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Biacore A100 日本語取扱説明書

Contents

1. OANA HE SP lt e5X33J2 lE Pe 22202
2. DMSO Biacore A100 126 6 Biacore A100 DMSO DMSO
3. FEADER J DACIA BRR EBBJ LERE CIA 6678051 BEBEDRISU 7 873 py STE fe RE XEZI S AER C ep UL COSS 9 c clc UL SL ng m Ze ug ml Ko B 1 10 10
4. OFF TEL 03 5331 9336 9 00 17 30 WwWw gelifesciences co jp e maill Web 02009 GE
5. pH 25 1 2 pH RI pH Scouting pH 150 mM NaCl HBS 50 mM NaOH
6. Biacore A100 66 4 4 3 4 3 1 Regeneration Scouting 1 New Method Biqcore Methods Assay Development Assay Conditions Surface activity test OK imi yupin Ligand activity best Bit baigi bhira jur 25 7 sange Sample Fire 2 Seling lor step Ligand scii test 1 CT T suns 25 Micah m ufo hende Nu Hn 5 i a Sample 1 120 s 120 5 Opinias pr Horat DENT Sample amp Assey Setup Al General Settings
7. Response Time Biacore A100 56 4 JH amp R TEFHRIGE OD TS 600D2E P H8 HEIS ES pH Og NaCl 2M BEER E 10 mM Gly HCI pH 1 5 HCI 100 mM Phosphoric acid 100 mM Formic acid lt 20 10 mM Gly NaOH pH 12 NaOH 100 mM Ethanolamine 100 mM Ethanolamine HCI 1M 210722 4 EDTA lt 0 35 M FRESIEK Surfactant P 20 Tween 20 lt 5 Triton X 100 lt gt SDS lt 0 5 96 Octylglucoside 40 mM Acetonitrile 2096 DMSO 5096 Ethyleneglycol in HBS buffer 5096 Ethanol 2096 Formamide 4096 Guanidine HCI 5M Urea lt 8M Biacore A100 4 57 4 2 1
8. RU x S Rmax RU Da Da S RIJ 50 000 1 000 RU S 1 20 000 Rmax Rmax 20 000 1 000 x 1 50 000 400 RU Rmax 50 100RU Rmax x 5 Biacore A100 26 3 3 2 pH
9. 3 GE T 169 0073 3 25 1 TEL 03 5331 9336 FAX 03 5331 9370 e mail Tech JPG ge com ISO 9001 2000
10. Ks Surface Activity Adj 0 100 0RU 100 Signal Correction Setup P Surface Achwity Adj P Results Sunlar Aci Adpiriment be oi repoitpani n zelecied cuves aERERERERRERERERERERREREREREREREEREERER m Balch xeecl curvelupe al Balch repoitpani far al raz bindra las Control Linear E EE eph TT eene T B 3 a betazmiero 5 un mi TTE Pos control Fcl Linear 0836
11. DMSO Biacore A100 132 6 6 3 2 Screening Kinetics affinity 1 Create assay steps by et h O using the buttons below Startup Startup Sample 3 times as entered 2 Select an assay step by clicking on it Edit properties 1 Overview below or use buttons to Sample remove or move the step Sample Sample 1 time as entered 4H K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K K NN FPF General Settings New Step Solvent correction a Solvent Correction Sol corr 1 time as entered Before every 50 cycles Fiemowe Step Control sample Assay Steps __ MeeUp t Control Sample Sample 1 time as entered Before every 20 cycles Move Down __ Mise Hee Semple amp Assay IL Setup Assay Steps Solvent correction Recurrence Recurrence Repeat assay step
12. 90 EE N Aun g 3 D CM5 NHs N o Up Ege um NHS COC CONHCH CH OH lt gt Series S Sensor Chip CM5 BR 1006 68 Amine Coupling Kit type2 BR 1006 33 EDC NHS 10 ml 200u 22 UL C 20C 9 S 1 4C 59 5 BIAdesorb solution1 2 BIAmaintenance Kit NaOH 50 50 mM NaOH BR 1003 58
13. E RU Adjusted sensorgram G 6000 E pH4 0 5000 2 2 pH4 5 4000 pH5 0 so 5 2000 pH5 5 8 1000 Ai v e 0 7 1000 UmM 3000 50 0 50 100 150 200 250 300 0 Sample 1 Start Biacore A100 3 27 3 2 1 pH New Method Surface Preparation Z pH Scouting OK Method z Drimis kr Low Sanpla G T SI mH ED s Method Buffer Fc1 HBS EP IHBS EP Buffer Fc2 Buffer Fc3 IHBS EP Buffer Fc4 HBS EP Solution used For running buffer in Flow cell 4 m a 4 5 Concentration unit ug ml Y Biacore A100 28 3 Sample amp Assay Setup
