Biacore S51 日本語取扱説明書
Contents
1. Biacore S51 Instrument Handbook 50 3 Next gt hl Hit Sglectinn hssay oetup ea ear rmwtagh2. CN hm Pe
3. OFF TEL 03 5331 9336 ED 9 00 17 30 WWw geliftesciences co jp e mail
4. DMSO DMSO DMSO Biacore S51 Instrument Handbook 3 33 150 RU 1 Spot
5. 1 Biacore S51 Instrument Handbook 16 2 CM5 certified grade 1 EDC N etnyHN 3 dimethyloaminopropycaqrbodiimide hydrochroride NHS N hydroxysuccinimide 1 M ethanolomine hydrochloride pH 8 5 EDC NHS 10 ml MiiQ
6. Export Curves Text Exel Biacore S51 Instrument Handbook 3 89 3 3 Compound Characterization k k Kp Wizard 3 3 1 Menu bar Run Run Wizard Tool bor DD 0 Run Wizard Sutace Preparaton rrrmcibiizaticor La immaibiizaticn pH Scouting FD ss Dewekopmernt Iriearirwe Fi Bindinn Lnniharrs Hanermmialinn Lnndihnrs Fit Hun ssays Hi Perfornms birding sssays Inr anhalte corncertrahorn senies designed fo measurrg the kinetics rynr affini
7. sc 1 Biacore S51 Instrument Handbook H9 3 NN OU 6 DR 9 F OD
8. Binding levels RU Da menu bar Evaluation Report Point Corrections Repor roint Corectons kalscular weight correction Ta compensate for the fact that a large molecule gives a higher response than a small one the response lewel may be divided by the molecular weight ges Biacore S51 Instrument Handbook 66 3 Activate weight correction Next gt Report Foint Gorrections x dstmernt For controls Report Point cis ep Edit Delete Back Llansg Finish Evaluation Explorer RU
9. Menu bar Run Run Wizard Tool bor Run Wizard _ _ Sumface Preparation Immnbilgahnn Immnbllgahnn pH Scouhrg Mssay Dewelopmerw FF Interactwe Fun Ee bE FH Regqeneraton Corndtions Piot Riun Assars Hi Selmcinn Compournd haractelizalinn Tests birding of arabate to the surtace al two diifenerk corcertrahotes arsls cycle Adibonal crcles can be added wathin the same ru Use this izald to test sface actrity an get a first estimate of the aralyte BI characterics Start Wn Birding Conditions 5etup Detecticn Flow cal 1 Spatl1 Mae te Flow dk llymmnl Hare Furasermide Cantact tme lsn Dissociaton trmec 300 Concerirathor Il pectior 10 luM Lnncerrahn secord inechorc 5 bd Biacore S51 Instrument Handbook 3 37 3 Detection Flow cell Spot
10. 100RU DMSO DMSO 16 1200RU 10RU
11. Menu bar Tools Eject Chip Tool bar Eject Chip This wll eject the sensor chip Help Praperties Eject Chip 2 Insert Chip Insert Chip Chip huhe Chip status New chip Beused chip ew chip Chip id 50225 1 1 2 Chip Int nn optional Help hinck Chip amal Dock Chip Chip type Bidcore S51 Instrument Handbook 114 4 4 1 Tools More Tools Desorb Maintenancw Tnnls Desa EDesorb rd Sawce Sup
12. residual plots residual plots Y 0 Biqcore 2 1 1 Moss transport limitation MTL MTL Check Kinetic Data Hm I Hm Sh Se Krein summasl inalicg Fea Spoted r coorT Biacore S51 Instrument Handbook 102 3 Check Kinetics Datq Check Kin
13. Description Sensorgram Report Point Add Add hnundary negative cnnlral Respornse l IHU Help Lanrgl Name Response OK Ranking plot ORU negative control sh Add Ranking x Name vs negative positive control Descriptior Sensorgram Fc 1 Spot 1 r corr Hepart Pain binding_late Hankinn baundaries Control samples hedatiwe cnrittral ED View average values View all yalues _ Edk Controlsample conc wy awg Delete U D 4 Control Furosemicde SD BE Hankina plot 10 Cycle number Allaw in graph editing of boundary yalues Sort by cycle Number Help OK Cancel Biacore S51 Instrum
14. Compound Characterization Biacore S51 Instrument Handbook 3 35 3 1 3 1 1 Change All Solutions Stqndby Tools Change All Solutions Hacore 551 Lmntrnl 5 nftWarE mn CO RH Change All Soluthonms Place the soluitioeee in eagert comparmernt rc ireseit he la hubings Start a Change All Solutionms Te Charge All SohAions procednes 5 compieied E E Pi Ea 4 30 Close Standby flow Bidcore S51 Instrument Handbook 36 3 3 1 2 Binding Conditions 2
15. Biacore S51 Instrument Handbook 74 3 Binding to reference spot stability_early baseline RU Binding to reference spot 7 _ O 6 ao oc gt a O 5 SS gt no ge 8 9 4 9 9 O a gt Do O C gt gt CO 3 DTT o gt cS gE Es EE 9w LO lt oc GGE oS E LO o Go 9 SEGo 8EG TELs amp 8o DE JESc 85 5 Eo omLS EGF CI _ EEOS Bo 6 UE EE spowE Fc amp SE Fg DO SIES SG POSEE BD 6 o XAGO GOT Sm Ss Ps 6O o OCNGOGG OO 6 O G TO 1 CO gE cc gt STE OSC Oo SQ 8 _ cec 9 0 RN ES GEoES CO GOD GEGg se X No EE Tg EO SODS DBs 0gIOEITGD PMs ES 8 5E 1 LO A TN NE ET NN 2
16. RI RU RI Constant 0 Biacore S51 Instrument Handbook 3 105 Rmax Local Kinetics evaluation Advanced dranced Kinetics Settings 3 Fr 2 5hat 1 cor Fc 2 Spot 2 T Cor Both spots combined Kinehc Farameter Settings ritial walue Fn ffset durin inection Rl Lncal Inihial yalue For kIg 5 EEer Help Larncel Rmax Local Advance Kinetics Settings _ Fc 5hot 1 T cor Fc 2 Spot 2 r cor Eoth spots combirned Einetic Parameter Setthings ritial walue for ka a Imlal walue For Ed Offset durin imactinh RI Lacal ritial walue For kh IEEez Hep Caneel OK Biacore S51 Instrument Handbook 106 3
17. Dry 4 C Wet HBS N 25 ml 4 C Sensor Chip CM5
18. Remove Selection Next gt Biacore S51 Instrument Handbook 3 97 4 Add Compound Characterization gE x Compound characterization name R50 evaluation Fc 1 Spot 1 r corr Fc 5not 2T cnTi Stead state eyaluation lt Ine site madel tnesite mndgl Iv Fc 1 Spot 1 r carr Fc 1 Spot 2T ca Muliple site madel IE sitE ml Kinehics evaluation Fc 1 Spot 1 r corr Fc 5pot ZT Ca Bothi snots Carnbiried advanced Help Zoom Lock lt Back Finish Llase 99 Finish Compound Characterizatinns 8 Eompound Characterizations i iglete pliwalE Deactivate Add Close Evaluation Explorer Compound characterizotions
19. DMSO DMSO DMSO UV spectrometry DMSO 10 DMSO pH DMSO pH 150mM NaCI Hepes HSA HSA Hepes DMSO
20. DMSO 32 50 DMSO Biacore S51 Instrument Handbook 3 31 1 hot 1 hot r BBuffer 5 hot H Jl Eo JI usr O NE Biacore S51 Instrument Handbook 32 3
21. id Chip id 6 Dock Chip Run Wizard Standby flow Stqndby flow EE WW Sumface Preparation Immnblligahinf pH SEnum Time s HH IEB RU 0 Assay hewelnpmerw a UR Irieraclwy Hum Bindig FandWians FIa ce 1 ei oc Temperatun CC nt um Assays Sitandbhy How max remai HE Selection Compourd Charachenzalion Frernhcarns cowaleri mobilzakio of lnand nn He semsnr sufacs The sa nf farerW hgards can be immahilced on bch detection spots In the Fa cell im a sgle Fu he se Template _ sax a Run Wizard Start Biacore S51 Instrument Handbook 6 1
22. rr Immobilization pH Scouting 5 50 ug ml pH 10 mM pH 5 0 100 ul 10 mM pH 4 5 100 ul 10 mM pH 4 0 100 ul 50mM NaOH Immobilization pH Scouting Biacore S51 Instrument Handbook 2 19 Menu bar Run Run Wizard Tool bar
23. 1e 3 Fc 1 Spot 1 r Farmula Riesponse Conc Fimax1 A Conc KD1 Conc KD1 1 253e 5 M Hmax1 49 02 HU KD2 gt 5 000e 4 M Hmas2 Chi2 5 373 RU2 1 Kp1 Rmax1 2 Kp2 Rmoax2 Kp2 gt 5 000e 4 Kp1 Kp2 Biacore S51 Instrument Handbook 3 101 F Spot T Frrmula Respornses Conc R ma AConc k TCornce imane A Conctk Dz EDT 1 260e E hd Hmas1 7 355 RU 4 993e 5 hd Hax2 32 89 FRU ChF 1 345 RL Kp Rmax Rmax Rmax1 Rmax2 10 2 Rmax one site model Kinetics
24. RUUJ mol mol RUJ Rmax RUJ RU J Rmax 1 1 mol mol Biacore S51 Instrument Handbook 2 17 RL MWL Rmax MW MW Do MWL Do Ru RU Rmax RU Rmax Rmax RL MW MWL Rmax
25. Biacore S51 Instrument Handbook 34 3 Wizord Change All Solutions Binding Conditions DMSO Interactive Run Hit Selection Compound Characterization Change All Solutions Binding Conditions DMSO Interactive Run Hit Selection
26. 10 7 A706 RU Heb Hem Cotse twee Legend ridings nnrTi Lnck y ho Rennrt Fnint Ranggs Extrannlatinn Rangg GBTIS TET ITIE HGnlyent Correction Extrapolation Rarege Ewtrapolation Hanne w amlg RU Help Farncel Response Ref OkK Bidcore S51 Instrument Handbook 64 3 3 2 3 Evaluation Explorer Result plots RU Sensorgram Azosulfamide 50uM 3980 gt 2 gt G 3960 a 8 2 gt Cc 3940 _ MI oO o 3920 1 3900 G MP 2 _ QG 3880 Pe 8 PV QG 3860 0 50 100 150 200 250 300 350 400 Time File Edit View Evaluation Tools Window Help slSensoreram x Evaluation Explorer Sensorgram Buffer Sensarqram Q
