lt 5 6 lt 5 6 lt 0 5 lt 40 mM lt Z0 lt 8 lt 50 lt Z0 lt 40 lt 5M lt 8M 4 41 Toolbar Start Manual run Menu bar Run gt Manualrun lt Fi Pia Fowiam tnd OOO CO TY OOK Delechon n fom cellrt 13 ooo RT r mpm Fompah1 OU F W wews wmnn lt OO OO Ei Fowmpgh1 34 WI 1 oo OO wmr OOC CN ocO JI J KC I JC CO CO Be Eeetmuck sw pss Flow rate 30 ul min
62. 30 000 RU DMSO Biacore T100 5 117 5 41 Biqcore T100 Evaluation Software DMSO DMSO 1 x y lt DMSO 5 DMSO DMSO
63. 7mm 58 ul Flow path Detection Dual First 2 1 1 4 3 3 Second 2 1 2 4 3 4 Both 2 1 1 2 4 3 3 4 Detection Multi Mix with Fraction 20 20 80 Stabilizqtion period qafter mix Mix
64. nM ug ml 5 33 Excel Excel Excel tb Biacore T100 104 5 Next gt Te Thernmodynamics System Preparations Prime before run Narmalize detector emperature settings Analysis temperature RS Sample compartment temperature 5 Help lt Back Next gt Close Prime Normalize OO NO eo OO GE I 1 Me Cct uae gE Rit s Coes u 9
65. DMSO 1x PBS no DMSO 4 6 DMSO 49 DMSO 1x 9600 ul 100 DMSO 400 ul 10000 ul 6 DMSO 1x 9400 ul 100 DMSO 600 ul 10000 ul 4 6 DMSO 8 DMSO 4 DMSO 100 200 300 400 500 600 700 6 DMSO 700 600 500 400 300 200 100 700 700 700 700 700 700 700 700 ul Biacore T100 118 5 5 5 1 Toolbar Run Method Menu bar Run gt Method Biacore Methods CT EE TT PP 1 ET EH 1 lb Bmww I Shor eectie vieied nomenes Hm pm el LMW screen LMW kinetics Open LMW screen
66. 2009 GE GE 169 0073 3 25 1 TEL 03 5331 9336 FAX 03 5331 9370 e mail Tech JP ge com ISO 9001 2000
67. 9 A B AB AB Biacore T100 5 61 inetics 7 Affinity Fit Kinetics Create 1 1 Binding kal1 Ms kal1 s Rmax RU RIRU KplM Chi lt RUS Finish bulk effect Evaqluation Explorer Sensnrgram 9 rd fAll sensorgrams Plat i a Baseline Sample i Binding lewel Binding stabilitsy a a Binding to reterence Kinetics Affinity Lh Toolbar ytes 7 finiy VE ER Biacore T100 62 5 5 10
68. RI RI Ft Constant Initial value 0 Parameter Setting OK 9 Fit 5 12 Current Fits 1 11 Binding Tmo State Raactinn ntor mational che Delete Biacore T1000 Evaluation _ 9K evBy OK ESCHDtICh Current Fits Biacore T100 5 65 5 1 3 Evaluation Biacore T10U Evaluat Evaluation Explorer Edit Remoye ul a Lg lt assay Step Purpose Ov
69. 8 Custnm Methods I x Methods a Aldehyde New Copy IEIETE Te Surface thinl Method name Amine Command Solution Contact Time s Flow Rate min PRE CONC Specified in Immobilization Setup 5 IXINIECT EDC NHS 5O 50 420 10 PS WASsH Ethanolamine L Ei 0 LIGANDINJECT Specified in Immobilization Setup INJECT Ethanolamine 420 10 cwee jm Copy Biacore T100 32 3 Huslrmm Me thoes Ta bh LE ieee thm La Maskeimide LE Sr tne tr Ei Te at Te 5 Fee Fea pr 1 PRE Cofec Sp mh rrarestskaakion Setup i MnEeT EDE Me SS 0 in Fiect WAH Ethesrodarriree LIGANDINTECT Speed Irershiienkion Sat Ee ECT Fn Fl Wt Wels 1 Ok Methods Mgthnd name Iopyofamme nmmrmard 5nlutinn Contact Time 5 Flow Rate Timin Fre cnnc PRE CQI Specified in Immnhiligatinn setup TINIECT EC NH5 SO 1h Inject ha ELhannlaming LIGANDIN ECT Specified in Immnbilizatinn Setup Mix amp ect i a INTECT ELhannlaming 2 1n Wash
70. 3 25 5 Sample compartment temperatures Next gt Sample Contact time 20 ls Flow rate plemin Dissacialian time Em s Regeneration Solution ly 2 High viscosity solution Contact time s Flow rate min Stabilization period ln Help Sample Contact time Flow rate Dissociation time Biqacore 1100 lt Back Next gt Close 120 s 30 ul min 120 s 5 103 Regeneration Solution High viscosity solution Contact time 30s Flow rate 30 ul min Stabilization period Os Next gt Rumi gm Be _ Can Sumetes Bsk Mets em Sample id MW Dal Concentration
71. CM5 Dock HBS N Prime Menu bar Tools gt More Tools Test Tools gt System Check Start Select teSHS to tur This procedure should be run at 25 U wth a new 5ersor Chp Lt5 and with HB5 as runnmg buffer Choose Close if you need to change the sensor chip reset the temperature or change rurining butfer A 2 Fieagert pumps and blank injection I B Refractometer performance and flow cell jeakage I C inectons A Tests if the peristaltic pump is in order and that a sample injection with bufer from the Rieagert supply block is al night es System Check Next gt Buffer scoutind F x lf the buffer selector shall be tested test F put tube B C and D in water lf all inlet tubes are not intenhded to be used after System Check dont forget to ruh Empty Buffer Tubing after the test oe A HBS N buffer 10X B C D Buffer Scouting
72. LMW kinetics 5 47 Method Builder Main Overview 6 Biacore 1100 5 119 General settings Ts Method Builder Main General Settings 1 Concentration unit Data collection rate Sample compartment temperature Vary with analysis Assay Steps MM 1 25 temperature Cycle Tpes Miscellaneous settings After run Sample amp re settimg MS vale Descmpion 4 Speciy analysis escription temperature after uh BufferA amp name ThenameoFthebuiferin position A Thename of the buffer in position B IThename of the buffer in position C Setup Hun BufferDname Thename of the buffer in position D Detection 5 Dual 0 Concentration unit Data Collection rate 1Hz Sample compartment temperature 4 45C 25C After run
73. Evaluation Explorer Thermodynamics AS diss JZ Kmol PITBETKDRSI 07 Biacore T100 Sensargram ed All sensorgrams Plot ga Binding level Binding stability ga Binding to reference Thermodynamics BZ u globulin 5 115 5 5 DMSO nM uM Ko 1 10 10 5
74. Next gt Biacore T100 5 59 Blank Subtracted Sensorgrams lt Back afiniy gt ioKnetis gt Cancel 0 Kinetics gt 5 8 Remave Selection Undo Blank Subtracted Sensorgrams Help Back IL Affinity gt Ir Kinetics gt Cancel Remove Selection Biacore T100 60 5 Mode
75. Prime Biacore T100 10 1 1 2 3 Analysis temperature Menu bar Tools gt Set Temperature Tnnls Help Prime Shutdown Biacnrg T1HI Ewaluation TiHH Stop Standby Eject Rark we Fack Turmination InsETrh nhi Ejert hin et Temperature Preferences nrE Tnnls et Temperature halnsis temperature Samples compartimernt BBT ILTE LL Help Carnce 4 45 OkK 15 Stqtus bar temperature
76. RU mouselgG SI a TT innec 5OOD ng ml Exrlude Cute Exrlude ce RatiuE re T1550 ho BTIS FEF IT Laha lB nnw ranh Export GuFwEs 2nnn 2nnn nnn Gridlines 12nnn Contentrat Legend hdJITI Exclude Curve Calibration table RU mouselg Cury Cycle Ennr Reshnr1s CY B ITPrnmi erRUI ng yml wo RatiWE ESE ai 1 PTT LL PT a PT ET EE Ai PT LL 1 5 OOD OO ANN EOD SOO 1 13Tnn EE Biacore T100 84 5 5 22 Concentration Analysis Create Control Sqmple Add Add Custom Report Point x Report point Assay Step Purpose H My stahility2 ly Calibratinn Control
77. AbsResp Baseline RelResp Baseline RelResp Binnding level Evqluation Explorer Biacore T100 58 5 Toolbar Ki ts tfinity Kinetics Affinity Select Curves Create Fe or a eit Ta Po gS es tme C9 7 Show concentration series Show blanks Shaw ayerage blanks wb mpe mee OO omron Sample eKinetics Affinity Select Curves Create zosulfamide lc8S8 lfamide uro semid e _ Include Time S
78. Biqcore 1100 152 6 After run Miscellaneous settings Detection 3 Single Dual Multi Single 1 2 3 4 Dual 1 2 3 4 2 1 4 3 Multi 1 2 3 4 2 1 4 3 2 1 3 1 4 1 Biacore T100 6 153 Assay steps EE Method Builder Main 1 Create assaw stens by using the buttons belaw eneral Settings Startup Startun Concentration 10 times as entered 2 Select an assay step by clicking on it Edit properties below nr use buttans to Sample remowe nr mnye the step Cycle Tupes sample Concentration 1 time as entered New Step Calibration calibration Concentration 1 time as entered Before every 20 cycles RS Hemaye Step Werification Control Sample EE _ Meweup Contral sample
79. Biqcore T100 Contro Biqcore T100 Evaluation Biacore T100 H9 3 NN OU 6 DR 9 F OD HH
80. HBS PBS HBS EP 10X 1000 ml BR 1006 69 0 1 M HEPES 1 5 M NaCl 30 mM EDTA 0 5 v v Surfactant P 20 10 0 01 M HEPES 0 15 M NaCl 3 mM EDTA 0 05 Surfactant P 20 pH7 4 HBS P 10X 1000 ml BR 1006 71 0 1 M HEPES 1 5 M NaCl 0 5 v v Surfactqnt P 20 10 0 01 M HEPES 0 15 M NaCl 0 05 Surfactant P 20 pH7 4 HBS N 10X 1000 ml BR 1006 70 0 1 M HEPES 1 5 M NaCl 10 0 01 M HEPES 0 15 M NaqCl pH7 4 PBS 10X 1000 ml BR 1006 72 0 1 M phosphote buffer 27 mM KCI 1 37 M NaCl gt 10 59 Vv DMSO 0 01 M phosphate buffer 2 7 mM KCI 0 137 M NaCl 5 DMSO pH7 4 HBS EP 200 ml BR 1001 88 0 01M HEPES 0 15 M NaCl 3 mM EDTA 0 005 v v Surfqctqnt P 20 pH7 4 HBS P 200 ml BR 1003 68 0 01 M HEPES 0 15 M NaCl 0 005 Surfactant P 20 pH7 4 HBS N 200 ml BR 1003 69 0 01 M HEPES 0 15 M NaCl pH7 4 0 22 um Biacore T100 4 1 1 1 3 Start BIA