14. Exclude Curve Assay Finnl Beta Chwa 72 3 Kinetik betazmicro 20601 5 0505905 chwa Fk Vaw Hob Tharbevis sy Cue z Scde 77 Descriptor Based QC S 2 Sova Continue _ se sw EE We ee 11 Veit Biacore A100 184 9 QC Summary All sensnrnrams J Controls sensorgram e Samples sensorqram K Summa D ER WANNERENKA Barre A100 Evaluation Final Hz Foi r hia T 3 5 Kinetik beeazmicro 20811 5 mes chwa Approve Data zelhimqs 219 i L i I qi m mi LIS E Eretee ote ee eintesacond iae seess Biacore A100 16 Approved Data Microsoft Excel File Edit View Help Close Save Data Save P
15. ReqJ 8 OTA Neq Ko Rmax Rmax 2 Biacore A100 Kp C MJ 2387274 HI Ko 1 10 10 1502 1 9 5 EE 5 0 1 2 n 2 10 0
16. gt X x Export Table Save Save As Save Finish 221 22 100 17 Ranking binding levels Evaluate Data Settings 49 Plot 49 Result cave Scu FE1 1 3 61 acm acr Fet T d pgri FEST a ibai HSA ipai LA Au zj f Ledmet 1 7 Uniockedpkt Back gt Exit Method l
17. 3 ImrrnrE A100 Evaluation Arsay Kumagai basic datasza0n5 Kinetilk a40525 1 202042 Edt Hp Evaluate Data Settings 4b Preparation b Examination Results Table Azosulfamide Salbe kd 269620178 _ i ed 1 042 i KD 273207 M 0 3197 5 i Wa lion Sersorgiam Table Parameter Conc Ini Values Resdua M i jon messege ls E 5 3 H 3 1 3 22 3 pon Caborer ram EE 3 Apaadi TU Exit Method Evaluate Data Affinity Concentration kinetics ResultPlot Sensargr am New Kinetics Affinity 9 100 218 9 Evaluation Items Kinetics2 Affinity2 sea 7 Kinelics Evaluate Data
18. Include oetup Select corrections to apply Solvent Correction Solvent Correction Additional dialogs will be shown Molecular Weight Adj Molecular weight adjustment compensates for the fact that a large molecule gives a higher response than small one by dividing the response by the molecular weight Capture Adi Capture Adjustment The run does not contain any capture injections Data cannot be adjusted Surface Activity Adj Surface Activity Adjustment This correction should be used only in i screening assays Biacore A100 Solvent Correction Solvent Correction Next gt Signal Correction Seiup SohentComechon P Molecular weight d p Acimty k Results Cobani Cac gas Du Direction al Cbs L Fa 5 Debe
19. Kop M Ko A B AB Kp A B AB y r F ks Miis A B ka Ka D kao ka BREES Ko 1 1 1 binding Biacore A100 200 9 R f ka Rmax ka kg TA 7292 Baat p lol 4
20. CIR Biacore A100 1 Biacore 100 5 077 7 E37 V F UI sil r9 55 Reagent Plate BR 1006 08 Microplate gt e Reagent Plate and Fol 24 1006 08 Microplate 96 well 96 BR 1005 03 Microplate Foil 96 well 96 9X BR 1005 78 Microplate 384 well 384 BR 1005 05 e Microplate Foil 384 well 384 7 BR 1005 77 100 6 2 2 2 1 2 1 1
21. 0 Comment Control curve below x axis PERAN Biacore A100 E1595 Quality Control Approve Data 2 gt v Z0 Quality Control i Harare ALOI Evaluation Assaw Final Bela chia 7 3 5 Einetik bebazmacrn A0015 D 55585 chua c p e Fia Edt Descriptor Based Quality Control Approve Data Basalina ualty 4 Association Quality 4 Dissociation ul 4 Summary Tabla Association Outlers i 19 Fc3 2 3 a beta2miera 5 ugimi 9nM 54 Descrip Based GL S Pix And Continue ee mW QC 3 A Descriptor bqsed B Plot based QC C Sensorgram based Biacore A100 180 9 Descriptor based
22. EJ EIJ E Biacore A100 20 2 826 E Toolbar PaK tia nsert Eject rack trays 3 x Please insert Z ejectihe desired rack trays The hotel hatch will be closed automatically vathin one minute if no action is done Positiones Empty Reagent plte 384 wall Microplate plate 384 wel Microplate plate 384 well Micraplate Empty Rsagart plate well Microaplate Emptu 24 22 E 21 202 Fieagent platz 384 wel Microplate Fieagent plate 384 wel Microplate 21260485111 16 18 pate 14 OCC 384 Wel Microplate HH H HHEH 11 10 Reagent plate 8 OOOO LOLL 384 well Microplate H H E H HEHH B 3 2 OOL 1 OOC H Help Ejec Tray Eject Tray Insert Tray