27. PBS 10x BR 1006 72 100 mM phosphate buffer 27mM KCI 1 37M NaCl 27mM KCI pH7 4 HBS EP 10x BR 1006 69 100 mM HEPES 1 5 M NaCI 30 mM EDTA 0 5 Surfqctant P20 pH7 4 HBS P 10x BR 1006 71 100 mM HEPES 1 5 M NaCI 0 5 Surfactqnt P 20 pH7 4 HBS N 10x BR 1006 70 100 mM HEPES 0 15 M NaCl pH7 4 HBS N BR 1003 69 10 mM HEPES 0 15 M NaCl pH7 4 HBS EP BR 1001 88 10 mM HEPES 0 15 M NaCl 3mM EDTA 0 005 Surfactant P20 pH7 4 HBS P BR 1003 68 10 mM HEPES 0 15 M NaCI 0 005 Surfqctant P 20 pH7 4 0 22um Biacore S51 Instrument Handbook 1 3 1 1 2 Start BIA programs Biacore S51 Control Software Biacore S51 Control Software on COM1 2 Compound Characteraization_blr Sensareraml x i File Edit View Run Tools Window Help x 2 Cucle 1 Lurve blracted Fc 2 Spot 2 r 2 RU Toolbar 0 Jr 3IIH
28. 1 Select cycle no1 Startup Buifer Yes II2 Startup Buifer Yes II3 Startup Buifer Yes II4 Startup Buifer Yes II5 Startup Buifer Yes IE Salyent Correction Yes 07 Control 5 ample hegative 5 DMSD Yes Control Sample nenalive 5 DMSD Sample LBS amp Do Sample CBS amp Yes 1 Sample phenytoin Yes D12 Sample phenytoin Yes 13 Sample azosulfamide Yes Di4 Sample azosulfamide Yes D5 Sample warfarin Yes DE Sample warfarin Yes 17 Control Sample hegative 5 DMSOD Yes Ds Control Sample hegative 5 DMSD Yes EE Sample sulfanilamide Yes D20 Sample sulfanilamide Yes D21 Sample haproxen Yes 22 Sample haproxen Yes 23 Sample furosemide Yes 24 Sample furosemide Yes I25 Sample digitoxin Yes I25 Sample digitoxin Yes 27 Salyent Correction Yes I28 Control Sample hegative 5 DMS Yes 29 Control Sample hegative 5 DMSD Yes 3D Sample CBS amp Yes I31 Sample CBSA Yes 32 Sample phenytoin Yes 33 Sample phenytoin Yes 34 Sample azosulfamide Yes 35 Sample azosulfamide Yes I35 Sample warfarin Yes gt nF ranle TE ori Mn Hoo OK Biacore S51 Instrument Handbook 82 3 bana
29. Ko ka k Biacore S51 Instrument Handbook 68 3 Menu gt Evaluation Rankings Rank ings Hame lescription Curee IEeportPomnt Hd Ed Halsle Add 1 Add Rankine X Name vs negative positive control Descriptior Sensorgram Fc 1 Spot 1rcor gt HepotPonk 00S Fianking boundaries Control samples dd View average yalues View all yalues Em govrosample comewo lawg hE edative control EF Cortrol Furosemicde SD sd Hankina plot 8 10 Cycle number law in graph editing of boundary walues Sart by Cycle Number es Help Cancel Biacore S51 Instrument Handbook 3 69 Name
30. Biacore S51 Instrument Handbook 98 3 alazosulfamide 0 RU R50 Fc 1 Spot 1 r corr R50 VN UNA AI R50 Fc 1 Spot 1 r corr Farmula Fespanse Rhi Rhi Rlo 1 ConeA1 2 Fc 1 Spot 1 r carr Rhi 84 5 RU Rilo 2 B5B RU R50 0 4273 ph Chi2 0 09347 RU21 Residuals Fc 1 Spot 1 r corr Check Kinetic Data Biacore S51 Instrument Handbook Steady State pe R50 Steady Statel Kinetics Summary Steady State Fc 1 Spot 1 r corr 4e B Be B Be B 1e 5 1 2e 5 1 4e 5 1 5e 5 1 8e 5 2e 5 Conc Fc 1 Spot 1 r carr Formula Hespanse Conc Rimas Conc KD offset KD 4 085e 7 M Hmax 85 78 HU Dffset 0 4515 RU Chi2 0 7902 RU21 Summary Fc 1 Spot 1 r corr 0 4273 Mi II9347 RU2 Steady State Evaluation Fc 1 Spot 1 r corr HBBe 7 Mi 0 7902 RU2 Kinetics Evaluation Fc 1 Spot 1 r carr KD 2 gEHe 7 Mi kd 71Be 3 is ka 2 BH7e4 1hAs Chi2 2 252 RU21 Fc 1 Spal 1 r carr Check Kinetic Dala 3 99 R50 evqluation R50 Fc 1 Spot 1 r corr
31. Insert Chip SO hp Ehpbpe_ lrv5 _ Chp ui ew Ehip Rased chip Chip status Reused chip id Properties FT hip Properties Chap dl EI First ss al FU tppe TE FE1U Flnw Ligand Immnh Immab limmmnmh file Comment cell a te lewel amd_ 2717 2n03 3 jmnnnb hd bam as immob Biacore S51 Instrument Handbook 1 7 1 2 2 Change All Solutions Tools Pp All Solutions 4 30 Hiacore S55 Controal Softmare on CO Fis Edt Visw Run Tools Windnws Hep a E Eyalurainn Crl E J sa Fh Sn csster Charge All Sohutions Change All solutions Flace lhe soluliore in he ea Cnmihallmert TH hselt he In uF Start 4 30 Change Smkuthones The Change All Scions fee conpieled tune 25 00 C Mt Feairmirg hime 4 00 Days Biacore S51 Instrument Handbook 8 1 1 2 3
32. View average Next gt Bidcore S51 Instrument Handbook 96 3 hdd Cnmpound Characterizatinn Include sensnrnramm Fc 1 Spot 1 r corr Include sernsorgram Fc 1 Spot 2 r Cor EE electall Cucle 8 AzosLlfamide Op Cucle 39 Azosulfamida DLH7BM Deselect Al Cucle 4 zosulfamide BE Lucle 41 razosulfamide Tp Cucle 4 AasLlfamide EZ Cucle zasulfamide 1 25phd Cucle zasulfamide pi Lucle 45 zsulfarmide hphd Cucle E zosulfamide Tpkd Cucle 47 zosulfamide gn Hemowe Selection nn Fc 1 Spot 1 r corr eawe elechon Help nnm Lnck lt Hack Ma gt Llase Include sensorgram Include sensorgram 1
33. Gridlines ainr Gridlines nar Gridlines OK Legend Legend Fngsitinn Hidden Left Tnp Right BanttnrT he OK Bidcore S51 Instrument Handbook 88 3 CyEle 1 FC 1 Shnt 1 F corr Cycle 2 Fc 1 Shot 1 F corr Cyrle 21 Fc 1 Shnt 1 F corr Cycle 2 Fc 1 Shnt 1 F corr Cycle 23 Fc 1 Shot 1 F corr Cycle 4 Fc 1 Shot 1 F corr Cycle 25 Fc 1 Shot 1 F corr Crcle Eb Fc 1 Shot 1 F corr Copy Graph Paint Past Paint
34. Amine Coupling N CM EDC NHS NHS CONHCH CH OH Surface Thiol Coupling 4 0 PDEA S S PDEA pH
35. ImtErartiyE Rum Rack Fositions OOOOOOOO 68988898 Stqrt Biqcore S51 Instrument Handbook 3 43 Gr Ward Resulls is 5awe Ini binHiri condition am blnrhnn mialm ll Wi F A nH srnurW Desk tap in iocumeris i Carputet File riame hy Neltmork P av as type Resuks bl File name Suve Interactive Run wizard EL Belete Nw Lc ET HUT Hp Inject Biqcore S51 Instrument Handbook 44 3 HN Tteractive Run Injeet Solutiore nsgahys crirol lt Valums 3 bal Contact tme ED Iniect Solution Volume 30u 60 Biacore S51 Instrument Handbook 45 Plocrc 551 Cartrol Softmars en COML interactive ru Sensorgram x Fe Edit Vem Run Tons window He
36. SA Aim for immobilization level Next gt ImmTtOb zaHiin FITETTHTHE1 x Flow eal 1 snat 1 Ligand LA Inmmmobmlizahcwi method Amine Cantact ie 12 Imml Flow rate ME lt luMmml PCE hm Linan isahmm miafhni Nm Fwraglip hukmm Heln lt Back Hese Contact time Flow rate Contact time 7 15 Flow rate 2 5 10 u l min 3 10 umin Next gt Bidcore S51 Instrument Handbook 28 2 mrimolbiiatiom Sysherm Preparations Next gt EN Imobilization Rack Positions me Mieal BIAdesorb i 1 EG il BIAdasnrh solutinn 2 Bt 0 8 1 leMWe S32 Deionizedwater 0 II DI 2 1 Biacore S51 Instrument Handbook 2 29 Amine coupling Fc 1
37. Save slEit LhErk Rr 2 Time left raz ad 40 Biqcore S51 Instrument Handbook System Check System Check_ ame ate yr Biacore 51 Control nfhwarg Insirumenti 10129 Rn Dale rr Ahr D3 iE A i Readent ums 5 5 RU 4 OK 5m 1499 RM 3 1983 eOO3 OK 3 2080 1590 OK 4 2034 1997 OK Refractometer Bt eset eolutionn RI Fa1 St 1 ggd5g OR ef zz ldh pot ss 22083 OR Fc Spnt 1 ET 1 Rgf CT Fa shut 19847 Ha WEBTa 1BB St Flow cell leakage BE 2O0 OK CTINgE UMEs EN Coniigurafioe Eualuation rate T ET UE 4 121 1l 1 FC1 2 TA NDS la acelimne lml EODEE FL Ft 1 WP FE Ei Fl a EN Fa Fr EF 1 Ba OK Bad Bad Bidcore S51 Instrument Handbook 122 5 5
38. Flow Cell 2 Spot 2 Spot1 Spot2 Spot Flow Cell 1 Spot 1 SDOtT SDOt 2 Flow Cell 2 ar Biacore S51 Instrument Handbook 2 21 ILILILuUIIFGIIIUIIUIETm1 Hi Setup Flnwu and detection Eeemh sh Ligand hngs Name bormc hhydrase Conkact time 2 min FInWi al hn Mn mmi uaces egerearahor This regereratorn vil be peafoamed omce the er of the veand 5 lulgr IEnmM H _ Hep _ Pose Ligand settings Name Contact time 1 2min Flow rate 10 umin 2 5 umin Surfqce regeneration Solution 50mM NaOH Next gt
39. Add Edit R50 Steady State 1 1 Exclude Cycle RU R50 Fc 2 Spot 1 r corr 5 Exclude Gyclg rzoom ca Cnny Granh Exnnrt urwBs 15 ridlines Biacore S51 Instrument Handbook 104 3 Rmax RU Steady State Fc 2 Spot 1 r corr lnclude sgnsnrnrarn Fc g 5nnl T En 2 Lnclg 1H arhnspbenzehe sulhnnamide 1 5E2E
40. Ignore Biacore S51 Instrument Handbook 14 1 1 2 6 Standby Standby flow Menu bar Tools Stop Standby File Edit Wieww Run Tools Window Help a FN BIAsimulstion SE hilutiom ElmEnF oo C har ll 5nluHions hutdnwn qr 5 andhw Shop 5Fandhy Eject Rark BL E1e ChI 5 TEETFUW egBULU PreFETETICBS Flore Tools brl T FE 10 Status window Standby flow Biacore S51 Instrument Handbook Flow cell 4 Temperature 500 C 2 15 2 90
41. Web 2009 GE lt GE 169 0073 3 25 1 ISO 9001 2000 TEL 03 5331 9336 FAX 03 5331 9370 e mail Tech JP ge com