81. Open Browse Open a Fei nk en ri i ea Ei Cn be EE om eml 1 Immobape le cel1 Medl em am uss 1 1 Corlacthme FD Fowrwe Im mmm Fm mm Fr memoe law ce Mmsil me F bpd 6 6 6 F Ceridme HD rl Fes rae Im us al me cel 3 Immobace aw ci 1 Mewed i r Li Lnrihsri Frer Ri TE fw man Fi ce 4 lcs ow cr kehod Lged F rm ter il FEmm re li inbsmi Pe Hr Dard rit mm wm Chip type CM5 Flow cell 2 ly Immnhilige nw cell 2 hethod me im for immobilized lewel Ligand anti Beta2u glablin Hilule ligand Specify contact lima and ow rate LnmlackHimg 420 Flow rate fin ulermin Blank immobilization Flow cell Method Amine Ligand B
82. g CM NHS N NHS IC TT Ss S S 2 5 S Biacore T100 22 3
83. 4 HBS EP 50 ml 4 Sensor Chip a CM5 2 N 5 Sensor Chip CM5 Re Biacore T100 188 8
84. Keyword table Tools gt Keyword Table Finish er Reset All Filters buffer RE hdd Keyword Salvent correctian RE EC Sample Warfarin E Rename Keyword Wartarin 100 Warfarin 0 Remove Keyword Warfarin 25 Warfarin 125 Warfarin 625 Warfarin 3125 iWarfarin 15625 Wartarin 78125 Wartarin 125 i Naproxen i 0 Naproxen 100 i Naproxen 5 Naproxen 25 Naproxen 125 Naproxen 625 i Naproxen 3125 Naproxen 1 5625 Naproxen 078125 Napraxen 12 5 Furosemide Furasemide 20 Furosemide 10 Finish Cancel Biacore T100 5 7 Menubar 5 57 SNEO rram rdg mshilrfr Summs 1 Evaluation explorer Work areq Menubar Toolbar Evaluation Explorer ogram piot Baseline Binding level Binding stability Binding to reference Work aread 113TmO Bdne ts referemee Samem 1
85. Control sample definition Bun control samples Repeat control samplels every sample cucles Control samples positiwe control negative control Help lt Back Next gt Llose Biacore T100 5 73 Control sample definition Run control sample Repeat sample s every 20 Control samples Control sample id Expected conc Next gt Sample id Dilution factor 1 A 2 1 s3 1 gs6 1 as B i 2 H2 B 2 2 3 B 3 2 14 B 4 2 15 B 5 2 B e 2 dl Help Tip lt Back Next gt Close Sample id Dilution factor Next gt 5 14 Excel Excel Excel txt
86. Assay Steps Cycle types Assay Steps startup Samples Control Sqmples Cycle types Assay Steb Cycle Type Biacore T100 6 149 6 1 Toolbar Run Method Menu bar Run gt Method Methods qnd Templates Browse
87. Biacore T100 160 6 EA Extra wash after injection with Stabilization period s General Sample Dual Inject Dual Inject 1 2 Generqgl Capture AN Enhancement 2 Regeneration 40 High viscosity solution Carry over contro
88. Kp M Rmax RU RIIRU bulk effect ChiRU2 Evaqluation Explorer Toolbar Kinetics finity TN Biacore T100 5 69 5 2 Biacore B B 2 A RU A RU vs A A Slop
89. OK ede ee lgg bo Cre File Properties Wizard Templqate Method Method Wizard Results Sensorgram Current Cycle Range 900 Al celee Include event log for cycles Biacore T100 3 21 3 90 CM5 Biacore 2 N
90. 10 RU em Rarnk ig Finish First value Low Second value High Medium RU Binding level Zoom lock Startup Sample Yalue 10 Control sample Relative response binding alue 2 n 5 10 15 20 25 3D 35 40 45 Cycle number Biacore T100 96 5 Evaluation gt Add Report Point Table Evaluation Explorer Report Point Table Work area 600 7136 Biacore T100 Evaluation S MM Evaluation Explorer Sensargram 7 All sensorgrams Plot ig Baseline Sample ga Binding level Binding stability Binding to reference Carry over a Controls binding I Re ort Point
91. 1 5 1 2 1 2 1 Insert Chip T100 Mewchip Reuse chip Mew chip Chip tuDe LM5 Ehip id Lhin Int mo optional Dock Chip arc New chip Heuse chip New chip Chip tupe cs r EE Ehip id Chip lat no Ti 1 Series S CM5 Hglp Dock Chip Carncel ONew chip Reuse chip Chip type 7 Insert Chip x lt New chip Reuse chip New chip mewes ls Chip id 2006041 7 Chip lot no optional 151313 Help Dack Chp Cancel Chip id Chip lot no optional Biacore T100 6 1 E
92. Vial well position Contact time 60 gt 120 Injiect Cancel Eject rack tray Menu bor Commands Eject Rack Eject Rack Tray Click OK to return the rack tray to the Rack Tray Ejected sample compartment Hep Time to auto close 58 Inject command wialywgll position 1 B1 Lnntact hme lgd Lancgl Help Mimimum renulreHd ynlume in wialywgll Fnr this inigchnni 8 hl s OK Biacore T100 4 43 a rr Er oie Bi OD Co Td a ei iil i ee mak a ike Hi 4 Pr tr a Leck sca m Em ed FTOTE TI i mm Dre TCE er ta Th Pi t
93. 28ul C NBia Users 2 Save Biacore T100 16 2 1 FT Hr a 3 Biacore T100 2 17 2 1 1 Inject command Menu bar Commands Inject wialy
94. 2 SS 0 AS R In k intercept on Y axis J K moD kJ moD K kJ moD In k T AH R slope I T 5 101 5 4 1 Toolbar Run Wizard Menu bar Run gt Wizard LLPHTULmn mn WW I Assaqy Thermodynamics New Methods and Templates Open Browse Open Caplure AIYTPLE Sample REGENERATION 1 Riegeneration lt 1 Capture
95. 18 2 Eject Rack Tray x Rack Tray Ejected Click OK to return the rack tray to the sample compartment Hep Time to auto close 58 Inject command YY Contact tirme Cahnce Help imimum renulreH nlumg In ylal wgll nr this Imlectinh 8 hbl OK Eee Toman Bm ech bm EE a yr TT a Pe Iriect MA MN a Ingect tA OD ahHWeeg SO LD Spe efiomp lmrelmg 1 1 1 Fad i de mW Yem aar 1 1 PH J EE ED Fa bmrmri_ 1 1 RD HH 1 a Temperatuae EF SC RT resraasi un Srmpke comgartanent Lamprahmar neet 5 C et EC Biacore T100 2 19 2 1 2 RUU
96. 3 56 Next gt 5 24 Sample 4 2 3 1 REGENERATION 1 ETEE TIDI 1 Biacore T100 5 _ 87 Te Binding Analysis Injection Parameters Sample Contact time ln ls Flowrale ho min Dissociation time 30 s Regeneration Solution Gly HCI 2 5 High viscosity solution Contact time ls Flowrale 30 min Stabilization period p s SW Re Sample Contact time 30 s Flow rate 10 ul min Dissociation time 30 s Regeneration Solution High viscosity solution 40 Contact time 30 s Flow rate 30 ul min Stabilization period 0s Next gt
97. ug ml i NG 30 1 Biacore T100 40 4 4 1
98. PBS HEPES HEPES DMSO DMSO 5 DMSO DMSO 10 0 05 Surfactant P20 5 40 PBS 10X 1 x 1000 ml BR 1006 72 pH 0 1M phosphate buffer 27 mM KCI 1 37 M Nacl gt 10 DMSO 0 01 M phosphate buffer 2 7 mM KCI 0 137 M NaCl 5 DMSO pH7 4 DMSO DMSO 0 22um
99. 1 2 Step 2 ipe the tube with a moist tissue Flace 20 ml Bl amp desorb Solution 2 on the left hand tray and insert the tube BlAdesorb Solution 2 Start U 2 3 vshpurerubno li Step 3 Wipe the tube with a moist tissue Flace buifer or water on the left hand tray and insert the tube Start 3 Biacore T100 163 7 2 7 2 1 Normalize Biacore Maintenqnce Kit type 2 BIAnormalizing solution Menu bar Tools gt More Tools Maintenance
100. a Standby flow Biqcore T100 Evqluation Software Ee Biacore T100 6 171 6 7 Run gt Stop Run 2 This will stop the run Help stop Run ar stop Run Fun Stopped FinishIn chrrernt cucle please Wal bort cnclg bn Lt Bre Ctrl Break Biacore T100 Evaluation Software Biacore T100 2 7
101. Biacore T100 5 89 7 Control sqmple R1A1 R1A12 12 Menu Automatic Positioning S OW Jr YY JJ I Cf Wm ote Fi eth nm Es mE ri rd ty RE Egon Fnlr ritrel tarps EC on mc a ET i EE tn lr rim Er ri Cri r EErHinmm ke HmsmoWHIt LT erinnt rrr mmm t rie 3 Auto First reginn in the list is nasitinned first Rack Vial Size gt Sample Smal Content gt Sample gt Small gt Aauto content gt Sample gt Smal Content gt Reagent Medium NO _ ctent gt Reagent Large gt Auto lt Content Pooling9
102. Simulate Cycle Run List 1 r ee ar Ei Em LEE her lrmi mm Em Cheerer am lm ha Pm Li Recurrence Number of replicates Simulate Cycle Run List Assay Step Name Number Of Cyches Assay Step A ThermQ 1 iglmiag lsiulp Number of Cycles Assay step Staqrtup 1 Thermo 7 Control sqmple 2 Generate Cycle Run List a Ma Eh Na Nanar OF Cte A Hm ie Ar rhmn Fe RL timmmimmrtsmtm Cie he Fa 1 Tar Recurrence Number of replicates Biacore T100 158 6 Cycle types 8 Miethod Builder Main Create a new cucle type or select an exishin rom
103. Start 1 2 x Step 2 Flace a bottle containing 70 ethanol on the left hand plate and insert the four buffer inlet tubes A B C D 70 Start 2 3 moucrrunp rn Sten 3 Riemowe the tubes from the ethanal bottle and allow Ihem to hang in the air This step empties the buffer selector of liquid A B C D 70 Start Frpppurcrrboo The Empty Buffer Tubing procedure is completed BC D Biacore T100 19 7 1 4 Wash Buffer Tubing AB C D BSA
104. MiliO BIAdisinfectant Solution 30 ml Start U DEsnrh an Sanitize Huhhin Lesnrh and Saritize nreHure stEh 3 please wall Time left IE 3 4 Desorb and Sanitize teh 4 Mn the numh Ilet tuhes with a moist llESUE Flace water nh the left hand traw amd iert ump iflet tuhes Flaca water nn the right hand ran and insert the water inlst ube rk sg A B C D MiliO Start Hunninn Desorb amd 5anllize proacedure step 4 please mal Time lelt HG3 4 5 Biacore T100 6 9 Desorb and sanitizte EE x Step 5 Leawe tube tu In mater Let tuhes B C ard hi hann im the air Hrk sg A B C D Start