23. gt VT mU bxc 96 2 lt gt Startup 2 T 3 Sample 2 10 mM NaOH 30 Control sample 20 1 ae pe gt 2min AREE 2min bra 30sec Biacore A100 1044 6 XV v gt New Method Biacore Methods Screening Screening OK Sek imis Mam sie wai lon GU S General Setting 96 Assay Steps Biacore A100 6 105 Assay steps Method Control sample ae Ltmeasentered Before every 20 cydes 1 Assay step
24. BE Bed CU Ss 2 Biacore dor c 6 2 Sensor Chip es CM5 oaBeries Biacore A100 2 1 BiacoreA100 v Fc 4
25. Keyword Table Report Points Keyword Table selling W Tha wakapata kd iha kavandi cures in iha selected dala Emad shown on s sinite bsa cnn or canit Em changed Lise fnis In eraus het sd ane eniered proceecing eih fe dels approva Dick an s column header bo cort the tbis by the potens pf Is column 1 alal asee smn son 13 4 50 1 di 3 si CA pH 1 3 Sanple 30 pugil HA pi 4 5 SO mf 2 2 sp 45 RENE 2 3 3 smpb j amp OugmiCApHAs Bonm NaOH J 4 30 4 5 NaOH 3 1 3 Sampla HA pi 5 Hace 32 35a 11 Ba pet SO mi NaOH 3 3 3 5mpb j amp pam CApHS SOmMNSOH 3 4 xu CApHS 0 SO mM NaOH 41 35amph apam S S NaOH 2 15 5 5 SO nM NaOH 3 33qmmm CmuamCmpHS5 SOn NaOH 5 5 50n ANd 100 174 9 14 4
26. Hecurrence vw Repeat assay step within Sample Every po i cycle Distribute evenly Run assay step once first Run assay step once last Repeat assay step within Recurrence Every Distribute Run assay step once first Run assay step once last Biacore A100 5 83 Assay step preparations Assay step preparations Temperature 25 Normalize after temperature change Refresh 25 O Menu gt Set Analysis Temperature Normalize after temperature change Refresh Biacore A100 8 5 4 Cycle Types lt
27. gt ug m E E Biacore A100 4 47 4 1 4 1 1 New Method Biacore Methods Assay Development Assay Conditions Binding conditions OK urcerdtalion unit Lone d ue alie 28 7E Control tange 1 hirre ars control Corro Sample Sange 1 hive eted tte igaerr 3 odes Pq Sample 1 BD s Ef x ko Horal 5 Reprisal T walmt by cell 30 z Cam ove 1 Method Description Buffer 85 2 5 used for running buffer in F
28. Apply same spot settings to same spot in all flow cells Apply same spot settings for all spots in a flow cell Next gt Biacore A100 9 167 Approve Data ILJr gt q Li 1 CO ED 2 atr EL BL 4 Feti 9 Fz 3 m at 4 r 1 T 1 m RB 1 T r AM nr 1 mun i E BHE se Include Curve Fc Spot Ligand Spot Setting Reference Subtracted Reference SAS ue Active ekdeAl inedeSaeoed EreudeAl Tables Biacore A1
29. Descriptor Based E Baseline ml Larry over tj Binding to reference spots ij Controls binding levels 1 1 Controls binding stabilite Sensargrams All amsnrnrams Controls sensargram 4 Samples sensargram Summary View Sensorgram Exclude Curve Settings 4 Plot 4 Result Thumbeols Sart ba Curve y Ed sede Biacore A100 Sensorgram based Descriptor Based Plots Baseline gt Carn aver 1 Binding to reference spots kl Controls binding levels Controls binding stability All sensorgrams Controls sensorgram Samples sensorqram a
30. End Row 1 StartRow End Row 96 1 12 Stgrt Row 2 End Row 1 1 2 A Menu WV 7 5 Use bottle for reagents Yes Biacore A100 102 6 6 6 1 3 RU ORI A or wo D Startup gt IRIS mm e Xu tL U meuzri5sg 2 Solvent correction 124 6 3 3 Control sample 2 l EJ am 2 Sample Biacore A100 6 103
31. o Wax Cycle Run List z und Biacore A100 5 75 5 1 Action 77 5 New Method i E new et Methods a pH Scouting Assay Development Pe Assay Conditions Regeneration Scouting Screening Kinetics and affinity Concentration New ee a Surface Preparation Assay Development Screening Ds Pu Kinetics and affinity Concentration New OK Biacore A100 76 5 Sack Y 121 gt Lo Consurgicn 1 50 mj
32. Biacore A100 100 5 5 2 CO SO a BD Pooling Tray 113 1 1 Try 82 2 Tray t3 Bottes Reagent plate Pos control Fc1 Pos control Fc2 uM Pos control Fc3 pf Pos control Fc4 pM 66 Pos corkrol Fc1 uM iPos control Fc2 control Fc3 control Fc4 pM 66 Pos control Fcl Pos control Fc2 uM Pos conrol Fc3 Pos control Fc pM 66 Pos control Feci Pos control Fc2 uM Pos control Fc3 pf Pos control Fc4 66 Pos control Fc1 uM Pos control Fc2 control Fc3 Pos control Fc4 pM 66 Pos control Fc1 Pos control Fc2 uM Pos