42. Standby flow 3 4 Biacore S51 Instrument Handbook 118 4 Superclean 0 22 um 196 acetic qcid 0 2M sodium bicarbonate NaHCO3 6M guanidine HCI 10 mM HCI Superclean Start superclean x This proacedure clears the Ho susterm Total ru hime is ahnut 0 irites HOTE lse the hainternance Chip For this procedure The surtace nn other ensnr chips may be damaged by the snlutinrms used se itered deiornzed warm water AOC as runnin buffer Next gt Superclean x Fiequired solutions 1 aretic aci sndium hicarhnnate laHLHI EM uanidine HLI 10 mh HHI Hack Next gt 16 mm i Ra 6 Superclean Rack Positions positon vo 00 _ Conem 90e OO ER Heaqent Hack 2 175D 02 M Sr bicarhnnate IR2A3 125 IBM guanidine HCI 10 mM HCI 81 Buffer IS2
43. Kp 1 Kp Afr Brree AB lt 1 Kp M Aree _ M Brree M AB M 1 Ai l Be Ko Atot AB Btot AB AB 2 Atot A M Bud B M Kp Atc Brosd 2 AB A bound 3 96 bound AB Atot x100 3 Biacore S51 Instrument Handbook 108 3 3 4 2 9 bound Compound Characterization Ko Menu bar Evaluation bound Tables sn_hmund Tahles Add nd hnund Calculation 3 Compourt ldicklenac Lnmpnund cancerilahiri In un
44. kJ Kk Kp ka kg Rmax Bidcore S51 Instrument Handbook 100 3 3 3 3 R50 RU RU Steady State one site model 1 1 ko R50 Rmax multiple site model 1
45. FInw suslam nieP ialinh Marne ei al a nl a Nl isn il Nomaize detector ralysis bemperatuae I _Bep eee Flow system preparation Refresh flow system Change all solution None Analysis temperature 25C Next gt Bidcore S51 Instrument Handbook 22 2 0 Immmobilization pH Scouting Rack Postions Rp Carbonic sm in 10 mM EE nH 5 i RB Se drase in late pH SU IR2B4 CKO CC OO OO Help Me Eject Rack lt ai Slant Close ul Vol up
46. HH 60 4 Z 2 a O O
47. 4 parameter 1 1 R50 B50 Steady state evaqluation Steady State Fc 1 Spo 1 corr Kp One site model 1 1 2e B 4e B Be B Ge B 1e 5 1 2e 5 1 4e 5 1 5e 5 1 8e 5 2e 5 9 Fc 1 Spot 1 r carr M lt ple site model Farmula Hesponse Conc Rimas Conc KD offset 2 Kinetics evaluation
48. Y axis Zero qt report point baseline OK Biacore S51 Instrument Handbook 86 3 mlSensnrgram Cucle coverlayy kk Gehsogram FC Sopot 2 r En Zaom Locki RU Adjusted sensorgram Gl 5 NN 8 nH 8 9 E se aD a ns IS es I Ei a 2 1 nh 1 gn 3HI 4 SHI EO TO Time th start mmlSensorgram Lucle snwerlaur kk GE T Ta Fc 1 nl 1 T corr nnm Lnck RU Sensorgram HS le nnw rahh Exnnrt Curwes 5 4nn THHines Leegend 5 3 Tims Scale Biacore S51 Instrument Handbook 3 87 T Gcale euto Lagarithmic W 0 E 0 Lungarithmic h on Cancel Scale Y Scale Auto Min Max OK
49. RI Constant 0 Kinetics evaluation Advanced dvahced Kinetics Settinegs ilnhal Replicate measurermehnts Cnlncidg Offset during injection RI Local dvahnced Kinetics Settings ilnhl Fieplicate measuremernts Coiecide Eonstint 0 I 5 bber OK Biacore S51 Instrument Handbook 3 107 3 4 Biqacore HSA co AGP Kp Kp HSA 30 55 mg ml 450 850_ uM AGP 1mg ml 24 nM 3 4 1 bound A B
50. Stoatus window Bidcore S51 Instrument Handbook 4 1 1 2 1 2 1 Insert Chip Biacore S51 E Biqcore S51 Instrument Handbook 1 5 Insert Chip EO chp Ehip re Chep status Fewe chip Heused chip ww hip Chip jd 030328 141 Chip lat no igplinnal Help Carncel Insert Chip Chip type Chip _status Chip id Chip lot noloptional Chip id
51. Rack Positions Start Biacnre 551 Biqcore S51 Instrument Handbook 2 File name Sqve Wizard Dont Save YE WW dFr HELLE s Desktop Docimernts PT 1 Mn Computer ave as pe Fesults FF bk Mu Na P N Save in C NBia Users File name Sqve pH Bidcore S51 Instrument Handbook Wizoard Save Save As 24 2 Biacore 551 Control Software on COMI pH scouting Sensorgram a x bj Fie Edit View Run Tools Window Help g Pp mw mwl sssesrosen PH scauting Carbonic anhydrase Fc Spol APROG Time AbsHesp Slope Baseline HelResp ld Flow Vol Name 1 1 pHscouhng 200 291605 004 Yes 0 baselinel 10 20 Carbonic anhydrase 1 1
52. Evaluation Hit selection blr Biacore S51 Eyaluation File Edit View Evaluation Tools Window Help Evaluation Explorer P salSensoreram 2D Quality control plots 44 Sensorgram Fc 1 Spot 1 sg Sensorgram Buffer i Bindin to reference spot Controls Bindina levels ot Controls Binding stability Result plots Binding levels ES Bindina lewels yersus stability Bindina stability early Binding stability late i Hesult Table Sensorqram Displays sensararams for analusis cycles either singly or in overlay plots Cywcles that are excluded from the evaluation are showr with a broken line in the sensorgram Reference Fc 1 Spal r Menu bar Evaluation Solvent correction shSolyent Correction Sensorgram RU Fc 1 Spot 2 r 50 7235 Cycle 40 7235 30 7235 20 7235 10 7235 0 72353 Response Act Ref 9 27547 Zoom lock 19 2755 29 2755 39 2755 2213 1713 1213 713 213 287 787 1287 Response Ref RU Calculate LClose 2 Calculate
53. Ea NI ceeie Lurye SarieOiOiah Foe m nnlm1 FJ Interactive Run qi1EOD i100 1 ammn Ll Interactive Run 395nn nm Spnt FRUG Tme ESHesP Slope Basehns Rashesp FL SgrthIar Fc 1 Spot 1 Time 4s Response AQT RU Fla ces 1 Temperatue 5 00 C Runming mteractree ul de 377 3 Inieck Stat RZE3 sqlion 2 flow DMSD 437 3 Iniect Reay 3D ul HDS washr Shat End Run TR DMSO Tool bar View Reference line RU Intaractive Run 40100 40000 39900 39800 39700 39500 39500 39400 33300 3J200 a ms 33100 1 0 100 200 300 400 500 600 700 800 900 Biacore S51 Instrument Handbook 46 3 DMSO Tool bar Edit AddReport Point Add Report Point Sensnrgram Fc 1 Spote Timm I5man wmdowl5
54. 3 Solvent correction Run solvent correction cycles Run Correction cycle every_ saqmple cycles 20 Number of correction points Biacore S51 Instrument Handbook 52 3 Next gt Cortact hi iissocaation WE 300 fe Etra Wash fher pectior vth DS Duplicate samcles Sabilee ahon fi laLllzaliri tm Help Sample injection Contact time Dissociation time Extra wash after injection with Duplicate samples Stabilization period Stabilization time Next gt Biacore S51 Instrument Handbook tore 50 DMSO 1 2 Sam
55. BR 1002 09 type 2 BR 1004 11 4 ml 2 ml BR 1002 14 type 2 BR 1004 11 2 ml 96well 384well Bidcore S51 Instrument Handbook 12 1 1 2 5 Tools gt Eject Rack Tool bar Wizard Wizard Rock Positions _ ek CHIT TK mW E 30 Sample compartment port
56. 79 Cle 2B 27 5 4 28 2d 42 41 40 2n 21 31 3n 45 dE Bd 3 E83 24 25 19 105 18 10E 15 14 51 52 5E FE EE Es EE Yes Yes Es Es Es Es Es Es Es Es Es Es ES ES ES Es es es Tes Yes Es Es Es Es Es Es es Cywcle Tyhes Fale Hames t ul tila t ul amle Same SITIEIE SITIEIIE SITIEIE ITIIE 5ITIIE 5ITIIE 5ITIIE 5ITIIE 5ITIIE SITIE SITIE Same SITIE STIE IIE IE TIE TIE TEIE TIE TIE SITIEIE SITIIE SaITIEIE LT R1a5 R1aE R1H3 R1H3 R1 R1 R118 R1 18 R1 amp 11 11 4 1 4 R1 R1 R1H1 FR1H1 FR1B1I 1 11 R110 1 FR1 R13 R1h1 FR1 3 R1D1 1 1 1 1 R1H R1H R1HS Ei E E 15 15 12 12 11 13 13 5 11 5 3 Irdnhrnfen 3 Iridnhrnfen 1 1 1E 1E 1 i SE 4 SB 4 5 G tst tta Sd S44 S44 DWR DWR 3 3 pullWl pullWl pull BI a1 3 BI 1 3 538 538 Ss Ssh Ine FE nmr Pt 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H 3H H LL LL LL 3H H 3H H 3H H 3H H 3H H 3H H 3H H
57. 48H Ad whoumd Lalrulation Compourt ldeemnaag 0 0 0 0 0 campoundconcenhabon On mm nr Impafl diclofenac IE Spot 1 Hosa 5BBg 5 diclofenac k Spot 2 mlsP CC Help hK Lancal OK Hm hound Tahles Riormal Blinod WL Close Biacore S51 Instrument Handbook 111 0 Evaluation Explorer bound tables miliiira am Lb 1 Ewaluation FE Edt Wiew Evakhaton TnnE Wndow Hm ET Dualty nnhial nos ImHirrg In reference 5 Coral Ering lsvel Lcrinls Erinn Hi Re po Enmc th ms Bindrg he 1 5 iciotanac IE Spot 11 FEA Bindrmg hel wei 1 EES clohanac kK Spot 3 AGP 1 EE Toatal haund 8 Bindieg BEIHIHIEIHI a ss bound mhinrrrial lood alue EE 10 0 2 374e 5 diclofenac K Spot 1 HSA diclofenac HI 2 5Ede 5 diclofenac KSpot 2 AGP Tuoital mmm Biqcore S51 Instrument Handbook 112 3
58. Tools Set Temperature i et Tenmheratures nalysss Temperaturec FC Hw K wl 4 45 C OK 25C Biacore S51 Instrument Handbook 1 9 1 2 4 SPR Normalize Tools More Tools 11 Hiacore S551 Conktrol software on CO File Edt Wiew Run Tools Windoww Help Evaluation Cr E WInn Caculator Charge All SmluHions hu dan Standby top Sianby Eject Rack Eject Chip Temperature Preferences 0 Maintenance Tools PR Desnrh Desob and Sanitize Superclean Tes Tnnls EN rmalize i System Check FE Service Tnals ew Flow 5ustem Wyash 5nltware Probler Fieport Mnrmalzes the sinnal in flom cal Tnlal run me 1s ahnut 11 minutes NOTE hn nnt narmalize with Ihe maintenance chip LastHunTime 17 lan 5 18 ue ce Normalize Start Biacore S51 Instrument Handbook 10 1 Mormalize x This nrncedur homalizes the detector sighal Total