105. Menu bar Tools gt More Tools Maintenance Tools Sensor Chip Maintenance Menu bar Tools gt More Tools Tools 8 Tnnls BE Maintenance Tools Desarh and 5anitize Empty Buffer Tubina Mormalize wash Buffer Tubing Test Tnals 2 System Check 5erwie Tools How 5ystem Wash on Software Froblem Report I 5uperclean This nrnredure remowes adsorbed material Frnm the Flow swstem Total run time is ahnut 20 minutes hin nat run this proacediure below 20 C NOTE Lise the Mainternance Chip For this nrnredurg The surface nn nther sensnr chips ma be damaged hy the solukions used Last run Fime na Ep cw 2 Biacore T100 22 GO 9
106. Next gt TE Immobilization pH 5coutinq Systemm Pr Prime befare run Namalize detector Temperature settings Analysis temperature 25 Sample compartment temperature 5 Help lt Back Next gt Llase Prime Normalize Temperdture settings Anqlysis temperature 25 C Sample compartment temperature 25 C Next gt ul ul 93 Eject Rack Rack tray port Eject Rack Tray Tray Ejected Click OK to return the rack tray to the sample compartment Hep Time to auka close 58 OK Eject Rack Tray Rack Positions Start Biacore T100 286 6 46
107. Open Overview Biacore T100 150 6 6 2 sm 10 Cur lt wWkm Ume wtmmd CILr sion Cmcerrmiom me ss wrtwed Defore J averr 0 cydas Control Mmegle Cortra smote Cmcerstom 1 Ume as ertered Before everr 20 crdes MD Seie 1 wones by cx Ds 1 m Negenesmon 10md Gti p20 E0 po ports General Settings Verification 6 Over view General Settings Assay Steps Cycle Types Sample amp Assay Setup Verifivcation Over view Smee 1 ww by Cycle XO 15 Pegereigion Om GyHO p20 0 epor ports MM 9
108. 60 bound 0 recovered 4 5 8 mmz ng UEHBE ls ll baseline 1 4 4 bound MOUG ea8UHG 19Hg Biacore T100 148 6 6
109. Ee pn rs HI to ri FarnPMe re TT Co mmm er Nm lm mn 1 Tupe Lo re romumeon HethodVarishies Evawawam vambls ala li ries Irwel br himiocisntime Fm Ei mm Fm mi eoh Fredp Fe mt Frctiore DN of mn okution mee Eur rah fer mn Cm mr 5 Shalsm ferred Setting for Sqmple1 Contact time 60s Dissociation time 60s Flow path Both Extra wash aqfter injection with IE 0 50 DMSO Biacore T100 5 123 Setup Run Next gt Startup Sqmple Cntrol sqmple LIL Samplel Solution ConcluM MW Da Next gt 5 42 Excel Excel Excel
110. Move Up Move Down ItartuD 1 Startup Thermodynamics 5 times as Bhntered Thermn 1 5arnhlg Thermodynamics 1 time as erntered 5nlyert correction 1 Cahditinrin Solyent correction 1 time as entered Every 15 cycles Control sample 1 Control sample Thermodynamics 1 time as entered Every 15 cycles 30 Startup 1 Solvent correction 1 Control sample 1 Thermo 1 1 15 J Solvent correction 1 Control sample 1 Thermo 1 16 30 Biacore T100 6 157 6 2 Recurrence Number of replicates
111. Menu Export Positions Menu gt Import Positions Biacore 1100 5 _ 125 mr Control sqmple R1A1 R1A12 12 Menu Automatic Positioning S 4 NT HL I EO I TH Tee WE O 4 Se erg pli Ms Llz ST 7 EE Fr lrwmml Chm em ote Fi Frh nm Es mE Fr rd rate Th Fiirmmam Them Eel mm Hail ff Cotoe Frierntas Miam 3 mr i TT ck WRm ml Cr c EE tn lr rim Er ET ri Er i Fmt AG rm Fint Frercr ng mmm t rie
112. Sample id 1 Immpnri Conhol amples lt Back Next gt Llose Sample id Control Sqmples 20 Next gt 5 25 Excel Excel Excel tb Biacore T100 88 5 Ts Bindinq Analysis System Preparations Piime before run Narmalize detector Temperalure settings nalysis temperature 25 Sample compartment temperature 25 C Help lt Back Nexl gt Llose Prime Normalize Analysis temperature 25 CC Sample compartment temperature 25 C Next gt i 33 Positiv R1B1 i 33 A 1 Sa 5 R1 B2 i 33 A 2 Sample R1 B3 i 33 A 3 Sample R1 B4
113. seupBun Buffer D name The name of the buffer in position D Save Cycle Tupes _ Miscellaneous 5 Concentration unit Data Collection rate 1Hz Sample compartment temperature 4 gt 45C 25C After run OO OO BC Miscellaqneous settings Detection Multi 1 2 3 4 Assay Steps Biacore T100 140 5 Surface conditioning Number of replicates Times 3 Injection and Recover Number of replicates Times Cycle Types Biacore T100 Injection qnd Recover a Mh
114. 3 Auto First reginn in the list is nasitinned first Rack Vial Size gt Sample Smal Content gt Sample gt Small gt Auto content gt Sample gt Smal Content gt Reagent Medium NO _ ctent gt Reagent Large gt Auto lt Content Pooling9 Yes OK Automatic Positioning Biacore T100 126 5 Eject Rack Rack tray port Eject Rack Tray x Rack Tray Ejected Click OK to return the rack tray to the sample compartment Hep Time to auto close 58 OK Eject Rack Tray Rack Positions start a LL iD iri mn a te ve
115. AbsResp Binding level Bqseline RelResp Binding stqbility Baseline RelResp Binding to reference Binnding level Work area Evqluation Explorer Biacore T100 110 5 Kp kkg 5 1 Toolbar Kinetics 7Affinity Kinetics Affinity Select Curves Create nm Co__ mdde eydew sammle con Assaystp Fow mw BTama 28 IF Show concentratinn series Show blanks Show ayerage blankS _ Hep MeeRm OO agen Next gt Jata Create 2 Thermo Blank Subtracted Sensorqrams 0 Kinetics gt Biacore T100
116. 5 37 Keyword table Tools Keyword Table Finish Shh larlber Skate EiWer Se rmt tr EE ng Samele Pr Sam Wr Fir I5ample gWgr En Wr bar rs 1m Samnle Warfer 11 Fam NW i 1 names fr EE Ea 14 Sm Wh 15 Sa hrt carectien 1 ET TT 17 Sample spe tar EC i 19 Sampke En ET onemt rmerlipm 75amml Fu me Sumpke Furapemis ET Fruitn mae Biacore T100 5 109 5 38 Menubar SNEO rram rdg mshilrfr Summs 1 113TmO Bdne ts referemee Samem 1 Evaluation explorer Work areq Menubor Toolbor Evqluation Explorer ensoro ram i Bqseline
117. Desnrh and 5anitize 1 Hunninn Desnrh and Sartze nceHure step please wal Freparing Desorb and Sanitizte The Desob and Saritze rncEHure Is completed lawy the snstem to rh i starndby mnde nr at least 4 hnurs before hrfnrmin a rh Hark Llose 5 Standby flow 3 4 Prime 3 Close Biacore T100 180 7 7 1 3 Empty Buffer Tubing B C D Buffer scouting BC D 20 70 Maintenance Tools gt Empty Buffer Tubing Start Empty Buffer Tubing j Sten 1 Flace a bottle containing deionized water on the left hand nlate and insert the four buffer inlet tubes ee A B C D
118. 85 5 3 2 eeeesI 92 5 4 0 0 99 5 4 1 2 101 MM 108 5 5 ees 115 5 fl 118 eS 129 526 DN SN oi eee 137 SO DOI i ib 138 6 le 148 6 1 uuu uauuauuuiiaatsateatesaseaseaeesseeeaseeaseeeeeseaeeeaseeseaaeeaeans 149 82 150 63 7 168 Biacore 1100 a TA 172 TE ii i Sib 175 D0 71077 22 7 175 Ta DEO ON SN RRC lb 7 176 TSUEMDOIBUNe NUD Gk 180 LAN NED UE DN i GC GS 181 7 2 183 TNOMMAZE cc 183 Se Fa EPO TARO DI RON OO PRP OTDICD PRS NWN NP 184 8 LAINNNNN NNNN Nvv k h 186 681 OM ca 186 8 2 ee 186 6 3 VL DR 20 ee 187 9 kk 189 9 1 kk 189 G0 0 189 5 PR 191 9 3 1 LLL 4 d
119. Dock Standby flow Dock Prime Biacore T100 0 6 7 1 7 1 1 Desorb IFC 1 1 20 20 Sumple compartment 20 C Biacore Maintenance Kit type 2 BIAdesorb solution 1 0 5 SDS BIAdesorb solution 2 50 mM Gly NaOH pH 9 5 A Maintenance Tools gt Desorb Start pesorb x This procedure remowes adsorbed material from the ow sustEm Total run tirme about 2 minutes HOTE se the hairternahce Chip for this nrncEdure The surface or other sehsor chips may be damaged hu the solutions used Lin nnt un this rncedure belo 2 Hark Nast gt Next gt Required snlulinns from Maintenancg Fil desorb snlutinr 1 Bdesnrh snlulinn 2 Bark Llnsg Next gt BIAdesorb solution 1 BIAdesorb solution
120. Start Save as Methods qnd Templates Bia Users Dont Save ee Imaisr G Saqve in Par ES NN File name Sqve b 3 Standby flow Biacore T100 Evaluation Software Biacore T100 5 127 5 45 Run gt Stop Run Biarnre Ti x 2 This will stop the run Help stop Run ar Stop Run Fun Stopped FinishIn chrrernt cucle please Wal bort cnclg bn Lt Bre
121. Stqrt BIAprograms Biacore T100 Evaqluation Softwqre 9 2 File Open CBIA users C BIA users T100 demo file Binding Analysis OK aa 4 4 1 esy SImD Ptsw Uimwj Sensorgram Biacore T100 190 9 Evaluation explorer Work areq Menubar Toolbar Evaluation Explorer ogram jpiot Baseline Binding level Binding stability Binding to reference Work area Biacore T100 EO ram Sensorgram window AbsResp Baseline RelResp Baseline
122. bem I Sho tovotmite iredtmetne Bm pm owl Biacore Methods My fo lrewerewerwow ee ee coe Inject qnd recover Open Method Builder Main Overview 6 3E Suwface conditioning Condcnma Comvdkormo 3 kmes as enkered Tnject and Recover Symok jniet ond hecover 1 tme ssenbered SeWroi fot sitay step Suface corGhonno Temcetsuae 25 He lt Semngi lt cycie bDe orrionmg Regonvwolom1 15 TFA ls Repod ports Il Biacore T100 5 139 General settings Method Builder Main Aa General Settings 1 Concentration umlt Data collection rate Sampnle compartment temperature Wary with analysis ssau Steps ng 1 2 ls temperature fter run Sample amp A Assay Setup speciy anabss Setting vale Descmpion temperature after run Burfer amp name Running buffer The name of the bufer in position amp The name of he huifer in position B The name of the buffer in position