33. Validation infa Sensorgram T able Sensorgrams Parameters p Initial Values usen tuve cone conc unt CA 2Dug ml in 5 1 1 3 E PE 22 2 e i NK M bo Am a DNE 5 PTS PES EE e T a EE Uem I E A20ugfl in pHs Feli 2 c A MC VM D am oo m N me iw NN Biacore A100 Sensorgrams Validation info Sensorgram Table Sensorgrams Parameters Conc Initial Values Furosemide Response 50 50 100 150 200 250 Parameters Validation info Sensum Lai ME Parameters Conc D Initial 0 9 9 Validation infa 5Sensorgram Table Sensorgrams Parameters Conc 0 Initial Values Biacore A100 214 9 Initial Values aldalinh info Sensargram Table Sensorgrams Parameters Cone Initial Values Initial values 1 000 5 H Next gt Y X The
34. Toolbar EMEA 7 77d 2 x Rack tray temperature 81 Cancel 4C 40C OK 25 Biacore A100 9 10 2 2 2 2 3 55 l i Biacore A100 2 11 Fis Tons rabrurenk a qu Chp Cholomale Temp Hack Tia Tarep Chip and method information Chap kot nursber Chip ChipID Metwdi Setup Run he Homse Thi procedure eimaizes the deiu response al Es cell lima about 10 minuse Da not use fs M enbensnce Chip F Addeessing piaceduie callialer Em ides cared slderzng ol deleclion spoit vallin e celi Run brse 24 rn
35. thon unit E x 2 US GE SES 4 D F DI F D F P IP PHP P P sce oT Les DP Dispensing pump A B 1 2 3 4 4 4 96 384 8 CQOOOOOQOCO 2 FLLLLLLLLLLLLLLI 000000 0 9 KILELENI ee O O eo eo ee J J Biacore A100 4 1 Biacore A100 4 1 Til LA 2
36. Solution T Contact Time m n Width spots 40 mM E mH 5 7 15 1 2 z ugend ___ Solutions defined in Setup Form l esctvstion 1M Echenclamine 15 1 2 iss Mines gt Hove Down Lancia Purpose 7 9 707 7 SXNIUBBJeEIRS 5 Activation LicancdDilute Activation EDC NHS CrossLinking Deactivation Reagent Ligand LigandDilute CES Biacore A100 3 45 Width Width spots 12 B 1 RIY F 1 3 1 4 5 5 1 2 1 3 1 2 4 5 4 5 1 3 1 3 1 2 3 4 5 Add
37. DMSO 1 d DMSO DMSO 0 M SIRS 4 DMSO DMSO DMSO 9 100 DMSO 0 4ml 10ml 6 DMSO nc EN DMSO 9 4ml B A AR EAR 100 DMSO 0 6ml 10ml 0 0 SER MIS FRU l p 100 DMSO 50mL 4 69 X8 25 925 Y 8 4 DMSO 200 400 600 800 1000 1200 1400 ul 6 DMSO 1400 1200 1000 800 600 400 200 SAONA 10mM in 100 DMSO j x20 DMSO 500uM 2222 7 5 DMSO x50 188 5 DMSO 10uM in 5 DMSO 100 DMSO DMSO J l2 C04 8 DMSO DMSO DMSO
38. Solvent Correction Remove Step CHIERS lt 2 Base settings 228 3 1 3 Control sample Recurrence Repeat assay step within Sample Every 20cycle Run assay step once first Run assay step once last Biacore A100 106 6 2 Number of replicates Number af replicates fn 4 times As entered 1 2 3 1 2 1 1 1 2 2 3 2 Random Startup 3times As entered 1 2 3 1 2 3 5 Assay step preparations 25C Cycle types 100 6 107 Cycle types mm Toas ie EL m Chip Hmpimcmehan Tena Flack Tray Temp F sck Tray Heels 1 CrEalE a new cycle type or select exeting irm de lisi Method Thus is ugad md conii sampa ri x 3 Configure Por auch comman CD Cycle types currently in method Sample
39. Sample Commands Sample Insert Command Remove From Cycle Move Move Down z Regeneration 1 i Cary over Type 2 20 F Low Sample Consumption Injection volume 25ul High Performance Kinetics niection volume 50ul Normal 2 Injection volume 36yl Flow rate 10 30ul min Contact time Dissociation time Type Morral m Flow rate 30 El min Contact time lgn s Dissociation time 60 s Biacore A100 88 5 Regeneration Commands Capture Regeneration Regeneration solutions Regeneration F Flow cell 1 Regeneration Fc2 Flow cell 2 Regeneration F3 7 Flow cell 3 Regeneration Fet Flow cell 4 Contact time 30 s Insert Command Remove From Cycle Move Move Down Atleast one high viscosity solution Regeneration solutions 25 2D