59. be lt Biqcore S51 Instrument Handbook 3 51 En I Detection Flow cell Spot Reference spotduringrun Flow rate Flow rate Conditioning Run sensor chip conditioning Conditioning solution Contact time Number of injections Run startup cycles Sample solution Number of cycles
60. Result Table Binding level Resp RU gt 100 RU Da R1aE R1aE 1 3 1 3 11 7 R1 1 L112 RR1 18 RR1 11 1 R1 1 R1 8 15 15 7 7 12 12 11 8 8 78 Biacore S51 Instrument Handbook SR 4 4 Be 4 rata pci 4 4 3H 3H LL DL 3H 3H 3H 3H 3H JH DL DL DL leWels Fc 1 era 1 F ETF Fesp RU RU ENT ENT Mi TO 9d 1 4 5 d 3 Sg 1 5 1 2 5H 8 3 81 3 2 4 Hit selection blr Biacore S51 Eyaluation File Edit View Evaluation Tools Windaw Help Eyaluation Explorer wlSensorgram Fo Sensorg yn Carry ower oN Binding to reference spot Py Controls Binding levels Result plots Bindina levels Sensorqram Displays sensararams for analusis cucles either singly or in overlay plots ucles that are excluded from the evaluation are showr with a broken line in the sensorgram Helference Fc 1 Spot r wleeei
61. Temperature Sensor chip port 9 sample compartment port Waste compartment door Buffer compartment door Biacore S51 Instrument Handbook 2 1 6 2 S1 S2 S3 S4 S3 S54 OO
62. Biqcore S51 Control software Menu bar File Print Print Resuls Piinter hp cieskjet S90cs3 a System infnhmiahnh oer rte HInHmrs Eventlog Filo page elup _Moe gt gt a cpcles Moe gt Results OK Biacore S51 Evaluation software Menu bar File Print Finnter Name tahuis Haadm Type hp deski 930c sermes We USBOOT Camment Baselre Got Al Bd to refenerce spot Corntols Binding keweks Deselect Al Lonlrnls Innn stabihy Birm lgymls et Crent Biredimm lewels ymrsus stabilty Bm stabilhy e Bm W ate EE Et Result Table compettion Pt Sgs Ho setting avallahle He _ cereat OK Sensorgram Biacore S51 Instrument Handbook 4 113 4 Tools gt More Tools Maintenance Tools
63. REE i Biacore S51 Instrument Handbook 1 13 Rack Tray Eiected Hack Tray Ejected lck OF to rahkmri the rack haw to the me Enrnhhmerr Sample CnrmDarlrraf grmmaf uf 5 IEurmerWl He Sqmple compartment port Sample compartment temperature Rark Tray Ejecked Click OK to rahumm Fm rack tray to the am COMpar beert Sample compartrmer ermperaturne SC IEurrer Sample compartment port 60 OK A i Eh rack tray nsetbed im te irestniment It le recomemerded that you imrmmw ha rack tray so that it can be prepared ha starting a new FU Eject Rack
64. Startup Startup Startup Startup Startup Startup Startup Startup Startup Startup Control Sample Control Sample SaImhle Sarmple Sample Sarmple Sample Sample SaImhle Sarmple Control Sample Cnrtral Sample SaImhle SaImhle SaImhle Sample SaImhle SaImhle SaImhle 5 7 7 8 Sample Rias a 8 Control Sample IR1D1n Control Sample IR1D1n Sarmple R149 5 DMSO 5 DMSO 9 Helference Fc 1 Spo r RU EE EE ep RD Resp Ry moap RD lhesp RU lme 12 4 4 B co CO mm gt sl JE ool t rm mn lnlnlelo nD No CO 7 2 1 2 5 2 5 2 4 1 9 2 0 9 1 8 0 2 12 1 8 11 1 0 n 9 1 1 1 11 a 1 1 2 1 3 1 5 2 19 1 3 1 1 2 1 3 1 3 1 5 1 4 1 5 1 5 Biqcore S51 Instrument Handbook 76 3 Menu bar Data gt Table Columns WL Table nlummns General Plats Markers Statistics Bankings Characterization Sample Data Columrs Cucle Tupe Fosition Sample Mame da La Conc phd Data Corections 5nl
65. 100 RU Da Evaluation Explorer Binding levels Biacore S51 Instrument Handbook 3 67 Binding levels Evaluation Explorer Binding levels versus stability nigh affinity low dissociation rate binder 1n0 ERUDa c 1B c amp ES ad 5 gt zzZ EC c E SE EE S Gs lt 8 as in 11 73 Digitoxin TZ Digitoxin s854 tg Pydn de low dffinity high dissociation rate binder 9 Azosilfamide Soin 0 Mosulfamide 77 Eyrosemide RL Es Azosllfamide 5 i cv Tr PS 9 g 0 8 dS 8 1 Et E Qi 0 1 ga 3 no binder 3 a 68 8 im Ss 3 3 Ln mw Wn A lt b5
66. 50 ml Desorb and Sanitize Start Desorh and Sanitize x Th procedue romenes adsorbed mabenal ard dsiects te fom system The re 1 ded itn ous sheps Total ur hme sbit nne hnum Inllnwued bm COmimgahidad andEmi Hime hnums NHTE se the Markeriarce Chep For this procecdhae The sufacg nr other sensor chips Ma EE Chamaged by IE olutiorr us Dio nat uh thes piocett ne balnm a TC in nol sho this procediune er tm Next gt ES Desorb nd Ganitize x Rumed solutiorrs lfrom Mameriarrcg Ft Gees 5 BI solukiem 1 ahoul 5 sm snlutiom ahnmul sinfeptarrt snluhom aouW 3 mm Next gt Desorb and Sanitize lep Placa Bdmsnh 5nluhm in the readent cnmpaitmiBnt ared irset mp inlet ui s Buffer compartment door BIAdesorb Solution1 6 Start 1 2 Biacore S51 Instrument Handbook 4 117 Er le WiDe he LTp lg huhinHS wath a mata ISLUM Pla
67. Back tart Llose iacore S51 Control Software on COMI binding condition Sensorgra TP Ed Fie Edit View Run Tools Window Help 2 slesef i i Furosemi ide 10 pM 50 pM c Spol APROG Tme AbsResp Slope Baselne HeHHesp Id lr SmAct 16 0 3E555 005 Yes 0 baseline 1 1r SriAcg 4200 35529 004 YY 3 6 baseline vent jog bliacied Fc 1 Spot 1 r Time Ino Tine 816 0s Response 3666 2 RU DO Tnestamp 28 Mar 013 16 25 15 a RE Flow cell 1 emperaluse OU Temperature 25 00 C 00 Sample Temp Seling 25 00 Pow oek 1 Standby flow max remaining time 4 00 Days 00 Flow 30 pmin lt Biacore S51 Instrument Handbook 3 39 2 Bimding Conditions Results E _ eisnf la Sammlelnleclm werlaW Cycle Fa necticon gc mection En in En 1 10 uh Helb LCle enu Llpse Ko 2
68. Buffer compartment door 6 Start le Fac brl cnriainin Eh in th a readeht oompaitiment TH is He Fm hh mns Krom all pumgps This slsp ushes the puimps vth 7 harml 70 Start 5hutdnwm ntpp Fiamanwe thee hubings Irnm the mhhannl bottle grid alnw Ihem bo har Im the air This shep emnlies IP ut sd um uhin of lguid 70 Start 5hut lm gl 4 Plgasg wlhch off Khe imsTumgrl 8 NBN 295 Bidcore S51 Instrument Handbook 124 5 5 2
69. Fankings hegatiwe positive control A A ES Elys riEgEagtire nnsitiye cnritrnl 14H 12U E 100 E EE h Me I 8 Y 1 FE 5 8 2 4 nsiiye cnrtrnl nn SR a me 2n ml NE LE ned iv mW gH 1 1 12 1 1E 15 2 Cycle number Biacore S51 Instrument Handbook 72 3 RU Sensorgram Azosulfamide 50uM 3980 gt G D gt 3960 gl W EE 3940 5 3920 mm gt ox a GO NN 8 EE gt gt gt G Js a 3880 5 8 8B 8 OS MK 3860 0 100 200 300 400 500 600 700 800 Time Evaluation Explorer Quality control plots Hit selection blr Biacore S51 Eyaluation File Edit W
70. Hep Compound Compound concentration 10uM Import Biacore S51 Instrument Handbook 3 109 TH Import ED Yakue Lompound charactesizations 5Em B Finebhers Spot T infcamathon 5Paot 2 infomation na armnlei clnferrmc Ligart H5a Ligard AGP He K es Compound characterizations KD values Ko M OK Add bound Calculation Kp M Kp Origin Protein Prot Conc HSA 650 u M AGP 24uM SH Add mw bounmd Calculation Lompound ldiciemag campound canesnrahen in Im Impat diclofenac IE Spnl 1 Hep OF Cancel Biacore S51 Instrument Handbook 110 3 Import
71. NS ST OB SENE SSE x Oe 2 EM BY Sr OT EEO OCS Bs CE io IO io ET gt oa No MW WY Qi oo 2 RR 0 YA CN NN SM 1 0 10 20 30 40 50 60 Cycle number Staqrtup cycle Buffer Controls Binding levels binding_late baseline Result plots Controls B
72. Rmax 6 sec Biacore S51 Instrument Handbook 40 3 3 1 3 DMSO Interactive Run DMSO DMSO 500 1500RU Menu bar Run Run Wizard Tool bar Run Wizard Interactive Run Start 0 Run Wizard Sumface Preparation ImmahllEahnn Immnbllgahnn pH Scouhrng 0 Assay eyelnpmerW uaa Bdg LnrrHWinns Hegeneraliom Cordticns Fiot Hu saWs Hi 5mlerli
73. 5 1 3 Standby flow 3 Standby flow Tools Standby ny SS hl aarmum stardby hme E ms nmumi requeed hl wolume 345 imll dik _ Ese 7 Next gt Desorb and Sanitize 70 Tools Shutdown 5hutdawm This pncedure empties the fwe smshemm ar sh He the insIrumant The procedure amp dded to four teps Total um bome i hnut 11 minuhms HEHUIled cions lenieed vater 7 Hrarml Next gt Biacore S51 Instrument Handbook 5 123 5hut om _ x step 1 Flace a belle cnmlaimin deionized wiale mi the eagert compa tert ar nserl the inlel hubings hom a pumps This shep Fushes the whole Hc sphemm hi wuahe ahH IFmeri rnpiies le FL
74. pH Immobilization pH Scouting Immobilization pH Scouting pH CM5 pH J 0 5 pH 10 mM Immobilization pH Scouting pH