123. rr ds et E re i Biacore T100 20 2 Toolbar Add Report point Menu bar Edit gt Report point 9 Time hi eho sem i a a et mm Chk op He or gmwl ld 0 Baseline OK 2 1 3 End manual run Menu bar Commands End Run Standby flow 2 2 Menu bar File gt Save 2 3 File Print
124. 5 47 Tools gt Method Biqcore Methods LMW kinetics 5 5 1 LMW screen Cycle Types LMW kinetics mh Ruder Mam 1 Ha a ete cc ye OF EeeEt toe tg Pea hes let els mr Erien mit nn TS ir a Ee Or TPME hm itr ta et tH rs gd Crd CN hmrmrm bia Ey pri thm nm Carr rh ret hd meh Corre Ce REPwt FEmk Ce Stings er am 1 ees Oommend a Eh hr li wb Fimmmrmi Fm CE wad mm Meihod Wabler Eumuhon mines IT hii oti hm am ls Fiow le wm Flor pem lmeh rr Fc rksgh Fr or mv ol me gnmunm a Fr Epha wah te ee ht et NM lg pad lil Setting for Sample1 Type High performance Evaluation Valiables Evaluation purpose Kinetics Affinity
125. A Slop vs C B Calibration Curve A A A A A A A B B A ITILHLEIMIIII vary aeseesVe 1n 2D 4 SH 5 Tl HD a i100 110 Concentration LTETI Biacore T100 70 5 5 2 1
126. BR 1oo4 11 FF BR 1006 55 7mm Plastic Vials 1 5ml Plastic Vials 16mm Glass Vials 15mm Plastic Vials BR 100Z 1Z BR 1002 9 BR 100Z 09 ms BR 1006 54 Biacore T100 1 18 1 11 Reagent rack A B C 1 2 3 A1 A2 A BC D EF G Biacore T100 14 2 2 G Toolbar WI Ns Biacore Ti O0 Control software File Em Run Tnnls Help cae eosesses sna E 1 2 Manual run Application wizards
127. Kop 1 10 10 5 0 1 2 n 2 ee Maihnd Rinilder War ible ET TP T M Tim 1 snlmrimm Come tn IW Cea TE 1 HH kh hn1 E LH T Biacore T100 5 129 5 5 2 Evqluotion 5 1 ME3 lt cycle lt Overlay gt Fi ools lt Sensorgram I Zoom Lock ga Baseline Sample ag Binding level gg Binding stability fg Binding to reference gy Carry over ga Controls binding gy Controls stability Cortrol sarnple Sample Solyert correction Startup 5 48 Key
128. OK Biacore T100 12 1 1 10 Reagent rack Type 1 1 5 ml 11 mm x 20 Reagent rack Type 2 16 mm 4 mDx 9 15 mm 4 mD x 9 7 mm 0 8 ml x 24 Sample and reagent rack 16 mm 4 mx 9 15 mm 4 mD x 9 1 5 ml 11 mm x 24 7 mm 0 8 mx 45 Rack tray 96 well 384 well x 1 Rubber caps type 3 Rubber caps type 2 Rubber caps type 5 BR 1005 02
129. RelResp Binnding level Evqluation Explorer 9 191 9 3 Evaluation explorer Sensorgrqam All sensorgrams Work qrea Sensorgram window 9 3 1 Sensorgram window lcure Mami Fc r ssay Gteh Furpose Lndafingd wcrle Over ay r n curve Name Fcs4 3 D EA Ctrl Biacore T100 192 9 ce Ovary Fil Senaergram 1 ee i 4 ET i Undefined Batazrmicro
130. txD Next gt Biacore T100 124 5 Ts Binding Analysis Systenn Preparations x Prime before run Narmalize detector Temperature settings Analysis temperature 25 C Sample compartment temperature 25 E Help lt Back Next gt Close Prime Normalize Anqlysis temperature 25 0 Sample compartment temperature 25 C Next gt Mnihod Bidmr W HI ul 5 43
131. 5 34 Menu gt Export Positions Menu gt Import Positions Biacore T100 5 105 mr Control sqmple R1A1 R1A12 12 Menu Automatic Positioning S 4 NT HL I EO I TH Tee WE O 4 Se erg pli Ms Llz ST 7 EE Fr lrwmml Chm em ote Fi Frh nm Es mE Fr rd rate Th Fiirmmam Them Eel mm
132. B CD Next gt Biacore T100 lt 195 ov ue emem ez sw owl BIAtest Solution 1 5 ml 695 ul 1 5 m 4 Start 8 asu lt Crcm File name Save BC D Empty Buffer Tubing 180 System Check Date p Biacore T100 Control Softwyare Version 1 0 id 12113 Configuration IFC105 05 Sep 2005 Evaluation Date 05 Sep 2005 0 os50905 Z086 Z600to 1400 RU OK 6 50to 50 RU OK B Refractometer atest solution Baseline level 22715 21400t0 23600 RL OK 35922 lt 2691 z140nto 23600 RU OK 35427 22514 21400to 23600 RU OK 35574 22550 21400to 23600 RU OK 3S6032 Average 22618 35739 Yariation 201 600 RU OK 605 lt 3000 RU OK C Injections Rise Fall Fc1 1
133. Biacore Maintenance Kit type 2 BR 1006 51 BIAdesorb solution 1 95 ml x 2 BIAdesorb solution 2 95 ml x 2 BlAtest solution 65 ml BlAdisinfectant solution conc 10 mlx 3 BIAnormadlizing solution 90 ml HBS N Duffer 10X50 ml Sensor Chip Maqintenance 1 BIAdesorb solution 1 4 BIAdesorb solution 1 4 Biacore T100 174 7 7 1 Toolbar Eject Menu bar Tools Eject Chip A This will eject the sensnr chip Help Eject Chip arirl Eject Chip Sensor Chip Maintenance New chip Hause chip ew chip Chip tunpe EE nn Chip id Ehip Int no Innhnnal Help hnck Chip Lancml Insert Chip Chip type Maintenance Chip id Dock Chip
134. NHS NHS pH8 5 pH pH pH5 Immobilization pH Scouting RU 100ug ml Immobilization pH Scouting Biacore T100 S007 3 1 2 Toolbar Run Wizard Menu bar Run gt Wizard lmoEsimomi ET Eau Surface Preparation gt Immobilization New Methods qnd Templates
135. cycle occurrences everly I Furnassaysteponce last Repeat assay step within Every Distribute I Hun assay slen nce irst Number of replicates times lt As entered 1 2 3 1 2 3 Order 11 2 2 3 3 Handom Assay step preparations Temperakure Buffer Number of replicates Times Biacore T100 5 _ 121 Solvent correction Uw sereen HUmewrenhered Before Jafber favery Hh evess h mw Recurrence Cycles Number of replicates Times M Control sqmple Satu fun me gm le Solvent correction Biacore T100 122 5 Cycle Types Solvent correction Ta Era it Ha Cal CHeC mnmi EF Cortmia mWetihond ni et ener ra hm lm Shred adimcfin 1 The Goh eet Ce Ptr nl ee Par hal ear mmih g IlGwm rmlm 3 i Command LMW screen
136. Target Reached Response os 3 5 NHS ii Chip CM5 Response Response Flow cell Procedure NMethod Liqand Bound RU Final RU Tarqet Reached Target level Amine proteina No Preconcentration binding is too Fast Help Frint Preconcentration binding is too fast pH Preconcentration binding is too slow
137. X Adjustment Report point time 0 boseline Biacore T100 9 194 Y Adjustment Report point response 0 baseline mm 1 mi RamwcPa fm gl Er ao kre T 1 1 i IN LE FM 1 ae MI m Wi i EE TimmE et 9 3 4 dt semergram il Tac 1 1 i MD 8 g lan 1 nm 3 Ti TD TE I Feom Leck Sensorgram window Cut hmIE1tm en gram Lil A a se j 130 1 0 4 s MU E 3 Um mr F gm 1 l n
138. ILMw screen Name Purpose Connect to cycle type Riecurrence cycle occurrences everly Hun as say SIen niCE last Repeat assaw step within Eyerp Distibute Hun assay Sten nriCE hrst Number of replicates B times lt As entered 1 2 3 1 2 3 nrder 1 1 22 33 Random Assay step preparations Temperakure Buffer Number of replicates Times Samples Method Builder Main Overview 1 Create assay steps by using the buttons below Startup General Settings startup 2 Select an assay step by clicking on it Edit properties below or use buttons to remowe or maye the step New Step Fiemowe Step Mowe Up Move Down Copy Step Assay Steps 2 Sample Cycle Types sample Sample amp Assay Setup Werification Simulate Cycle Fiun List 3 3 times as entered LMW screen LMW screen 1 time as entered Solyent correction Salyen carrectian Before after every 20 cycles Solvent correction 1 time as entered Control sample Control sample Before after every 20 cycles LMW screen 1 time as entered Base settings sample Sample 552 ILMw screen Name Purpose Connect to cycle type Fiecurrence Sample
139. Methods and Templates 0pen Browse Open 8 He ei Des Flow path Next gt TE Immobilization pH Scouting Injection P 1 Ligand Solution ant Beta2u globlin Contact time len sl Flow rate fo plemin Suface regqeneration This surface wash will be run once at the end of each cycle Solutior 50mt NaHH Help lt Back Next gt Llase Ligand Solution Contact time S 60 s Flow rate ul min 10ulmin Surface regeneration Solution 50 mM NaOH Biacore T100 S 7
140. 181 Evaluation gt Add Solvent Correction nnlrent Correction Zoom Inc 4D 2HH OH 2 HH GD SD 100D 1200 Response Ref FRL OK Evaluation Explorer Sensnrgram Pd ll sensoregrams Plat 6 Baseline Sample 3 Binding lewel 9 a Binding stability Binding tn reference i Carry ower 6 Cnntrn hindimg A i Controls stability Snlvent Cinrrection 0 Ja Snlyent nrrectinn Biacore T100 132 5 5 50 RU Zoom loc Copy Graph Export Curyes BHH BOD 400 200 0D 200 4O0 BOO0 BU 1000 1200 Response Ref RU Exclude point RU zoom loc Response Act Ref 15 BHH
141. Biacore T100 146 5 5 55 Run gt Stop Run 4 A This will stop the run Help Stop Run ar stop Run Run stopped FinishIn currEht cucle nlease Wal hnrt cnclg bn Lt Brg Ctrl Break Biacore T100 Evqluation Softwqre Biacore T100 5 147 a Ru bzx HN d 4 pd p ro 2 lt ll HIIH EOD 3 rHIIH BIIHI 5HIIIH HIH 33HIIH 2HIH 1 2 3 HI HI SH BI MI HI HH Time 1000RU 1 ng mm