40. Assay Steps Biacore A100 Method Poeni General Settings Assay Sieps Cyce Types l E aamgle Asse Setup Actions L jnes 4 67 Empe Sampe 100 times 35 enibenad Fecurrence Repeat step eilhis Every re F Distribute event Ame T Fon aga pog el preparation u Tarspearstura E femeni HEATER M T7 Ne aiar change m Order 12233 m EN Number of replicates 100 2 Copy Step 2 21 v 23 Activity test Sample 1 time as entered A Sample of Activity tes Sample Sample 1 time as entered Activity test Base setting Name amp Activity test2 JJL Number of replicates 100 2 200 Biacore A100 68 4 JHERTEFHRIGE OD IS 600D2E T HIS Cycle Types Cape H
41. Setup Run Sample amp Assay Setup Biacore A100 92 5 Verification utku o Method has been verified be used for running methods on the instrument C j eR Save Method As _ Tee Immgkiisaon Method 12 9 2004 1 31PM FrancisMarkey 1 12 7 2004 11 3 77 Metoddogi S UM os 88 5 71 Software ta Inmoiizabon Method 9182004 8 35 AM Method Buider Method 12 8 2004 10 24 Y Method Buider Method 12 23 2004 2 03 PM Francis Markey Immobiliza amp on Method 10 1 2004 12 47 rancis Mar i Descrpton Help Biacore A100 5 93 Run
42. Baseline Quality Association Quality Dissociation Quality xE 88 B8 amp 3E Bx 43 43 4t E LAS fest Se DS HA 5 SAARTS F 5811 2 00 861060 2765 1025 1 1 SE Harare LED Evaluation Aesaw Final Betas c haga 73 5 Eimetik hetazmarcrn 24811 5 DIS chua Descriptor Based Quality Control Baseline Gualty 4 Association Quality 4 amp Dissociation Rualry 4 Summary Tabla Association Outliers 819 3 2 3 a betazmicro 5 ugimi bam 8nM e lt gt sc mem iO pu cope Exclude 2 Next gt Baseline Quality Association Quality Dissociation Quality 08 Biacore A100 Summary Table Descriptor Based Quality Control Baseline ualty 4 Associati
43. Biacore A100 3 41 7 UOS VIEWS GST FLAG protein protein G 2 Cuv x 5 2 4 PRIJA PEE 5 uir gt Immobilize for capture Flow cell 1 Chemistry Spot Usage Ligand Contact time mirn Use spot 1 3 10 min activation v Lise spot 4 5 Amine coupling 10 min activation v Imrabilize For capture 1 5 2 4 Biacore A100
44. TREA 4 Biacore 05 27 227 7 12 x10 HBS EP 10X 1000 ml BR 1006 69 0 1 HEPES 1 5 NaCl 30 mM EDTA 0 5 96 v v Surfactant P 20 C 10 f amp 538 0 01 HEPES 0 15 M NaCl 3 mM EDTA 0 05 96 Surfactant P 20 pH 7 4 HBS P 10X 1000 ml BR 1006 71 0 1 HEPES 1 5 M NaCl 0 5 96 v v Surfactant P 20 C 10 18 7578 0 01 M HEPES 0 15 M NacCI 0 05 96 Surfactant P 20 pH 7 4 HBS N 10X v 2 1000 ml BR 1006 70 0 1 M HEPES 1 5 M NaCl 10 5253 0 01 HEPES 0 15 M NaCl pH 7 4 PBS 10X 1000 ml BR 1006 72 0 1 M phosphate buffer 27 mM KCI 1 57 M 10 5784 0 01 M phosphate buffer 2 7 mM KCI 0 137 M NaCl DMSO 596lv v S pH 7 4 0 22 IN cce os Biacore A100 8 2 2 1 3 Start BIA programs gt A100 Control Software 0527425 Biacore A100 Control Software Username admin Fassword
45. Ranking binding levels 1 sos 4 Plot 4b Result T abis Sot bp cave a FE1 1 3 eon Fet 2 3 eon Fei d cor pgi Fez 3 eon H L And Finish Fez5 3 car AGF 1 3 680 HSA 2 Exi Mead x u PEE Mises i Mena avara 5 Pn f Uninckedpiot Back gt Exit Method Evaluate Data Affinity Concentration kinetics ResultFlot Sensorgram Sensorgram Biacore A100 9 195 Evaluation Items 2 Sensorgram1 Evaluate Data w ana Argi Finish F pprosed Dala Hem E LJ liina 11 Ranking bindrgle p Ranking bindeg H Bindrg es 34 KK KK K K K K K K K K K K K K K S KASS S S S SSE S K K EI Sensorgram 1 Setlings d Senserqram m PSD B In