75. 30RU Rmax 30RU 40kDu 200Dq RL MWL Rmax IMWA MWL 40 000 50 200 1 10 000 10 000RU Rmax RU RU RU Rmax Rmax Rmax 100 Biacore S51 Instrument Handbook 18 2 2 1
76. IDeianized water e000000 6 e00000e OO e900090e e000900 8 e090000 9 OOOOOOOO Help Menu Eject Hack lt Back Start LClose Start Stqndby flow Biqcore S51 Instrument Handbook 4 119 4 2 Desorb and Saqnitize CM5 BR 1003 99 1 HBS N buffer BR 1003 69 1 BlAtest solution Tools gt More Tools almterarcg Tnn 9 Desorb Desnrh and Samilize Superclean Tgst Tnnls Noimaliee Semioe Flaw Smstem rash Fm 5nltwiarg Problerm Fieport Checks for age arnd tests pumps blarnk Imlgchinns and noise Total run rme is about 45 minutes NOTE This nrncgdurg should he run at FL with new Sehnsor Chip LME Hunninn Bulfer shnuld hs
77. Use Temaise Hit Selection Start fn Hit Selection Assay Format Direct binding Surface competition Surfqce competition Biacore S51 Instrument Handbook 3 49 Direct binding Next gt Hit Selection Iniection Sequence shhPLE I Sampls WAIT Stabilization period CALRRY OYER FF Carry ower control T amp Adaptive FN sussEhlalEll 1 2 St abatinn eind Regeneration se saito dative Nos ElElli 1 2 4 Capture Sample Regeneration Stabilization period Carry over control
78. 49050 49000 48950 8 48900 48850 48800 0 8 8 48750 0 2 48700 n 100 200 300 400 500 En Time s Serrsnigrsm Fc Spot 1 a Sooo Por not Select Bsnr rar LuryE hame Fc 1 5 nn Fc 5nnl r Fc 1 5nnl 1 r cnrr Fc Spot 2 Fc 1 5hnl T Fc 1 5hnt T En Fc 1 Spot r 8 Cancel Bidcore S51 Instrument Handbook 84 3 cyee 1 Select cucle Sample Name II1 Startup Bulfer Yes II2 Startup Bulfer Yes IU3 Startup Bulfer Yes II4 Startup Buifer Yes II5 Startup Buifer Yes II Salyent Correction Yes 07 Control Sample negative 5 DMSD Yes II8 Control Sample negative 5 DMSD Yes II3 Sample CBS amp Yes no Sample CBSA Yes 1 Sample phernytoin Yes ni2 Sample phenytoin Yes 3 Sample azosulfamide Yes ni4 Sample azosulfamide Yes is Sample warfarin Yes Sample warfarin Control Sample regaive DMSD Control Sample ample Salyent Correction Control Sample negative 5 DMSD Yes Control Sample negative 5 DMSD Yes
79. DMSO DMSO 5 DMSO DMSO 1 DMSO DMSO 1 DMSO 2 4 6 DMSO 4 DMSO 9600 ul 100 DMSO 400 ul 10000 pl 6 DMSO 9400 Ll 100 DMSO 600 p 10000 pl 3 4 lt 6 DMSO 8 ul 470 0 Z00 400 600 S00 1000 1200 1400 620 1400 12Z00 1000 o00 600 400 Z00 1400 _ 1400 1400 1400 1400 1400 1400 1400 DMSO DMSO DMSO DMSO
80. TL TI Bimrlimn lewels Fc 1 Spot 1 F torr ER Eo 7 Th 9d 1 Et 4 3 3 2 31 5 31 SH SH 3 1 31 3 3 3 8 8 4H 1 HI 3 5 nn 50RU Menu bar Data Filter Data 1 Filter Data Cucle Included Cucle Tupe Pasition Sample Name kd Da Lanc pkd Binding lewels Eaulumn Sensorgram Fc 1 Spot 1 r corr all all all all all Show Fiows Filter Show walues In range 5H sl il73eMa Biacore S51 Instrument Handbook 80 3 Column Binding levels Filter Show values in range 50 50RU Cyrle Tyhe Hamne TW Da nmr MI 1 res sample TS IT SO Tes t 4 YES t BTes amble t dTES t 1 res IT 4 Tes TI 1 YES ITIIE res t res AanMhle Des t 65
81. ba tw ers Id Baseline OK Interactive Run 39100 0 100 200 300 400 500 En0 700 800 900 Fc Spot APROG Tme Window AbsResp SD Slkpe Quakty Baseline HelHesp ld 5H4 0 a Ye 0 baseirne a173 7 31737 0 41 121 Dk RelResp 0 DMSO Biacore S51 Instrument Handbook 3 47 RU Intaractive Run 40000 1 IaRun 584 0 50 331737 0 41 021 Ok 1 IAHun 5230 50 400403 3 70 1 97 Ok 040 0 50 382897 310 166 0 Baseline RelResp Tool bar View Reference line 1 DMSO 1 200RU DMSO 500R
82. 200 ul 7mm 20 C 4C 200 ul 10mM 5 200ug ml 10mM pH 0 5 pH Immobilization pH Scouting pH pH4 0 10mM RU Biacore Da
83. Sumface Preparation 7 Immobileaton Immohilzalinn pH Scaulmgi Assay Dewelopmert FF Inkeraclya Rum Bdrm Corndtions Heneneraliomi Corndticns Plot Run ssaws Hit Selection Compound Characterization Tests the ebert of birhirg of ligard to the unacmyaherd sensor sutace urrHai differert buffer cnrditiors Optmizatiorn of He bulfar pH ts mecessay For efheiert immobilgzatior Immobilization pH Scouting Start Irmimabilization pH Scouting Buffer Selection 1 ml sacetate pH 5 BI TU mh cetate pH SO 1 ml cetalg H BI 1 ml cetate pH Next gt Add pH Remove Biqcore S51 Instrument Handbook 20 2 immobiltzation pH Scouting SENl x Flo ar cedection NU 3 LI and sethns Name Cankact ime min Fl hm iumin Gurace BEf ai This eernieralinn wl be peafhommed nr a the Er of the weand lutorr rd aH Hep ese Flow cell Spot 1
84. fAssay Dewelopmert FN Interacte Rum Bmdmn LondWinns Henenefahmm Cortions Piot Run Assars HI Sebertion 2 Lammnaurd Eharaclerigalian Parlnirns cnwalami mmmnhllcahnm of li nand nh he semsnr sulfacs The sa nf dfeer narrds can be immnhllca cr bclh datachnm spots Im the Fwu cl in a sgle ur Help Immobilization Stqrt rmmabiizatinn Setup msi chip Chip pe Is Flow cel Flow call 1 Flow call 2 Flow ll hgads y Ligand in spot 1 Chip type CM5 Flow cell Flow cell ligands spot Next gt Biacore S51 Instrument Handbook 2 27 EE Fiethod rirmcdbhizat on mn mw cell 1 lire on hn ar LA He Back Ket gt Coee spot Amine Surface Thiol Custom Methods Custom Methods
85. tatup 1 Buffer 1 BI YEs tatup 1 HuTffgF 1 2 Tres tartuph 1 Buffer hE YES tartuph 1 Buffer OD Yes tatup 1 HuffgF 1 es tatup 1 Buffer 1 131 YES Control SaImhlE IF1D1 Sa hkS 2 131 Tes Control sample R112 35 hd 14 Yes TIE RR1 1 1 53 2 4 15 YES ti RR1 1 1 53 3 TE YEs SaITRIE R1 2 5 3 1 3 1 es SaITIE R1 3 11 H 1 YE SaITRIE RR1 3 BD 4 3 1 YES SaITIB R13 EBD 4 3 2 es SaITRIE RR1 ta 3 2 res SITRIE R1 ti 1 b TEs Control samples 1R1D11 35 LMSO 1 Cnrtrnl SaImhle 1R1D11 535 DE 2 YS SaITIE RR1 54 3 4 SYEs SaITIE RR1 SH 3HH 4 Binding levels Menu bar Data SortData El art EE ta Binding levels Fc 1 Spot 1 r corr esp Em Descending Then hr Lnclg sehnding Descending Then by rone scehnding Descending Sort by Binding levels Descending OK Biacore S51 Instrument Handbook 3
86. zaamLack lt Back Net gt Eose 0 M Zero conc subtraction 0 M View average 0 M Zero conc subtraction 0 M 0 M Zero conc subtraction 0 M Zero conc subtraction All compounds 0 M
87. 30 Sample CBS amp Yes 31 Sample CBS amp Yes 32 Sample phenytoin Yes 33 Sample phenwtain Yes 34 Sample azosulfamide Yes 35 Sample azosulfamide Yes D3E 5 Samp warfarin Yes no Cl rt 1 Ma cycle Ctr cycle OK mlSensnrgram kk GETsnT ra FE nt r corr RU Sensorgram SLI 5 HH ReshnurisE 35 UI Time Biacore S51 Instrument Handbook 3 85 p5 25Bn 2 i 3 ia a e TP BE ri nir 325 EC EO 1 1 3 OD Sh EO Tire menu bar View Adjust sensorgram Gansnrgram diustment TS Zenatstart of cucle Noadjustment i Zero atreport nih baseline baseline 5 Fern atinectior Zero atinectior Sample IIBChnr start bd ample IIBChinr start bd Lahc X axis Zero at report point baseline
88. 99 UIJBJBA 9 SDIUUBJITSOZB 9G SDIUUBJITSOZB GG lt UIO1AUSUO g lt ulolAusyd 6G lt VSdO 19 OS G SABDeu 0G OSWQ 8 SAnebeu 6b UIXO1BID UIXO1 BIp 9b lt 9pIWssoln Gh lt 9DIUU9SOJn gt U9XO OBU 6 USXOJdBU 9SDIUUBIIUEJITS 9SDIUUBIIUBJITS 0t OS G SAnBDeu 66 OS 9 9AlB69u 9E UIJBJBW 6 UIJBJJBW 96 SDIUUBJITSOZB G6 lt SDIUUBJITSOZB 6 lt UIO1AuSUd e6 lt UIO AU9UO 6 lt VSdO Ie lt VSdO 08 OS G 9Alebau 62 OS G 9ABD9u 9Z UIXOIDID 92 UIXO1IDID GZ gt SDIUJSSO SDIUUSSOJn 6 USXOJdBU 22 USXOJdBU Lg SDIUUBIIUEJITS 02 CarrV OVer SDIUUBIIUBJITS 6L OSMQ G SAnBD8u 8Le OSIAQ G anebau L UIJBJBAW 91 UIJBJBA GL 9SDIUUBJITSOZB 9DIUUBJITSOZB 6L lt UIO1AUSUd lt UIO1AU8SUd LL lt VSd O 0L lt VSdO 6 OSAQ G 9Ane6eu 9 OSAQ G 9Ane5eu 19jjng G x 19jjng 19jjng x 19jjng 2x 19jjng L x 5 6 TN o 5 NN lt gt 91B DUuIDUId OO ssuodssy Sp 40 50 60 30 Cycle number Buffer 20 10
89. Biacore S51 Instrument Handbook 62 3 12 1294 Cucl 2 12943 8 7 87057 1 17 87056 Znoam lock 27 87HB 4 37 8705 8089 7089 5089 5089 4089 3089 2089 1089 89 Response Ref RU Help Menu Calculale Close Calculate EE Sample injection Em gnu Inrludg Crles ith Exrlude Aall Crles WiEh Fl Report Pnints Outside Correction Rari g Report Foints Inside Exxtrannlatinni Raring Continue Biacore S51 Instrument Handbook 3 63 Response Refl 10
90. Fit Series 5 esorb snluhinn 1 BIadgsnrh snluhinn 2 Next gt 16 mm BIAdesorb solution1 BIAdesorb solution2 1250u Desnrh Rark Positions i ET ET 7 128n eh OS s 1 RET IB2 Deionizedwater 51 0060600906 Hglp gnu Eject Hack lt Back Hart Llasg Start Stqndby flow 3 4 Biqcore S51 Instrument Handbook 116 4 Desorb and Sanitize BIA BIAdesorb solution 1 0 5 SDS 4C BIAdesorb solution 2 50 mM Gly NaOH pH 9 5 BIAdisinfectant solution 3 8 ml