142. 5 111 inetics Affinity Fit Kinetics Create 11 Binding No fits performed F le 6 4 nM ey LU Finish Evqluqtion Explorer 5 39 Biacore T100 112 5 5 39 Evaluation Explorer MEMS LU Backspace ah kinetics 7 ffinity Toolbar Fmeties Affinity Kinetics Affinity Select Curves Create Urves Sample Bz u
143. Hail ff Cotoe Frierntas Miam 3 mr i TT ck WRm ml Cr c EE tn lr rim Er ET ri Er i Fmt AG rm Fint Frercr ng mmm t rie 3 Auto First reginn in the list is nasitinned first Rack Vial Size gt Sample Smal Content gt Sample gt Small gt Auto content gt Sample gt Smal Content gt Reagent Medium NO _ ctent gt Reagent Large gt Auto lt Content Pooling9 Yes OK Automatic Positioning Biacore T100 106 5 Eject Rack Rack tray port Hack Trau Ejected Click OK to return the rack trau to the sample compartment Hep Time to auto close 58
144. Target level RU Washsolution 50 mM NaOH Next gt 1 30 35 lt EE Fi cr TE Cr Gal bres Ilmrrmmm 4 ata ul lauEllm mm CE 3 rim eae Fn 1 LT 19 ns Ferre Lh FT E 5 1 1 eCrrr a 5 MHD TE Ear Ta E Fe 1 mameLrm TA a 5 MWT1 EE mr PF TT 1 TEE He ET HS 5 TH OT RE Ho 1 2 MT EP Pdees 1 LT 5 mA 1 Mm mM MA Ha Fen 2 EE 5 ET a EE OS ET Ha Fn 13 EE Ne Fdset 1 1 DE DE ES 5 Lo TE Lu11 ld LS TT He neg nr Em Team eS tC ter hp CS Sumpls cpmpartmaiTe temperahe Gment 5 C et Biacore T100 38 3 riethod Liqand Boun Final anti Beta2u lohlin 1215 8 1389 7 Immobilization Results RU
145. Tools gt Eject Chip x A This will eject the sensor chip Help Cancel Eject Chip Chip type Maintenance Chip id Maintenance Dock Chip Dock Standby flow Menu bar Tools Prime Prime Biqcore T100 control software Biacore T100 Biacore T100 8 187 8 3 NO 2
146. x gt Chiptype gE Flaw cell 1 HOGE eho Amine im fa imrnBilized ey Ligand Diiute liqard GECifuEnritact iiE ri ii ratE Contact time 420 s Flow rate MLmin BITKimrinhilizgatigr Flow cell 2 i He A ine Ti of TITabized IEyEl Ligand ES iinte Ianid Secifncorntact iiE grid ig ratE Contact time an s Flow rate ho Mrmin BITKIimmiuhilizligr Flow cell 3 Immahilize flow cell 3 kiethod Amine 4 Eiri T IipHIIIgEH IEwl Ligand EE jntE II aid f 5nECifp corntact iiiE Tid ig ratE Contact time an s Flow rate ho MMmin irinhilizaliaT Flow cell 4 Immahilize flow cell 4 kiethod mine a i fT Tobized eye Ligand gE inte liqarig Contact lime zo s Flow rate fn ulmin Geciincontactlinmearnd W TE Kimmrinhilizgalia Help Custom klethods lt Back Llase Biacore T100 3 37 Flow cell 2 ly Immnhillze Fo cell 2 dethod mhe Lim For immnhilized lewel Liqarid arti Beta2u glablin hilute lgand Specify cantact Hm and ow rate Target lewel In HU ash solutior lg mk NaHH Bank immnbilizalian Flow cell Aim for immobilized level Method Amine Ligand
147. 5 1 1 1 n n 150 300 wm mn Pin Time 0 harwefimwet 5 Biacore T100 9 195 9 4 Sensorgram window Scqle Auto Min Max __9K_ 3 Biacore T100 196 9 Legend Right Hidden __9K_ Ra djusted sensorgram Som 9 te i Be 3 i HM gi Gridlines Moajor Gridlines Minor Gridlines 9K
148. Startup 1 Startu Thermodynarmics tirmes as entered Enhtrnl sample 1 Cnrtrnl sample Thermodynamics 1 time as entered Thermo 1 Sarnhlg Thgrrnndyharmirs 1 time as trg Stoartup 1 Control sample 1 Thermo 1 Recurrence Control sample Control sample 1 RiEcUrrEICE lw Repeat assaw tep mithin ITherma 1 Eyery 15 hermo Distribute Pun assay step ohce first Thermo 3 Repeat assay step within Thermo 1 5tartup 1 Startuh Thermodynamics 5 times a5
149. 0 Biacore T100 4 45 4 4 Application Wizard Assay Development Regeneration Scouting 5 Te Reqerieration Scouting Experimental Pararneters Hegenerahon parameters Stablicahon pernod lp sj ERegeneration Scouting Results 6 x Trend Chart Sensorgrams Sample Response Baseline RU RU 740 47450 Number of cycles for each conditior 5 lt 730 i 1 1 720 Settings AM 47350 Condition Regeneration solution Contact time fs 8 7m0 mm 4 EE B90 47300 8 EE CETTE 5 8 E80 8 2 Glycine 2 jpcme pH 2 25 so 4725n 3 Glycine Blucine pH 2 0 BEn 47200 i IElc
150. Add Custom Report Point Repart paint Assay Step Purpose stahility2 Calibratinn Cantral Sample Window 5 gt Startup Pasition the report li5 seconds atter lt Sample 1 injection stop lt Calculate respanse relative to report pa lt Help OK Cancel OK nase mm Rahrarl Paimtg Biacore T100 5 99 5 4 Biacore AG AH AS AG AH A S
151. Flow path Reference subtraction 2 1 4 3 2 1 3 1 4 1 Rack Start Pmpme Smwmmbme Fant Rie Tbh Ceal TE FE Te ir hg DE LP Fe RF Maske bd iT ea Bidcore T100 42 4 4 2 View gt Show Only Current Curve 1 yv View gt Show All Curves NC EC View gt Show Curves of Same Type 05 0 6 Inject command Menu bar Commands Inject 3 wialywgll positior 1 B1 Lnntact hme lgd Lancgl Help imimum renulreH wnlume in wialywagll Fnr his injgchnni 8 hl
152. Solvent correction 6 1 Number of replicates As Entered Order Random Assqy step preparations 1 A Biacore T100 6 155 6 1
153. A This wl stop the run Help stop Run ar stop Run Fun Stopped FinishIn chrrernt cucle please Wal bort cnclg bn Lt Bre Ctrl Break Biacore T100 Evaluation Software Biacore T100 56 5 5 1 2 Evaluotion Bia Templates demodata Kinetics Affinity 1 1 interaction blr Evaluation Explorer i Al senso Eat Oem El Sensorgram Zoom Lock ga Baseline Sample Binding lewel Binding stability ag Binding to reference Sainple Startup 5 6
154. Concentration 1 time as entered Before every 20 cycles Maye Down Copy Step Simulate Cycle Hun List 2 ase settings 3 ecurrence i Startup Repeat assay step within EErp cycle Purpose Startup DiSHiBule tocurences evenly Connect to cycle type Concentration FR assan slEn nrice First Hu assau slEn picg ast L umber of renlicates U ssay step preparations IO times Temperature As entered 1 2 3 1 2 3 Bulfer Order 1 1 2 2 3 3 Random Assay steps 5 Assay step New step Purpose Cycle type 6 3 Move Up Move Down
155. DMSO UV spectrometry DMSO pH DMSO pH Biacore T100 116 5 SPR RII 100 RU
156. MiiQW RE a i 4 Biacore T100 2 1 1 1 2 A 2 r Milli0 MiliO8K 1 2 ABCD 2 BCD 500 ml 2 Biacore T100 1 3 1 3
157. Total run kine is about ne hour nllnwued by a recommernded standby kine of hours HOTE se the hairnternahce Chip for this nrncedure The surface or other sehsor chips may be damaged by the solutions used Do nant un this nrncedurs belo 20 L hin nnt ahnrt this nrnceHure after it is startH Hank Mast gt Next gt LEsnFH and sanitize 3 x Riequired snlulinns Ifrnm hdaintenahnce Fil IdHEsnrh snluhnn 1 bout 4 ml Bl desorb snluhign 2 about 4 ml BdisinFeclant solukiorn ahnut BO ml ack Llnsg Next gt Biacore T100 5 2 7 Diesnrh and Sanitize 3 3 x Step 1 Flace 25 ml Idesnrh 5nluhinn 1 nm the left hard Iran and insert pump Ilet tubes Flace 15 ml IAdesnrh 5nlulinn 1 nn the right hand tra ard insert the water inlet tube lt Back poe BIAdesorb Solution 1 25 ml 15 ml 2 A B C D BIAdesorb Solution 1 25 ml MiliO BIAdesorb Solution 1 15 m Stqrt Desorb and sanitize Hunninn Desorb and Sarnitize procedure step 1 please wal Tirme left D D 4
158. Yes OK Automatic Positioning Biacore T100 90 5 Eject Rack Rack tray port Eject Rack Tray lt x Click OK to return the rack tray to the Rack Tray Ejected sample compartment Hb Time to aula clase 58 OK Eject Rack Tray Rack Positions Start ni EE EFT Erapars Fiun Protosoad ra bafta 1 i ea Ha Fi nn sem EE EE rw mm pem Start Save as Methods qnd Templates Bia Users Dont Save ET E
159. 1 2 Liesnrh and Sanitize x Step 2 pe the purmp inlet tubes wlh a moist lissue Flace 25 ml Bladesorb Solution 2 on the left hand tran and insert pump inlet tubes Flace 15 ml IAdesnrh 5nlulinn 2 nm the right hand tray ard insert the water inlet tube ga ge BIAdesorb Solution 2 25 ml 15 ml 2 A B C D BIAdesorb Solution 2 25 ml MiliO BIAdesorb Solution 2 15 ml Start Desorb and sanitize Hunmnnn Desorb arnd Sanitize hrnceHure stlep 2 please mal Time le OE A 2 3 Biacore T100 0 TL Diesnrh and Sanitize 3 x 5teh 3 Myine the purmp ill tuhes with a moist tlEsUE Flace 5 ml dluted Badisintectant 5nlutinn or the left hard tran amd insert purmp Inlst tubes Flaces dl ml dluted Badisintectant 5nlulinn nm the right hand ran and Insrt he water inlet tuhe lt BIAdisinfectaqnt Solution 50 ml 30 ml 2 A B C D BIAdisinfectant Solution 50 ml
160. 191 232 RD DR Gi 193 9 323 193 9 3 4 ee 194 08 195 7 197 CRE EE 199 Biacore 1100 1 1 1 1 1 1 1 1 gt gt gt gt Windows biacore LED reqdy temperature 1 1 Sqmple compartment door Sensor chip port Saqmple compartment injection window Rack tray port