46. v Setup Run Ligand overview Spot Flow cell 1 Flow cell 2 Flow cell 3 Flow cell 4 GST proteini 2 Act deact 3 Unmodified 4 5 Run Order Chemistry Amine coupling 10 min activation Flow cells 1 5 pots 1 3 Ligand injection Contact time min Spot Flow cell 1 Flow cell 2 Flow cell 3 Flow cell 4 1 15 1 GST protein buffer buffer buffer Positioning B Trays Run Biacore A100 3 39 3 3 1 2 2 1 1 2 Flow cell 1 Chemistry Spot Usage Ligand Contact time imini M Use spot 1 3 coupling 10 min activati Imrmaoubilizec nti 10 Immaobilize for captur Immobiize T anti Igiza 10 Overview i Chemistry Amine coupling 10 min activation Flow cells 1 5 pots 1 3 i Ligand injection Contact time min Spot Flo
47. 3 Edad Land nasa Ehdsul Selected Inmetate Sabasa Escas Selected Make following cycles available Make following curves available 198 9 tf gt Sensargrar Thonsbesis Hew Fites Tools Zombe Sen by Cave Dem Sce Adjusted gensorgram m J 100 2180 apa ax 560 eno Time 2 Sample 1 Start mm mu Ctl Sensorgram1 Setings 4 Sensorgram Thonibessis Tools Fis Adjusted gengsorgram 9 2 2 D water
48. Method Doral patom immobilia on ar temperatures Ier thar d 1 are thal homamatior and Hydindynamic Adde sirg haue baen nan prior ip immoblizalinn 2 Add igand and rescents In Ihe specified postions 5 plab with Foil 4 Placa on lhe ard Iha insltumeri 5 Fil baila B il immobiliestion bulle and botas 1 4 vath buffer Lor ber the immazgiioni run has finkhed amp uamalic sich urs a n ange nl bufer ill blake sine Ihat the ubings reach the bokom cf iha E Empty thie washa can T Stat run CD Chip type CM5 SA NTA L1 Custom 2 Same Settings for all flow cells Biacore A100 3 37 3 Chemistry 5 10 3 Flow call 1 Chemistry Spot Use spot 1 3 1 Amine coupling 5 min activation Amine coupling 10 activation aurface thiol caupli iw Use spot 4 5 5 2 Spot Usage Ligand Contact time amp U F S112 Usage
49. 6 Solvent Correction Sample 7 Startup DO HAE Connect to cycle type Cycle Types 6 gt Startup lt Name Startup Purpose Sample 3 times as entered Connect to cycle type Number of replicates Biacore A100 82 5 C Number of replicates Number af replicates fn times As entered 1 2 3 1 2 1 Order 1 1 2 2 3 3 C Random As Entered Order Random D Recurrence Control sample Solvent correction Calibration Sample Sample Sample Sample 1 time as entered an Control sample D E Control Sample Sample 1 time as entered Before f every 20 cycles
50. DMSO Biacore A100 128 6 6 3 1 DMSO DMSO 2 DMSO Action New Method New New Method Builder method OK E New Method X Biacore Methods ace Preparation Og Immobilization pH Scouting m Assay Development Assay Conditions New Method m Regeneration Scouting Builder method Screening 751 Kinetics and affinity 7 3 Concentration New Use this ta create a new Method Builder method Cycle Types Biacore A100 6 129 Method O Cycle_1 DMSO check
51. 5 New Method Biacore Methods Assay Development Regeneration Scouting Regeneration scouting OK EN UTE Lnrrerltalgm uri uhi 5arnple Sampi G tines 3s entered A Eeg conditinn 2 Sampi es entered Selbnos lor step Fleg condition 17 4 Heg canditinn J Sample Sample rires ss entered A 1 i i 1 Sample 1 0 s ur Love Sample Coram 4 Fanperaealinn 1 Sehr vare 30 General Settings q 2395504 RESM 71 Assay Steps Biacore A100 58 4 1 Create assay steps by using the buttons below Reg condition 1 Sample Sample 5 times as entered 2 Select an assay step bu clicking nh it Edit properties below or use buttons to Reg condition 2 remove or move the step Sample Sample 5 times as entered A Reg condition 3 Sample Sample 5 times as entered Remove Step Reg condition 4 _ Sample Sample Mowe Down S
52. EIL OL XE Or OC YC MC 192222222 1010000000 100000000 M 22222222 i 1000900060 x XL C COL C XC X OOOOOOQQO sQOOOOOOO 000000060 1 OQOOOOOOQO 29009900 e gt Bottles S 222 V u 296 SESS TEIG TS Estimate run duration Tray 1 Bottles agg Estimated run duration Position Content Estimated consumption Water 100 ml ECHTE bottle dead volume and standby 390 W ater Biacore A100 98 5 rrays setup Insert Trays Biacore A100 5 99 9 Start Run Antibody 71 Concentration ey demo DNA DNA liposome 1 manual Serum Albumin C Small molecure 1 Test Uppsala Now 2005 Modified Modified By 1 immob of enzyme amine pH4 5 2106 Run 2006 03 27 15 59 Import user 3 assay 2105 Run X 2006 03 28 2 09 Import user Name Preconcentrat ion Description OK