91. Flow rate Flow rate 10 30 90 ul min 30ul min Sample Name Contact time 60 Dissociation time 300 Concentration first injection Concentration second injection 10 50 M ks Kp 10 0 1 1uM Next gt Binding Conditions Swstem Freparations Flow snstEm preparation None all solutions Nomalize detector nalusis Emrahure Next gt Biacore S51 Instrument Handbook 38 3 Ginding Rack Positions EH TE CO IFurnsarmida SU uh Hep henu 2 1
92. HES LastHunTimg 1EJuHH3 TE _ Hep sn cmcm Test Tools System Check Start EE sbi Check This pancedure should be run at TL wilh a new 5erist Chp LM and wath HES N as um bulfmr Choose Close you need to change lhe sereor chp sl the ternpetature or charge rui bulfer Fieagent pu Es 3 rr ar EC HI B Riefractometer painrmawice and llnw cel leakage SS PU Cr ihn a sacldiessing EE Neots Tests WIhe peristaltic plumt In neder rd thal a sameie inechom wi buffe Iam the Heanerl supply block t a ii hL Biacore S51 Instrument Handbook 120 4 Next gt I System Check Rack Position Reagent Rack 2 FF eeeeulveLeul cemew BvWer IS3 Deionizedwatermhh5sopopp water s4 IDeienizedwatermn OO OO OOO Help Henu Eiect Hack lt Back Start Close Series S BlAtest solution Sstart TIE 3nzn5 ui Wy 3nz18 30218 30218 oanzig gal 5Wne1213H 30224 nz1zl3P nanzz4 He1213P1 I Mg Matwork F
93. REGEN 7 Hegeneratiort Lise two solutions ddress spots separately 1 2 7 AAT hw Stabilization period _ Hep lt Back Eoee Biacore S51 Instrument Handbook 58 3 Capture Address spots separately 1 2 1 2 Capture stabilization period Next gt 2 1 Wizard Hit Selection Ihjection Paraimeters Ligard capture separately addiessed spots Ligarnd
94. ruh time is about 11 it Hgnulred snlulinn fro aintenance Kil Gerles 5 BI nanrmalzinn snluhinn Next gt Mormalize Rark Pnsitinns Position Vol ph Content Rg FE T1550 Bnormalization solution IS1 olBuler Hganght Hack 2 Bufer ho WWW IB2 Deionizedwatem oo Help gnu Eject Hack lt Back Start Llnse BIA BIAnormalizing solution 150 u 7mm 11 Start Normqlize Close Standby flow Biqcore S51 Instrument Handbook 1 11 7mm BR 1002 12 type 3 BR 1005 02 800 ul 16mm
95. 50 Help Zoom Lock Bac Next gt Close Biacore S51 Instrument Handbook 94 3 Compound Sensorgram Reference subtract qnd corrected Reference subtract Reference subtract qnd corrected Include Fc 1 Spot 1 r corr Fc 1 Spot 2 r corr Pl dd Compound Characterization FE rmrmobilization properties For Fc 1 Spot 1 rmrnobilizatinn properties for Fc 1 Spot 2 rmmmobilized la caboric anhudrasg mrnobilized Ii ard Immahilzalinn lewel HL Immnhillgalinn lewel HI Immnhilgalinn date DS 1 dA CE mrmobilization date Eomment Comment Fr 1 Spot 1 r corr Help gnnm Lack Clase Next gt Biacore S51 Instrument Handbook 3 95 m dd Compound Characterization grn CnhC subtractior Fc 1 Spot 2 T En FED Cone suhlractinn Fc 1 Spot r corr Encle dE zansulfamide compounds Wiem awerage Help
96. 9U 6Z OS G AB59u 8Z UIXO1IDID 92 gt UIXO1IDID G2 2 9DIUU9SOJ 9DIUU9SOJT 6 USXOJdBU 2 USXOJdBU 1 9DIUUBIIUBJITS 0 lt 9DIUUBIIUBJITS 6L lt OS 9 Ale6eu 8L OS G AneDeu L UIJBJJBM 9L UIJBJBW GL lt 1 1 Baseline 9DIUUBJITSOZB UIO AU9Ud 2 UIO AUSUd 1 VS 01 2 VSdO 6 OS G 9AlBD8u 9 OS G 9AneBeu 19jjng G J9jjn 48580 48570 48560 48550 48540 RU Sull9SBd suodssy 48530 48520 40 50 60 30 Cycle number 20 10 ass mi Ss 7 2 W RU 10 Carry 0over co bindin g_late co_baseline OS 9 D U U9 9 OS G Anebau 6G UJBJBA
97. E aLiuE Control Control Furnsenide xx Biacore S51 Instrument Handbook 3 55 Sample order As entered Randomized Next gt Hit Selection Swstem Freparations FInwi sustEn preparation Hglresh How sustEm Marmalce detector nalysis lemperature Hack Close Next gt Bidcore S51 Instrument Handbook 56 3 Hit selection Rack Positions 1 1 1 OCOC C LOLOL OIGI OIGIS R 23 EN mM 1 NM RMS EE Sulanlame 5 phd _BB_ pm EE Contral Nastia control 0 phd Control Manatma control OD phd RIAll BE Control ConroalFurosemide 50 nM RLA12 Cantrol Control Furosemide 50 ud EE correction solution 1 predip Correction solution 2 Solvant cnrrectinn solution 5 RDG5 6 Mail EG correction solution EE Es TE R2FE 0 BIAnormalizinc s
98. E h 2 TE EBIIEEHGTIBEII s amp MIFLE 7 Sample REGEN HEnenralinn ghm snlulinis Adaptive bmmemee ee Aaddress spots separately 1 lt lw SGtabilization period wv Adaplive 4 A TY es ICARRY 0V ER ki le Cary ower cortrol cee Adaptive Next gt 2 1 Wizard Hit Selection daptieity Parameters Farfnrm regerneratiorn i the relatiwe response after dissnciation enceeds HL Im spot 1 T or IRH mm spot 2 awirmum humher of cnrrsecuhyE TBHETIET In IIECH TS E Fepeat the stabilization period the response Chan Es more thar RD s mm spot Tr or IRHLU HU Im spot 2 F awirmum humher of stra stabilization perinds Farfnrm carr nwaT CHIrnl if the relatiwe response the ehd of the imectinh enceeds HLI Im spot 1 T or RT In spot 2 r a des wirmum humher of Earru nwEr controls 2 1 Biacore S51 Instrument Handbook 3 61 3 2 2
99. GE Healthcare Biacore BidcoreS51 Instrurment Handbook gt RUNRRUYTD GG l ed EE KKKKK 1 l NN 1 1 1 1 Mui 1 jsds2 Sooo i 3 2 4 1 2 1 Insert Chip eee 4 1 2 2 Change All Solutions 7 2 8 oS Aid 8 1 2 4 SPR Normalize lr 9 1 2 5 NN 12 ls2c6 Standby Si 14 2 8 15 2 1 pH Immobilization pH Scouting 18 2 2 Immobilization NYNNNNIINN ll 26 3 1 30 3 1 ee 35 3 1 1 Change All Solutions 35 3 1 2 Binding Conditions es 36 3 1 3 DMSO Interactive RUn ee 40 3 2 Hi
100. I 25000 Ii Sensorgram window 15000 10000 SI Dn 1 1 100 200 300 400 SoD BI Time S Fc Spot A amp APROG Time bsHesp Slope Baseline HalHesn Id SamplesTgpe Name Pos Mw Conc 2 1 sample 9 37248 3 22 Yes baseline Startup cucle Buffer 51 2 2 sample 25 9 35EB5 4 D2D Yes baseline Startup cucle Buffer 51 2 sample 25 9 35487 1 D0 20 Yes 0 baseline Startup cycle Buffer 51 2g 1r sample 259 17B1 2 DO2Z Yes haseline Startup cucle Buffer 51 2 2 sample 25 9 1743 DOD Yes DD baseline Startup cucle Buffer 51 Reportpoint table 2 1 sample 50 9 3724E B HH No 1 7 bindina_early Startup cucle Buffer 51 2 2 sample 50 9 35EE5 3 OD 02 No 1 2 binding_early Startup cucle Buffer 51 Ewent log Subtracted Fc 2 Spot 2 r Time Infg Time 5UU Us Hesponse 179 3 RU HH Timestamp 02 04 04 143 49 07 DO SetTemperature 25 00IC Flow cell DO Temperature 25 000IC E Ii Sample Temp Setting 25 C Eve ntlog ta ble 8 00 Fowcek2 nstumento Stqtus WITOOW nn Flow Emin Menu bor Tool bar Sensorgram window Report point table Eventlog window Sensorgrqm window X
101. Spot 1 RU so000 4so00 40000 35000 Response 2 3DOOD 25000 0 500 1000 1500 2000 2500 Time Fc Spot APROG Time Abshesp Slope Baseine RelHesp ld Ligsnd Ligandyol LigandFlow 1 1 Amine_Spetl 71 0 29491 2 018 Yes 0 Bassaine 120 1 Amine_Spo1 8340 29711 5 006 No 220 3 Actwahon CA 120 10 34275 9 1 45 No 47857 Ligand CA 120 10 3551 5 Immobiced CA 120 10 1 1 1 Amine_Spcotl 1532 0 1 1 Amine_Spot1 X1670 330428 005 No 231 0 s 33040 7 RU Not connected to inshumeri Im of CA Results Sensor chip CMS Flow cell 1 TE nt fime 12 1 min Hoe 10 Respornses 1 hmuhd SES 4 RnTis fimals 3551E RU Response1 bound RU Response2lfinal Response2 finqll Bidcore S51 Instrument Handbook 30 3 3
102. Tmormmrilh lt lEHERSOEEPTommn Sample positioning Sample positioning Sample positioning Manual Automatic wertical Help Cancel Help Cancel Help Cancel Manual Automatic vertical Automatic horizontal Biacore S51 Instrument Handbook 54 3 Next gt Next gt HIt Selertinn Control sample Table ND FR Menu Hack Mei las Cycles Fred Cycle Freq Next gt Hit 5elertinn Cycle Editnr I f As gntered Randomized ESEBPEOD 0C16 re re startup Cucle Startup Cyele tartup Cucle Startup Cycle Soluent Correctiaon Heqatiue control Control Furosemide Digitoxin Warfarin Ha Fhenuytoin sulfanilamide i FU se zosulfFanide H
103. U DMSO DMSO Bidcore S51 Instrument Handbook 48 3 3 2 Hit Selection wizard 3 2 1 Menu bar Run Run Wizard Tool bar 2 Run Wizard Sumface Preparation x Immnbllgahnn Imimnhilgahnm pH ScEnulmn 1 Assay hewelnpmefW FF Iniearacte Rum Fx Brdg Londtions Henehefahmm rw ICs FF Piot Ru Assars HE Sahertor Campourd Characherizalion Perhoams rcuhrm aralyte bindng assars nr sampkes Im dB nr Bel microplates cluding pushwe ad nanalive cnnhnls and repeated nlant Corechon cpckes se thes wigard for secorday scieerirg ealy ADME ed thet oteirn bg assas Help