161. 2 16 mm 15 mm 1650 Start Desorb Standby flow 3 4 Prime 3 Biacore T100 MO 2 7 1 2 Desorb and Sanitize 1 1 1 20 C 20 25 0 AB CO DO O AM IO Biacore Maintenance Kit type 2 BIAdesorb solution 1 0 5 SDS BIAdesorb solution 2 50 mM Gly NaOH pH 9 5 BIAdisinfectantsolutlon 6 ml 80 ml Maintenance Tools gt Desorb and Sanitize Start Desorb and sanitize This nrncedurg remowes adsorbed material and disintects the nw sustem The nrncedurg is divided into fee steps
162. Biacore T100 5 143 Te Bindinq Analysis Systemn Preparations Fiime befare run Narmalize detector Temperalure settings nalysis termperature 25 Sample compartment temperature 25 Help lt Back Next gt Llose Prime Normalize Temperdture settings Analysis temperature 25 Sample compartment temperature 25 C Next gt Mathead Huilkde k Fasm Yale WW OR OO a IT rmiM MHC iC KO 1 De Hr Ftd Er ei Ca FF Eg rl TF JK 1 OOO OL COW CX OO OOO 2 ea OK OO BOO KO OK 22 OO KT e009 ul Biacore T100 144 5 5 54
163. Bu niler Ma 5 141 lrrhmri Eh rip mod car 3 Iimihghmr etre lm Bsreh perird ammwrdt tepow Pot lewe A lst Pormmm emmmyal Fipm Ciece IT1 em Dc ECUMgWtecmraf 1 Sample solution Contact time Flow rate Flow path Wash solution Recovery solution Incubation time Deposition solution Deposition solution volume Number of repetitions Sr hnhon aagm Coniact fie EN Fler rime ten Fm Catt Fs3s Wt ONE TFA HgcryreTy Wumr Tf Flon bens PeFoawion schutionc wamrHEG Deposkion sionvekme 0 i Fun of eget s ulmin 1 2 3 4 0 Incubation time s
164. Sample Window fB lt 2 Startup Pasition the report is seconds atter lt Sample 1 injection stop gt Calculate response relative to report pal lt Help ok Cancel OK Case Rahrarl Paimtg Biacore T100 5 85 5 3 5 3 1 Toolbar Run Wizard Menu bar Run gt Wizard Hp Blowese New Upmn rmenml Assay gt Binding Analysis New Methods and Templates Open Browse Open Caplure Enhance
165. Table A Solvent Correction Fc Report Point Assay Step ware C Bia Users Templates demodata KineticsAffinity small maleculesblr Report Point Table 2 co baseline binding Sample PlResp RM 2 9 Biacore T100 Baseline CycleType LMWW kinetics Lid kinetics LMW kinetics LMWW kinetics Lhdw kinetics LMWP kinetics 5 97 5 31 Evaluation Explorer Plot Controls binding Biacare T1UH Eyaluatian Saftware C Bia Users Templates demodata Kinetics Affinity small maleculeshlr Controls bindineg Plet entra Evaluation Explarer lt lt Curve Name Fc 2 1 lt Cycle number lt Overlay gt x Controls binding I Zoom Lock Sensargram All sensargrams Plot gg Baseline Sample Binding
166. TharmodrmrTKs 1 me ss enteared 3 1 We 35 entered 1 me Ss Uared Befoee after J ewery 15 crdes Before after every 15 Cycles EL IAssay step 1 Conditiorirg Not cormected 1 me entered we gt Bose tetines Recurence Hame si 1 Popeal ssssy stmD milhm Purpose onditongwe 4 df cccurrences erer Comwct19 Cycle bpe bt Corrmmc ted lt F Fr Assay step 1 Assay step 1 Base settings Base settings IEalyent COorrection Mame Salwent Correction Furpose Connect to lt Cycle tn nt Conrnected Mnt Cornnected Ther mod hamics F Number nf replic nlyet correction a Name Solvent correction Purpose Solvent correction Connect to cycle type Solvent correction Recurrence Number of replicates Assay step preparations Biacore T100 6 165 Saqmple amp Assay Setup Sample Calibration Control Sample
167. gt Llase Prime Normalize Next gt ul MR eo Menu gt Export Positions Menu gt Import Positions Eject Rack Rack tray port Eject Rack Tray x Hack Tray Ejected Click OK to return the rack tray to the sample compartment Hep Time to auto close 58 OK Eject Rack
168. i 33 1A 4 Sample A A 1 R1 B5 i 331A 5 Sample hd swe MNO Ul OOO R1 B12 33iC 2 Sample CO OCOCOCOOO SE SS 000060060 EE 0 8 R1 D1 79 Buffer Startup OCOD R2 A1 1340 Gly HCI 2 5 Re 1 1 7 1 1 7 OOCOCOCO 0 OOOCOOO s0O OOO OOOOO e OOOO Help Menu Eject Rack lt Back Next gt Close ul WS 5 5 26 Menu gt Export Positions Menu gt Import Positions
169. lIgnore 5 2 Excel Excel Excel txt do Biacore T100 52 5 5 Kinetics AFfinity System Preparatic ns Prime before run Normalize detector Temperature settings nalysis temperature 25 Sample compartment termperature 25 IE Help lt Back Next gt Llose Prime Normalize Temperdture settings Anqlysis temperature 29 Sample compartment temperature 25 C Next gt Freire HL Lat Fami a m 00 eiie 0 EF COC 4 OOOO CCC CE BT 1 1 1 1 1 CN MM MN YU CCOCOCOCT CC COCCOOOOO HH Epest Fisck Td Heas ad ul
170. list is nasitinned first Rack Vial Size gt Sample Smal Content gt Sample gt Small gt Aauto content gt Sample gt Smal Content gt Reagent Medium NO _ ctent gt Reagent Large gt Auto lt Content Pooling9 Yes OK Automatic Positioning Biacore T100 76 5 Eject Rack Rack tray port Hack Trau Ejected Llick OK to return the rack tray to the sample compartment Hep Time to auto close 58 OK Eject Rack Tray Rack Positions Start mmm ma aa ee se aw mm Start 9
171. nmimarnds Rapart Paints Carry aver control Comimands Repart Points a mple Ernhahncement Rieegeneration Ciarry ower control Insert Ciomrmahnd Ramanwe Frnm ylg Mnwe Lp Mnwe Lam 5nlyent correction jmject nHREcnwer eneral lt then neg Do 3d Cinntiegure settinegs tor each coitahnd Insert Command Biacore T100 6 159 Move Up Move Down 9 Capture Saumple Regeneration Sample Evaluation_software Sample Types Low sample consumption 7mm 28 ul High performance
172. programs Biqcore T100 Control Software KAT NT ES 1 4 3 Biacore T100 Control Software 041110 Exercise3 blr Fle Ed View Run Tools Help EE Cucle 1 Curve Sensor gram Fc Menu bar gt 8 Toolbar Event log Sensorgram Event log window Report point Keyword table table Status bar lonine CQ1 Temperature DD nC Sensor chip C5 sample Lompartrment temperatures curtent CU EE gB IRunning stanHdhy remaining Fime 3 4 daws Menu bar Biacore T100 Toolbar Sensorgram window 8 Report point table Event log Stqtus bar Temperature Sample compartment temperature Run Biqcore T100
173. re 4E lt Valule WWalule WWalule Walue 4 Tools PFPLLFLLPLLPPLFPLLPLLPLLPLLI PLLLLJ nlnr hw k Value irwes Fittineg Fk Ranking 4 Rannking RU RU One Ranking Boundary 1 First value Two Ranking Boundaries 2 First value Second value Biacore T100 5 95 2 RU
174. the list Cycle type description Cycle Types Sample amp Assay Setup Cycle types currently in method Concentration New Delele Copy Fename b 2 Insert the required commands fdr each command 3 Configure s tti 2 Commands Report Points Insert Command Remowe From Cycle kiowe Up Mawe Dawn 2 Settings for Sample 1 Ll Tupe ls variable Contact time on sl Dissociation time 5 sl 5 mm lon Solution Flow rate Flow path Piedip Mis wilh Fraclian of mis solution fh BIHilizafior plipd feT ig h ls Exhra wash after iniection with Slabilization period Cycle types 4 Melhod Variables Evaluation Variables Set property as yarlahle Salulian Cantact time s Dissociation time s Ol Flow rate min 6 3 d Gohntigure settings Tnr each cnmimarid
175. 00 gt 30 OK 0 lt 5 OK Fc2 100 e30 OK 0 cms OK Fc3 100 30 35 OK 0 5 OK Fc 100 90 OK 0 lt 5 OK Leaks Fc1 3 lt 100 RU OK Fc2 16 lt 100 RU OK Fc3 10 100 KU OK Fc4 2 lt 100 RU OK OK BAD BAD Biacore T100 186 8 8 4 4 8 1 Standby flow A 65 ml 24 4 Status bar 8 2 Toolbar Eject Menu bar
176. 114 5 Thermodynamics Results Create x Calculated Thermodynamic Parameters MM yant Haff Fitting function want Non limear i Fitting function Eyring Linear Parameter 393 mm a amp H kJ mall 19 8 AKkmoh TAS km uo 19 0 A mal X67 5 3 2e 3 3 25e 3 3 3e 3 3 35e 3 3 4e 3 3 45e 3 3 5e 3 3 55e 3 1T 1K Parameters TT OT Chi 9 508E 4 EI RE BA I 1 1fMsK Eyring association 1 1YsK Eyring dissociation 8 8 10 86 10 5 4 11 8 2 11 5 E asa 12 E78 5B 12 5 7 4 13 2 13 5 3 2e 3 3 25e 3 3 3e 3 3 35e 3 3 4e 3 3 45e 3 3 5e 3 3 55e 3 3 2e 3 3 25e 3 3 3e 3 3 35e 3 3 4e 3 3 45e 3 3 5e 3 3 55e 3 1T 1 1T 1K Parameters Parameters Slope 3930 1 AMS Help Intercept 21 26 1 thMsk Re 0 9953 Slope 9469 1 s Intercept 20 24 ZSK Re2 D9980 lt Back Finish Cancel Calculated Thermadynamic Parameters Fitting function twan t Non linear Fitting function Eyring Linear En Walue Parameter Name kJ mall 68SRID A fass kfmol 3 A Fs ED 0 TAS ass kJ mall 3 AG aes kJZmo on Finish
177. 44 11 0 231 1 637 3 568 Ma 4 892 i od EE 4 33 19 0 06103 0 0994 2205 9188 231 0 03734 4 2 269 3 496 3 471 20 25 28 0 002814 Cycle number 4 dno MV adusted resporrse bincding 5 3 Biacore T100 136 5 5 53 Evqluation Explorer Plot Baseline Sample baseline 4 RU 10 Carry Over co_binding co_baseline
178. 88G 0 3776 0 09440 OO OO OO OO OO io i ivi vi oi oo iR ER ER ER ER ERIR OO OO iiO Oi OMWGO In nnn Unnn Scioioiwi rio iN Oo Sh wm mw Tr a Co Control Samples lt Back Next gt Llase Sample id MW Da Concentration nM ug ml Next gt 5 1 5 0 0 1 2 Samples Next gt Bample serie Beta2u qlabulin The sample serie should cantain at least one nan zera concentration that is to be run at least two 2 mes lgnore