53. buffer 1 Select assay step 2 Select where to define variable values for each amp ssay Step Method C Define all variable values at run time Overview Define all variable values in method Define some variable values in method and others at run time General Settings 3 Enter values for amp ssay Step pH scouting Assay Steps Sort in alphabetical order 1 _ Fowcell Flowcell2 Fowcel3 Flowcel4 lame Name Name Name Cycle Types 0 60 pg ml CA pH 4 0 30 4 0 Sample amp Assay Setup I Ml 0 ug ml HSA pH 5 5 15 uig ml HSA pH 5 5 60 ugiml CA pH5 5 30 ugiml 5 5 F K 2 i A A X i m M i Verification Verification amp Setup Run Sample amp Assay Setup No assay step needs data Sample amp Assay Setup Cycle Run List Cycle Run List Biacore A100 HC 2 X 2 18 No i 1 pH scouting 2 pH scouting
54. Biacore A100 Control Software Biacore A100 Edit View Help Method Builder Run O pranm demo i MAb screening with RAMFc capture 050812 2045 Serum Albumin Description Created 2005 08 12 11 57 24 Created By Import user Started 2005 08 12 11 57 27 Started Import user Ready 2005 08 12 17 35 11 Instrument 20015 Number of Cycles 17 open i Biacore A100 il E X Surface Preparation pH Scouting Assay Development be 71 Screening 71 Kinetics and affinity _ Concentration Biacore Methods gt Surface Preparation pH Scouting Assay Development Assay Conditions Regeneration Scouting Screening Screening Screening with capture Kinetics and affinity Kinetics and affinity Kinetics and affinity with
55. CAC2Dug ml pH5 Fclil 3 corr E nam run sims E m Hum cM c c ri M NE MM 7 Auai M MEE m ME D e mi NE we crc o WE DER HE aM reped c NEN NE MN Parameters Biacore A100 alidahinn info Sensargram Table Parameters l Conc D Initial Values 0 0 Validation info Sensorgram Table Parameters Conc 0 Initial Values Residuals Initial Values Validation infa Sensorgram Table Parameters Conc Initial Values Residuals Initial values For fitting ka 1 00 5 kd 3 178 kt 5 EEer7 Residuals Kinetics Biacore A100 208 9 T 2 f 55 RIET 5 Validation infa Sensorgram Table Parameters Canc 0 Initial Values Fesiduals Residuals 2 1 1 9 2 a 3 4 5 T Time s Next gt
56. Setup Run Method Setup Run ad uo allie 25 C sample time sz enteraj ILnmhmnl sample t Tonry Samp Sample 1 times ac entered Before every 20 cycles m Ta Samphe T 120 sr 120 s pinire lor E Pemmwalmm 1 104 HaDH 32 L 1 1 Biacore A100 94 5 Sample amp Assay Setup Method Sample amp Assay Setup time Define some variable values in method and others at run time 2 Define all variable values at run Setup Run Cycle Run List T Sen RU b 2 Enter values for Assay Step Sample Sorin alphabetical order Sample 1 meweman mswems Name Conc Mw 0a Name cone um MWiDa Name Conc um Mw Da Name cone um mw E Name 27 Conc uM MW Da 09 10
57. Biacore A100 3 31 Setup Run Tray 1 S6 Welburopae eero esgent Insert Trays Start Run Modified Modified By 1 immob of enzyme amine pH4 5 2106 Run 2006 03 27 15 59 Import user 3 assay 2105 Run X 2006 03 28 2 09 Import user C Antibody J Concentration demo CJ DNA DNA C liposome 1 manual C Serum Albumin C Small molecure Test C Uppsala Nov 2005 Name Preconcentrat ion Description Cancel OK Biacore A100 32 3 3 2 2 pH Sensorgrams by Flow Cell 519 Fc2 pH scouting 100 120 Fed pH scouting Biaturilnrirumenbof ne Anari temperatus Teas tarnparatuna tatinamard stale 3 d 2 Curve start at zero