104. ca BLAdesnh 5nluhor 2 In the reagent compaltman ard Insert ll Pammp inlet uri ph IF Sat pom BIAdesorb Solution 2 Start 2 Sqnitize pesarb LIL GTiTillHFT ln you wart to sartiee he sysbem int sarniize at as ge Sanitize Next gt Desnrh and Sanitize _ nle 3 wipe lhe um Inlgt im Wtfh mal sh Place 43 ml diuted Basintectant Soluticor iT he Bf comparimert arid Wiiell al um riet tuberngs Sanitize Start 30 Saqnitize Sanitize Step4 Start Desorb and Sanitize The gsnrh mr 5 IOCgHuTa completead a Be ia stem tn rum in stardby mode inr at least 4 hnurs bafore Deriormwm TL Close
105. dg H27 5m 3 Sulfanlarmida 1 San amide 13 sulfanilamide 1 ulfanilamide 1 Sulfanilamide Sulfanilamide Sulfanilarmide 18 Sulfanilamide 119 Sulfanilarmide Sulfanilamide Sample mcioplate Pe 35 wellMicoplate 7 5 ample pennim Manual Reagent rack type HeagamHack2 Help nu Back Ned Close 1 0 1 duplicate_ n 2 Next gt Next gt Bidcore S51 Instrument Handbook 92 3 Cormpoaund LhasarartEriraLinm L rlE E Lnr IX Startup Sart cle Increasing i TS artup Cycle hs entered Startup Cycle RS Startup Cycle Radomeed th Soluent Correction nr Kh 7 Furosenid a 1 ee C 1 Sulfanilamide u rT Soluent Correction Ln ky 28 EBSA C 3H Azosulfamide hd Soluent Correction Few jsbem Pierator None Change schiions R
106. e well custom order Same well consecutive cycles 30 120 stobilization period 53 Ht Selection Sample Table Digitoxin 7E5 aa R1gg 3UB 1 3 a ehoin R14 2 OM R12S 172 RT ai1 TP R17 543 HIAznsulfamida R1eg 1 eames Mciopiake ips dE wall NiEinplamle 5 ample positionirg Marmual Beagent rack type ReagentAack2 r Hep Menu lt Back New 8 Sampls Fosibioning Import Sample Data nHHiEdit User DeFined nlummns Lelete Selected 5amnlers Insert samplets Sample Positioning EPRERYTEPTSCTTTl lt lENERTTEP
107. efesh few system Change all solutions Next gt 2 1 Biacore S51 Instrument Handbook 3 93 3 3 2 3 3 2 Menu bar Evaluation Compound Characterizations HHCnmpnund Characterizations Lnrnnnurd haraclerigahinns Hame Compound curwes Aetive SAAS Edi glete LT iky ll DPiyala LlInse Add Add Compound Characterization x Compound zsulfamide Sensogam Reference subtacted andcorected Include Fc 1 Spot 1 r carr v Include Fc 1 Spot 2 r carr Immobilization properties for Fc 1 Spot 1 Immoabilizalion properties for Fc 1 Spot 2 Immobilized ligand carbonic anhydrase lImmaobilized ligand lmrmobilization lewel 4281 RU Immnhbilizalian lewel HL Immoabiizalion date 2003 03 405 18 44 08 Irmmobilization date Comment Comment Fc 1 Spot 1 r corr Fc 1 Spot 2 r corr 100 150 200 250 10H 50 SH 100 150 200 250 300 3
108. ent Handbook 70 3 Add SH AHH boundary positive control Hesponse 33 RU Help Larc Name Response OK Ranking plot 33RU positive control Add Ranking x Name vs negative positive control Description Sensorgram Fe 1 Spot 1 r corr Beport Point binding_late Ranking boundaries Control samples 1 0 lt View average values View al walues ti rtral 33 6 rme Edit Control Sample Iconc mM Aug i D 4 Delele Necgative i Control Furosemicde SD 3 Hankina plot positive control gt 8 10 Cycle number Allow ingraph editing of boundary yalues Snrt by cycle Number Help IK Cancel OK Biacore S51 Instrument Handbook 3 71 Add Close Evaluation Explorer Ranking
109. es Select Ehsnr rar Fc 1 5 nnl Fc 1 5nnl r Fc 1 nl r cnrr Fc Spot 2 Fc 1 5nnl T Fc 1 GSpot 2 T Corr Fr 1 Spot r Cancel Fc 1 Spot 1 1 1 Fc 1 Spot 1 r 1 1 Fc 1 Spot 1 r corr Fc 1 Spot 1 r OK mlSensnrgram a 4 Cucle 9 bz EE nnm Lanck RU Sensorgram CBSA UM HH dO 4 ResHUTSE ar GE SI Tirme Biacore S51 Instrument Handbook 3 83 menu bar View Report points Marker and Id 3 44 Cucle 34 4 Sensorgram Fc 1 Spo 1 Zoom Lock RU Sensorgram CBSA 25IM 49150 49100 a g
110. etic Data Fc 2 Spot 1 r corr x Hint Dptions The Check Kinetic Data tool compares simulated sensorgrams eraged esrperimerta residuals Black original fitted curyes iuides Blue rate constants increased by modification factor M Red rate constants decreased by modification factor M You should reject the rate constants if changing the factor M does _ Compare to hot significantly affec the red or blue curyes 6 Sensararam See Help for more details Hesiduals Iniection stark stap easeeeeeeseeeeeseeeeeseeseeseeseeseeseeseeseeseeseeseeseeseeseeseeseeseesee I Zoom Lock 5D SD 100 150 200 250 300 Time s Decision Helectrale constants Help Cancel Modification faqcter M LCheck Kinetic Data Fc 2 Spot 1 r corr Hint Dptions The Check Kinelic Data tool compares simulated sensorgrams Yeraded EZ it resid Black original fitted curyes it quidgs Blue rate constants increased by modification actar M Hed rate constants decreased by modification factar M You should reject the rate constants if changing he factor M does Compare to hot significantly affect the red or blue curyes 8 Sensararam See Help for mare details Besiduals Iniection start stop Modification factor M pesssssssssssssssssssesesssesssesssseeessesessssesssssssssssessssssssesesesesessese
111. iew Evaluation Tools Window Help Evaluation Explorer salSensorgram duiy Corel plos 8 pee Sensorgram Buffer Bindina to reference spot Contrals Binding lewels i e EsENT plots 3 Binding levels a Binding levels yersus stability FT Binding stability early Binding stability lala i Hesult Table Sensorqram Displas sensorgrams for analysis cpcles either singly or in overlay plots Cycles that are excluded from the evaluation are shown with a broken line in the sensorgram Reference Fc 1 Spot r Biqcore S51 Instrument Handbook 7 3 3 Baseline baseline OSNU 69 SAIPUGU 09 OS G 9AlB59u 6G e UIJBJBA 9G UIJBJBA G 9DIUUBJInSOZB 9G lt 9DIUUBJITSOZB GG o UIO AUSUd 9 lt osna 9 69 9AlB6eU 09 OS g SAne5eu 6 ee UIXO1BID UiXO IDID 9 9DIUUSSOT G lt 9DIUU9SO USXO dBU C USXOJdBU lt SDIUUEIIUEJITS SDIUUEIIUEJIITTS 0 OS 9 9AIBD9U 66 OS G 9AlBD9u 86 UIJBJBAW 6 UlJBJJBW 96 lt 9DIUUBJITSOZB GE lt 9DIUUBJITSOZB 6 o UIO AUSUd CO lt UIO AUSUd Z6 gt VSdO IS lt VSdO 06 OS G 9ABD
112. inding stability stobility_early bdseline Controls Binding levels Biacore S51 Instrument Handbook Evaluation Explorer Result Table RU 1 Hit selection blr Biacore S51 Eyaluation 75 File Edit View Evaluation Tools Window Help Evaluation Explorer Sensarqram Quality control plots Baseline 1 Carry over la Binding to reference spot Controls Binding levels Controls Binding stability Fiesult plots Bindina levels A Binding levels versus stability Bo Bindina stability early EE EO i hesut Table Sensorgram Displays sensorgrams for analysis cycles either singly or in overlay plots Cycles that are excluded from the evaluation are shown with a broken line in the sensorgram slSensoreram 44 Sensorgram Fc 1 Spot 1 Sensorgram Buffer Evaluation Explorer Result Table de
113. nlutinn 70 mm He Mean c Hack Sarl Close 2 1 Biacore S51 Instrument Handbook 3 57 Wizard N 5 23000 22500 22000 21500 21000 20500 20000 19500 1 19000 18500 0 200 400 600 800 1000 1200 1400 Hit Gelection Ihjection Sequerhnce TT I Capture stabilization period hhPLE Sample
114. nn Compourd haraclerigalinn Enablkes userspeciied mpection of samples nw the sumfaca Use this wizard inr exploratory tests such as checking mm range of the solwent CTCWTBC ho NHUTE Temnlales Fon Interachme Run canrnol be sa Hm ueremwss sw cw Ei 1nterariye Run et _ x Flow Flow rate mn _ Dieleclm Fipw em sn Helh Flow rate 30 ul min Flow cell Spot Next gt Biacore S51 Instrument Handbook 41 Help lt Hack New gt Closs 5 DMSO solution 1 high DMSO 6 DMSO solution 2 low DMSO 4 DMSO Next gt MInteractive Run Sat PTE FI syshem fBDFalmm Nene Change rR a EY ND RN UD EE HN HN IL PT ET TT Tt Hompalize deleclew hnalysis temperature I _ tee _cw Ne gee Next gt Bidcore S51 Instrument Handbook 42 3
115. oechoh points 8 F me ropetes OO ceek Hens ewe 51 Flow rate 90 l min Number of cycles 5 Run correction Cycle every__sample cycles 2 Compound Characterization 1 Next gt Biacore S51 Instrument Handbook 3 91 Compound Characterization Injection Parameters S ampke nechon Contact kre Dissociahon mmer ha FF Exra weash ater iniectior wmhr 50 DMSO Stabdizabhon pesiod talEatiC kre Help Next gt iliE sample Table SSmgis Pasitan mw ajjcncJaw eonc Tom 1 Fuosemide nD Furnsamide R12 BdEg 3 3 IFurnsamidm 1L3 Ba 3 Furnsmmid PT by HHa 3 3 Furosemide R1a5 1 35 3 75a 6 Furnsemidg B27 5e 3 Furnsemid FR 1 E55 Furnsami ui Eg Furnsemid PH BEg 10 Furnsami