179. BHH 400 200 0 200 4dOO BIH BU 1000 1200 Response Refh RL Biacore T100 5 133 5 51 Solvent correction Include i Included Cycle Curve Chiz RUz2 YU RU 30iFc 4 3 0 4273 5 52 DMSO DMSO DMSO Solvent correction Extrapolate
180. C rein Cor tt be Re ina em i Regeneration command Pa Menu bar Commands Regeneration Regeneration EE xX wialygll pasitiorn m Corntact time ln Carcel High wiscosity snlulinm Help imirmum required nlume In wialewell For this imgchnn 37 ul 30 s 60s Ok Cqancel Biacore T100 44 4 New Cycle 2 me
181. E Sa re oe eee ss rwrm ee Save in File name Saqve ee Standby flow Biacore T100 Evaluation Software e Biacore T100 5 91 5 28 Run gt Stop Run Biarnre Ti x 2 This will stop the run Help stop Run ar stop Run Fun Stopped FinishIn chrrernt cucle please Wal bort cnclg bn Lt Bre Ctrl Break Biacore T100 Evaluation Software Biacore T100 92 5 5 3 2
182. Export gt Result To Excel Save in Bia Users lt e data 3 Methods and Templates History nakano da 9 4 YOSHITANI Desktop Mu Documents ky Computer lt a File name gt Mu Network F Save as type Micrasoft Escel xls Cancl Evaluation Explorer xls Excel xls nMea Mooertas fooeine faci Wmdme bes Bnd stskainy Beding te reterence aiaia css ur Biacore T100 9 199 9 6 File gt Save As Carve Results From kun As wE IT Bia lsers data ethnds and Templates Tiak nn HI Th My ncuments Mn Lnmhuler Mu Nehwark P Fe name awE as HPE Biacnre TH Eyaluatinn Files tbme Lance rw Save in C Bia Users File name Save 9 2
183. GE Healthcare BIQCOre T100 GE imagination at work 1 3053 32 AL A A CE 1 1 1 kk 1 PE OD Eki hp 1 1 1 2 ha 2 1 1 3 0 EEOR K KRV7NKX 5QQONRNRNQQN AAOJObh 4 TAT DM RR i i 5 2 VEN cl fl 5 CET Cf 9 hm TS ROCK 10 C24 AOR NG jl a 11 2 ns 14 3 jn 15 NO A 17 2 N EAN 2 8E K 19 5k iA ll 20 NIA OR 20 2 3 20 ME 21 A RE TIVITIES Jl 23 3 1 1 pH i 25 952 EC DE jj 30 23a EE oo 36 4 uaauiiiiiiateatessesteseestestesteseeeeeeteeeeeeeeeeeee 39 Biacore T100 5 kk 46 5 1 kk 47 49 5 1 2 2g 56 5 1 3 ee 65 OE EEEPC 69 OT 0 Ni Gd i i 70 JIJ 78 5 3 ee 85 e 2 6
184. Keyword Table Finish EE aywnrd Tablm CYS Bety Shep Pap TIE Pa CmmTeTDTi Biacore T100 5 79 5 19 Menubar SNEO rgram rdg mtshilrfr Summs 1 113TmO Bdne ts referemee Sames 1 Evaluation explorer Work areq Menubor Toolbor Evqluation Explorer ensoro ram i Bqseline AbsResp Binding level Bqseline RelResp Binding stqbility Baseline RelResp Binding to reference Binnding level Work area Evqluation Explorer Biacore T100 80 5 Toolbar Cohcentratiah hale a Concenwation Analysis Create Calibration Aiontral Samples Samples Calibration curye se
185. M5 3 BR 1006 68 Series S Sensor Chip CM5 Certified 3 BR 1005 30 Series S Sensor Chip CM4 Certified 3 BR 1005 34 Series S Sensor Chip CM3 Certified 3 BR 1005 36 Series S Sensor Chip C1 Certified 3 BR 1005 35 DNA Series S Sensor Chip SA Certified 3 BR 1005 31 Series S Sensor Chip HPA 3 BR 1005 33 Series S Sensor Chip L1 Certified 3 BR 1005 38 His taqg Series S Sensor Chip NTA BR 1005 32 Biacore T100 1 9 1 2 2 Menu bar Tools gt Prime Tnnls Help Prime Shutdown Biacnrg T1HI Ewaluation Start pn Flace butfer nn the left hand ran and insert tube a Flace water nn the right hand Iran rd insert the water inlet tube Prime Friminn please wal Time lett HL HE 3 The Frime hrncedure is completed Close Standby flow 1 8
186. OK Eject Rack Tray Rack Positions Start ere tes HH hee Fat EE LE mw mw oe Start Save as Methods qnd Templates Bia Users Dont Save Save in File name Saqve de Standby flow Biacore T100 Evaluation Software Biacore T100 5 _ 107 5 36 Run gt Stop Run Biarnre Ti x 2 This will stop the run Help stop Run ar stop Run Fun Stopped FinishIn chrr
187. Tools Normalize Start 9 OPT This nracedure narmalizes the detectar siqnal Total run me is about 9 mirnutes Hequired solution from kainternance Kit Binormalizing solution Next gt U Start Nomalize CC Narmalizina please wait Time left II UG 17 hmlce j PP The Narmalize nrncedure is completed Standby flow Biacore T100 184 7 7 3 Desorb and Sanitize 1 Biacore Maintenance Kit type 2 BlAtest solution HBS N buffer 100 ml 10X buffer Series S Sensor chip CM5 1 5 m
188. Tray Rack Positions Start Biacore T100 6 169 6 6 Control sample R1A1 R1A12 12 Ee Menu Automatic Positioning S 4 NT Ti I EO I TH Tee WE O 4 Se erg pli i re Pharthend A EE Fr lrwmml Chm em ote Fi Frh nm Es mE Fr rd rate Th Fiirmmam Them Eel mm Hail ff Cotoe Frierntas Miam 3 mr i TT ck WRm ml Cr c EE tn lr rim Er ET ri Er i Fmt AG rm Fint Frercr ng mmm t rie 3 Auto First reginn in the list is nasitinned fir
189. al RU NNUNUUNNN UE Il x Chip CM5 Response Response Flow cell Procedure Method Liqand Bound RU Final RU 2 Time and Flow Amine ani Beta2u lahlin 11141 4 8957 7 Help Frirt _ gss 3 4 Response Bound Final 2 Bound Final NHS EDC Final Bound NHS Finol Bound Oe Biacore T100 S6 8 3 1 3
190. e oc cp Assay Setup 2 Insert the required commands 3 Configure settings for each command Rename Solvent correction 0 1 Create a new cycle type or select an existing from the list Cycle types currently in method New Thermodynamics Delet Snlvent correction _ Delete Copy Rename 2 jmsert the required commands 3 Configure settings for each command Commands Report Points Salyent correction Carry Over control Snlyehnt correction Maye Down Commands Solvent correction Insert Command Solvent correction Assay steps New Biacore T100 6 Copy St Smulate Cycle Rn List 1 mw wberad Control amDle 2 Control are Thenmodranes Stertup 3 Statuo Thermodames 5 tmeas as ritered EE Thermo 3 Sarcie
191. ehntered Thermn 1 5arnhlg Thgrrnndyharrnics 1 time as ehntered Control sample 1 Control sarmple Thgrrnndyharnics 1 tirme as entered Every 15 cycles EJ Thermo 1 Every Distribute Run ossay step once first EE Run assay step once first 1 Biacore T100 156 6 RBrurrEhCE Ilw Repeat assay step withn Therma Eyery 15 rr CyElg Listrihute ncCUTTETHCES ByEHly Fun assay step once first Fun assay step once last Simulate Cycle Run List 6 2
192. ent Hep Time to auto close 58 OK Eject Rack Tray Rack Positions Start ee emmaam gl um ha an Start Save as Methods qnd Templates Bia Users Dont Save FM ramwr kcorcerwaimtm Bs 8 FE Imusr Cr cen Save in File name Save Standby flow Biqcore T100 Evqluation Software Ee Biacore T100 5 55 5 5 Run gt Stop Run Biarnre Ti x
193. erlay gt Sensorqram zoom lock 5 13 Keyword table Tools gt Keyword Table Finish Reset All Filters buffer hdd Keyword Warfarin Rename Keyword Warfarin Wartarin 0 308 _Femeve Keyword Wartarin 5 Wartarin 125 Warfarin 6 25 Warfarin 3125 Warfarin 15625 Warfarin 72125 Concentration Unit Vartarin 35 Naproxen 0D Naproxen 100 i Naproxen 5 Naproxen 25 Naproxen 8 0 Naproxen 625 i Naproxen 3125 Naproxen 15625 i Napraxen 78125 Sample Naproxen 125 Salyent correction Sampnle Furosemide Sample Furosemide 20 10 Finish Cancel Biacore T100 66 5 Toolbar ebest Blanks from other sample 9 hata 2 mikra 1 Sample Show concentration series Show blankS Show average blankSs ep we sample
194. ernt cucle please Wal bort cnclg bn Lt Bre Ctrl Break Biacore T100 Evaluation Software Biacore T100 108 5 5 4 2 Evoluation Biacore TIO0 Evaluation Software C Bia Users Templates demodata Thermodynamics blr 135 TREE Evaluation Explorer All sensorgrams Hi x 8 lt lcurve Name Fc 4 3 3 lt Assay Step Furpose lt Overlay gt F 24 Cycle lt Overlay gt RU Sensorgram I Zoom Lock 2 Sensargram Plot Baseline Sample a Binding level Binding stability ig Binding to reference Thermo Steh 4 Thermo Steh 5 Startup 5 Startup 1 Thermo Step 3 Thermo 1 Thermo Step 2 Startup 2 Startup 3 Startup 4
195. etics Affinity New Methods and Templates Open Browse Open Caplure SAMIPLE Sample REGENERATION 1 Heqeneralian lt 1 Capture 2 4 Saqmple Regeneration 1 2 Next gt CN Biacore T100 50 5 FE kinetics AFfinity Setup 3 Flow nalh 5tartun Lighgchinh Bun shartun cucles Solutior Bui
196. fer umber of cucles 3 Detection Flow path 2 1 4 3 Startup Solution 3 Next gt Kinetics Arfinity InjeCHOn Raa x a Contact tme 120 f s Flow rate 30 mmnl Dissociation tme 300 Er go Contact tme 30 ls Flow rate 30 plamin Stabilieation period ID _ Hep _ lt Back Ne pee Sample Contact time 120 s Flow rate 30 ul min Dissociation time 120 s Regenerdtion Solution High viscosity solution Contact time Flow rate Stabilization period Next gt Biacore T100 40 30 s 30 ul min 0s 5 51 0 000 0 02360 0 04720 Dn 09440 0 1
197. g 1Immobilization pH Scouting pH pH 5 200 ug ml 10 mM Immobilization pH Scouting 50 mM NaOH Biacore T100 26 8 Toolbar Run Wizard lt Menu bar Run gt Wizard Surface Preparation gt Immobilization pH Scouting New
198. globulin Temperature ho Curve Fe 4 3 2 Themo 1 i i 1 lB2ugobuin On Themal 3 120 32300 12 B2 u glahulin i Therma 1 IF Show cancentration series IF Shaw blankS Show ayerage blanks Multiple Rmax ON Bac Next gt Cancel Temperature Temperature H Conc hn k k Toolbar Thermedynamies Import Thermodynamics Biacore 1100 5 113 EL TE OE TE 6We7E E 1457E 9 3109E w5OTEe5 EE 0OWE 1 GAEDE OED En TE ED Next gt Thermodynamics Overyiew Create EIC esseeeeeeeeeeeeeeeeeeeesaeaeeeeseeeeseeeeeeeeeeeeeeeeeeeeeseeeeeeeee esseeeseeeoeoeoeeeoeeeeeeeeeeeeeeeeeeeeetcoeeeeeoeoeeeeeeeeseeeeeeeeeeeoseeeeeooeoeeepoeeneeenoeol 25 Temperature 15 20 25 Temperature Next gt Biacore T100
199. high2 Ctrl re re Hare Fe d 9 Reser Glep Papsss Lesined Oe eke eet RU SENSOTArAm Biacore T100 9 193 9 3 2 Sensorgram window 5 een Tools gt Color By gt Sample i Or Seem Lock Tools gt Report Point gt Id and Marker 9 3 3 Sensorgram window 5 DV Tools gt Sensorgram Adjustment
200. iacore T100 3 31 NHS 10 ul min 7 Aim for immobilized level Specify contact time and flow rate Blank immobilization NHS Specify contact time and flow rate 420 s 10 ul min Next gt 3 2 Specify contact time qnd flow rate 7 Custom Methods Amine
201. ion Contact time s Dissociation time s OO Flow rate plemin alue type Conc Numeric Dilution Numeric ser defined variables Name Value type elete dd Ewaluation purpose Concentration be Method Variables Solution Evaqluation Variables Evaluation purpose Evgluation software User defined variables Add Evaluation purp
202. l 40 ul min 30 Evaluation Software 2 II Solvent correction 30 ul min 30 5 5 Biacore T100 6 161 InjectAndRecover 5 6 If Then Method Variables Evaluation Variables 2 1 Method Variables Eyalualion Variahles Melhod Variables i Ewaluation Wariables Set property as variable Predefined wariables Solut
203. lewel Binding stability Binding to reference a Carry over ga Controls stability Report Point Table mm Report Point Table Solvent Correction A Solvent Correction running buffer Furosemicde Relative response binding 25 Cycle nurmber Evaluation Explorer Plot Binding to reference Biacore TI00 Evaluation Software C Bia Users Templates demodata Kinetics Affinity small maleculesblr Binding to reference rl Plet Evaluation Explarer lt lcurve Name Fc 1 5 lt Assay Sten Purpose Overlar lt lcycle number Overlay gt x Binding to reference 1 zaam Lock Sensargram nd sensargrams Plot gq Baseline Sample gag Binding level Binding stability inding to reference og Carry over ga Controls binding ig Contrals stability Report Point Table mm Report Point Table Solvent Correction Solvent Correction Startup Sample Control sample amp 25 Cycle number Biacore T100 98 5 5 32 Tools gt Custom Report Point ata REphesrl Pin Add
204. ment Regeneration gt REGENERATION 1 Help a Next gt Close Oi Capture EV NC EE 2 4 Sample 5 24 Regeneration 1 2 Biacore T100 86 5 Enhancement 2 Next gt x Detection Flaw path 14 3 0 Startup 7 Run startup cycles Solutior Bufer Number of cycles gt Help lt Hack Megl gt Close Detection Flow path 2 1 4 3 7 StartuDp Solution
205. ne pH 1 Bu 1 E40 47150 n 3 6 3 12 15 18 21 24 Cycle _ Hep Display Sensorgram 1steycle Candiions EE g sensorgram Fc 4 lt Help Print Close Application Wizard Assay Development Surface Performance 400 Eo a 5urface Performance Injectiom Paraneters 5 WDM nluiorr lmeusmlgi Contact tme ED Flow rate 30 mmml Binding stability _ I eg Curve Name Fc 2 1 ll Assay Step Purpose lt Uvellay gt lt Cycle rumber lt Uvellay gt Gah mM Han Binding stability X Value amp 4 Y Value 5nluhiomr rm Contact time 5 Flow rate 50 mm Help Ba Biacore T100 46 5 5 6
206. nish Add Fit Mode Fit Current Fits mn State Reaction kontor matiornal cha Current Fits Finish i 1 Rmax RI Add Fit Parameters Biacore T100 64 5 Rmoax Rmax Fit Fit local 0
207. orrection Plot type Report Point vs Variable Axis setting Y Report Point binding Response type MW Qdjustedresponse X Variable Cycle number TET Plat Settings Plot name hw correc tian Flat type Heport Faint ws Variable CC Repart Paint ws Report Point Axis setting Y Axis KAxis Report Paint binding Wariahle Cycle humber lt Response Type MW adjusted response Help Finish Cancel Finish Biacore T100 5 135 Eyaluatinn Explorer Edit Rg mowe GETSnFETaI a nd All sensoregrams Flot Baseline Sample a i Binding lewel i Binding stability i Binding to reference 0 FF Carty Over SS Controls binding i Cantrols stahility Be Flnt Soleent Correction 0 1 Solyent Correction Evaluation Explorer Plot Plot Work areo Poet 44ICurye Name Fc 2 1 corr lt Assay Step Furnase Uyerar 44 Cycle numher Uwerlay gt gt 4 100 RLUDa Zoom Lock 4 0 2637 0 2081 9 0 08493 0 61
208. ose sqmple Evaqluation purpose 6 Kinetics Affinity Concentration Affinity in solution Thermodynamics Kinetics heterogeneous qnalyte General Biacore T100 162 6 Report Points Contegure settinegs tor each Com mahnd Before After Start of End of Inject Window Baseline Biacore T100 1 Betore Start of Cabture 1 5 YES Er End of ahtUF 1 1 n Betore Start of Samble 1 5 ES 5 Beatore End of Gamble 1 1 BT EF dn f 5arnIB 1 Start of End of Inject Start of End of Inject
209. s referemee Samem 1 Evaluation explorer Work areq Menubor Toolbor Evqluation Explorer ensoro ram i Bqseline AbsResp Binding level Bqseline RelResp Binding stqbility Baseline RelResp Binding to reference Binnding level Work area Evqluation Explorer Biacore T100 94 5 Binding level Evaluation Explorer Plot Binding level Lee Name
210. st Rack Vial Size gt Sample Smal Content gt Sample gt Small gt Auto content gt Sample gt Smal Content gt Reagent Medium NO _ ctent gt Reagent Large gt Auto lt Content Pooling9 Yes OK Automatic Positioning Biacore T100 170 6 Start Save as Methods and Templates Bia Users Dont Save Save in File name Saqve
211. ttings Flow Fc lt Report point stability lt Respanse type Relative respons Fitting functior 4 Parameter lt Calibration table Calibration curye Conc Respons curve 1 ef e RU L em D RU mouselgG 5 3500 Soo Parameters for curve 1 Rhi 2693E 04 Rlo 1507 Al 4542E 04 A2 2D00 D 2000 4000 GOOD SOOD 10000 12000 1 nan Concentration nd Help Finish Cancel L Concentration Analysis Create Coalibration Calibration Calibration curve settings Flow cell Report point Response type Slop Calibration curve Cryg Lalihratinn Curve lt Ctrl
212. ve qs Methods and Templates Bia Users Dont Save Save in File name Saqve Standby flow 1 s 3 3 Ctrl Break l Biacore T100 Hi me TI Lnmhrul llmramre ImimighiiraLinm mrmie 1 Ee Ek Yr Hum Teo bol Terns 4 ariti WWta lnuBlin NHS EDC Ligand 1 Ethanolamine Fc Tamal Woee AbuRep SD LRSD Same Refempl aes 5 m0 3 0OG 2 OU Ye Hi CCa 1 a mmo I se g Ha EE HE i EH 3 3 Ep 5 PrPrclii EF EF TE EE 11 1 mW Lr 1 1D 5 Hg rmmnLmre Ligmrmd Pa Ta Tm re Em rna COL Temperahurai 5 00 WC Sareer hg CMS TE si Eri hh Response Fin
213. w rate plemin Stabilizatinn period ln s Help lt Back Next gt Close Sample Contact time 300 s Flow rate 5 ul min Mix with Biacore T100 72 5 Regeneration Solution High viscosity solution 40 2205 Contact time 60 s Flow rate 30 ul min Stabilization perion 0 s Next gt Te Concentration Analysis Calibratioml NE x Calibration urye Analyte name Imousel ai Repeat calibration every sample cucle Calibration points Concentration Help lt Back Nexl gt Close Calibration Curve Analyte name Repeat calibration every Calibration points Concentration 6 2 Next gt
214. wgll position TR1B1 Contact kime lg Laicl Help imimum renulred nlumg In wlal wgl nr this imectinhi 8 hbl Vial well position Contact time Injgect fm acore 100 Contrel Softmare te sa gE x x ga Eg Cewe 1 De Sentongsn lt 1 IT a Lock seale 35270 met Vi posse cr Lorded mm aa Mewmwun peOJmed Le oreTeY 7 XY CT OT 35540 t CO Te o 19 20 30 40 70 0 Fc Tme Wngdor AbcHop SO LRSO Skpe Aofioip Eseine Kepts n cre 1 VA ol OOC Te Paw 20 Flow pate 1 lomne COML Tempershure 25 00 0C rm marnsal um SCn Cowowrtnmmwt mraure Cm 5 si ST wacees To cewrol s Ma0 Peet PLALA cm Caqncel Inject Eject rack tray Menu bar Commands Eject Rack Biacore T100
215. word table Tools Keyword Table Finish Feset ll Fers 15 So herent cmmreetim 1 nm 1 Samp 1 mm 19 Sampke amp Sumpke Fr ET i Samake EL ET rl wT am ample Rn mple EE Biacore T100 130 5 5 49 Menubor Ve ce ES Toolbar Evaluation explorer Work area Menubar Toolbar Evaluation Explorer ogram jpiot Baseline Binding level Binding stability Binding to reference Work area Biacore T100 EO ram Sensorgram window AbsResp Baseline RelResp Baseline RelResp Binnding level Evqluation Explorer 5

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