116. ots harkers Statistics Harkings Characterzation Markgr Lnulumms fBaselinc Earrprnyer Bindinn to reference spot Controls Binding lewels Controls Bindinn stability Cancel Statistics Tables Calumns eneral Fl Markars Slashics Hankinas Characterization Lnntrn 5 ample Statishics Lnlumns c nl r corr OD Controls Binding lewels SD Fc 1 5hnl 1 T corr Corntrols Binding lewels 2 Fc 1 5nnl T En Controls Bindinn lewels amp wg Fc 1 Spot 2 r corr Cortrols Bindinn lewels SD Fc Spot 2 r Cor Controls Binding lewels 2 Fc 1 Spot 2 r corr Ol Cortrols Binding stability ng Fc 1 5nnl Controls Binding stability SD Fc 1 Spot 1 Ol Controls Bindinn stability Fc Spot 1 Controls Binding stability ng Fc 1 Spot 2 Ol Controls Bindinn stability 5D Fc Spot 2 Controls Binding stability Fc Spot 2 Corntrols Bindinn stability amp wg Fc 1 Spot 1 r Controls Binding stability Sh Fc 1 Spot 1 CO Controls Bindinn stability Fc 1 Spot 1 r Controls Binding stability amp g Fc Spot 2 arce Biacore S51 Instrument Handbook 78 3 OK Cwcle Cwcle Type IFPnsrtinn Samples Hame pm 1 Fc 1 Spot 1 F corr 1 tatup uffgF 19 4 a tartuph Huffer 4 tatup 1 Buffer 3H 4 tartuph 1 Huffer
117. pHscouhng 140 0 318388 385 No 2678 3 10 mhd_Acetate pH 550 10 20 Carbonic anhydrase 1 1 pHscouhng 191 0 2942E1 089 Yes 255 5 baselne2 10 20 Carbonic anhydrasa 1 1 pHscouhng 311 0 374465 15 92 No 8020 4 10 mhd_Acetate pH 500 10 20 Carbonic anhydrasa 1 1 pHscouhng 3520 294833 0 79 Yes 53 2 baseline3 10 20 Carbonic anhydrase 1 1 pHscouhng 4820 402765 18 18 No 10787 2 10 mM_ Acetate_pH 450 10 20 Carbonic anhydrasa 1 1 pHscouhng 5330 29671 9 1 27 Yes 182 6 baselined 10 20 Carbonic arhydrase Event log Sensorgram Fc 1 Spot 1 Time Imio Time 731 0s Response 29372 5 RU 00 Tmestamp 28 Mar 013 13 30 20 00 SelTemperalule 25 0 TT Flow cel 1 00 Tempeialure 25 00 CT 00 Sample Temp Seting 25 Temperature 25 00 C 00 Fiowcell1 Standby flow max remaining time 4 00 Days 00 Flow 30 yimin Immobilization pH Scouting Standby flow Standby flow Biacore S51 Instrument Handbook 2 25 Immaobilization pH Scouting Results E RU 14000 PH 4 5 13IHH 4 TOOn FO bf Bd EOD HH WEFIILUIT inn 3D Eh 1 120 147 IE Timis Ligarmd Curve Bufgr Fal esp EI ET A 10 mid cetae pH SD eB
118. qbility lote co _staqbility_ late 2 Use two solutions Address spots separately Stabilizqation period Adaptive 30 RU Carry over control Adaptive binding_lqte stability_early co_binding_late co stability_early Biacore S51 Instrument Handbook 60 3 Hit 5electinn Imjectin ELuEhcE Capture E
119. rH FIaw tel 1 nh rg BU EE 10 mh Acetate pH 150 10787 2 nn 1 mi ncelale ME Hg5H g Rel Resp pH pH 10ug mlin 10mM Acetate pH5 0 gt 100 ug mlin 10mM Acetate pH5 0 pH4 Biacore S51 Instrument Handbook 26 2 2 2 Immobilization Menubar Run Run Wizard Tool bor A 2 FT um Wirard uface Preparation Innhll ah Immnbilgalinn nH Scnuling
120. rcleam Test Taals ll Noemalige El Sustem Check HO Sevice Tools This procedure remowes adsorbed matenial ared dsintects fhe Hr EE EE hn rint run this nincedure Bel UL TE se IhE ainterarnce Chip for Ihis necedure he sumlace mm other sETSOT CFDS may be Harrsned by he sluliOri usesd Last Hun Tme UEApr03 21 00 Help Farfnrm tool king Larr BIA BIAdesorb solution 1 0 5 SDS 4C BIAdesorb solution 2 50 mM Gly NaqOH pH 9 5 Desorb Start This nrnceHurE remowes adsorbed material Irnm the nw gmstem Total rn kine 1s abonut 22 mites NOUTE se the hairntehnance Chip For this nrncedure The surface nh other sehsor chips an be damaged hn the solukiors us hin ot ruh this hrnceHurg belo CE ark Biacore S51 Instrument Handbook 4 115 Next gt Desorb x Fiequired solutions frorm hd airnternance
121. sesesessssssssssesesssseesesesessasssessssssssesssessseg 1 J 10 Zoom Lock Decisian Heject rate constants Help Cancel MTL Biacore S51 Instrument Handbook 3 3 3 4 _ Menu bar Evaluation Compound Characterizations EL Campournd Characterizations Lnmnnurid Characterizatiorns Lfmide Fc 1 Spot 1 F corr es dTarnIl 103 CHS Fc 1 SFint 1 F corr Yes Furosemide Fc 1 Sphot 1 F corr Yes SulfanIlaITiidHE FC 1 hnt 1 F EnFF es Delete wt Deactivate Llnsg Add Edit
122. spot 1 lsT Fusinn nrntein 120 Lnnlact hm 1 Flow rate 1 11 mnl Exha wash after inection 1 Linand spot lsT fusion proteirng 1 Fleuwrates 10 Lnnlacl hm g mnl Exzha wash after inlclinh 2 Fanlure stabilizatior hime Sample InIEChinhn Contact hime lg hisnciatinn mg 3D wv Ezha wash after inection with hi Duplicate samples Hgneheralinhn Hgneheralinh snlulin EE pHEHnHgI Lnnlact mg High yiscnsihr snluHinm Stabilization herlnd Gtabilization lime s He Cee 2 1 Biacore S51 Instrument Handbook 3 59 Adaptive Hit Selection RU RU binding ate lola 30sec 1 SHE TI Carry over control abilization Redeneration Regeneration Adaptive sta
123. t Selection lt 48 3 2 1 48 SID EF ME hc ii 61 3 2 3 XX XXL 64 3 2 4 Sain 81 3 3 Compound Characterization ii 89 53 oo A 89 2 93 3 3 3 Sk 100 958950 1 103 3 4 1 bound es 107 3 4 2 9 bound 4 108 4 SS 113 EL i 44 114 4 2 tt 119 5 122 351 122 5 2 kk 124 Biacore S51 Instrument Handbook 1 1 1 1 1 1 1 1 gt gt Ready
124. ty of binding Use this wizard for etarmple on lead optemizatiorn and MSA wok Hep se Tempnlsle Compound Characterization Start Eo ee 1 2 tre sabilization perind hhPLE 7 Sample Beqeneration sg tn saitions es st 1 WAIT ly 5tabillzatian perind HHT VER Larrp nyer cnnrnl He rk Llnsa Biacore S51 Instrument Handbook 90 3 49 Hit Selection Next gt Compound Characterization hy etup _ 3 Deateriion Flow cael Spot 1 gE Feteterces PH Huim TUF caw ralm Flowy rabe 9m mnl nndihoriri Fu sernsor chip cnrmdihtorr Ile wm HI Fr Ru startup cycies S armele SOHO Buffes Numbm ol ccjer Snhyent crechnm Fr Fiun snlyer conection CUCles HUm cothecthon cci ey lz ample COTIETWTII fm sees Nummbes oF c
125. u Select all 25 Lucle 1UB arhasubenzene sulfnnamide 125phd Cucle 1UE rhnsnhenzere sullnnamids b 25p hl Deselect Al Cucle 107 4 Carbonnbenzene sulfonamides 12 hp 15 Lwcle 108 arhnsubenzene sulfnnamide 12 5p hi pS Cucle 104 4 Carbonnbenzene sulfornamide Sp hd 4 cle 11 arhnnuhenzene sulfnmamide 5 0 5 0 2e 5 4e 5 Be 5 ge 5 1 Con Kinetics 1 1 binding model kg kg Rmax 3 Constant Local Global Constanl Local RI k kK Rmax Global Rmax Global
126. uality control plots Baseline Carry over Bo Bindina to reference spot i FT Controls Binding levels wn a aki Fadi She Eg NY Fesult plots i Bindin levels 1 ES Bindina lewels yersus stability Binding stability early RW PF stabiiy tate a eSult Table Sensorqram Displas sensararams for analysis cpcles either singly nr in oyerlau plots Cycles hat are excluded from the evaluation are shown with a broken line in the sensorgram 12 Helerence Fc 1 Spot r Biqcore S51 Instrument Handbook 3 65 Binding levels Evaluation Explorer Binding levels binding_late Binding levels 140 120 100 gt 16 Azosulfamide 80 60 40 Response binding_late 8 Control Furosemide gt 15 Furosemide 18 Control Furosemide gt 13 Sulfanilamide 14 CBSA 20 7 Negative control gt 10 Warfarin gt 11 Napoxen gt 12 Phenytoin 17 Negative control 1 Buffer 2 Buffer 3 Buffer gt 4 Buffer gt 5 Buffer gt 9 Digitoxin 0 20 0 2 4 6 8 10 12 14 16 18 20 Cycle num ber Buffer Negative control
127. yent correcton Lanc Table Columns General Plots Biacore S51 Instrument Handbook Table Cnlumms General Flots Markers tahshics Hankinas Characterization Flot Columns Bindngleyes Eindinn leyes Bindinn stability early CE Binding stability early CE Binding stability early El Birnding stability early indinn stability ate Bindinn stability ae Bindinn stability late Bindinn stability late Baseline Baseline Baseline Earrp nwgr Earrp nwgr Bindinn to reference spot Fc 1 Spat 1Tcarr Fc Spot 2 r En Fc 5nn Fc Spot 2 Fc 5nnl Fc 5hnl T Fc 5nn Fc Spot 2 Fc 5nnl Fc 5hnl g T Fc 5nn Fc Spot r Fc Spot 2 Fc 5hn T cnrr Fc Spot 2 r En Fc Spot r Lancl 3 77 Binding levels select All Deselect All Markers Table Calumms General Pl
Download Pdf Manuals
Related Search