58. v gt 2 biacore LED 272 ready temperature 2 1 2 ZJZ Y DEEANN h L EKOE Y h INFJULRULAICTLILDORGDE AEK v 5 PPF 285000780 Water 2 2119880 I Biacore A100 2 7 1 Tas Water PIA A nn so 2 3
59. Evaluation Items Binding levels Jindinq evel Evaluate Data einn Save And Finish Biacore A100 54 gt 2 55 4 2 100 4 55 4 2 Regeneration 30 1 CECD
60. ey Type Approved Data j B 2 PITIE RDIR ype Evaluation Navigation panel 25 Select Data Evaluate Data Screening EN ORI NU Kinetics u N Affinity c I EIE Concentration Biacore A100 9 1 BIAcore A100 Evaluation Softoware Start gt Biacore A100 Evaluation Biacore 100 Evaluation Software User name admin Fassword BIACORE A1OO mama User name c Password Eek File Edit View Help Assay items Information Select Data _ Name Type Status Modified Modified By Fold Concentration demo DNA DNA Serum Albumin 8 738 hlished Small molecure m upisne j Test Uppsala 2005 Administration Actions New Summary Folders
61. 9 2 2 1 Kinetics and Affinity Kinetics and Affinity with capture lt gt Biacore A100 202 9 T 2 Evaluation Items Evaluation H Approved Data Items EHE Evaluation Items Kinetics1 All curves Biacore A100 9 T 2 203 Add All As Single gt Add gt Selected curves and combinations Next gt i xEH Biacore A100 Evaluate Data X Examination Eralualion liens 00000 00 5 22 Azosulfamide So Dieci Fzl 1 3 000 Carbone du Sata Fol5 3 ou
62. Import Em BARFE 50809 2051 Bun Completed BE 5 17 PM Version 3 Ef Subclass Determination C Hub 22 Run Compeed 12 581 kinetics Furosendde 5005 2023 Run Completed 5 57 m MAS screening Tr EE L2 2084 Run BI127200S 5125 PT E gH arini HSA anal CA ES Bu 471005 30 03 T rening Serum Alursns 50807 20056 Bun Compl ed 7 05 Bm Sereen Small Hoces 5060 2087 Run Completed 632005 1 51 z aite uz CE HEAR Run Cumrlered TIS PT Navigation Left panel Central panel Right panel panel utku Btatuz Insiramunt ofina Analysis temperatur Tray temperature mesas Status bar Kaj ES Status Inetrumaent 20102 online Analysis temperature 25 0 25 0 C Tray temperature 15 0 1 5 0 C Inztrumant state No sentor chip inserted idle REC PC Biacore A100 2 2 2 2 2 1 gt Toobar 007 7 2179 5 l ACER Temperature Analysis temperature e 4 40 COD8BBH L
63. t x et p L tammi pnas a u m Analysis tamosrahrni 25 0 2510 Biacore A100 16 2 d OK Bad Bad gt Change buffer 1 4 146 l Hydrodynamic Addressing 150 Bad Spot Configuration 153 Hydrodynamic Addressing Biacore A100 2 17 3 1 V QM e 6 pecu Series S Sensor Chip CM5 53174 BR 1006 68 Series S Sensor Chip CM5 certified 5314 BR 1005 30 Series S Sensor Chip CM4 certified 5314 BR 1005 34 Series S Sensor Chip CM3 certified 3 BR 1005 36 Series S Sensor Chip C1 certified 3 BR 1005 35 2 DNA Series S Sensor Chip SA certified 3 BR 1005 31 3 1 27
64. 192 9 Data selection Fe Edi Hip ResultPlot1 Evaluate Data Settings 4k Plot 4b Resunt G Report point weas variable E was Repot point dilfenenk curves E valualion Tiso repost por utsut variable Daa lere E Z Eaumlinri liere Ranking binding be i Ranking binding sh 1 Lemrertesd bed Cerrecied Assay Step Purpose Make following columns available in result table Make following curves available Next gt Biacore A100 9 193 Next gt ResultPlot1 Settings 4 Plot 4 Result Biacore A100 18
65. Biacore A100 HE21 FITS Settings Enable automatic data validation Evaluate Data Settings Preparation Examination Results pre Daa ama E E Fens i Enable automatic data validation Selechad Cures and curva combinations Add Single gt Remave All zRemeve Biacore A100 220 9 Validation info gt kinetics Der P 2n e Y Fc1 4 3 corr HSA Furosemide 25 Fc1 5 3 corr HSA Furosemide 25 Series 3 Series 4 MN I W F gt L 1 4 3 HSA Warfarin 25 Series 7 Fc1 5 3 carr HSA Warfarin 25 Series 8 Affinity Fc1 1 3 corr Furosemide 25 Series Fc1 2 3 corr CA Furosemide 25 Series 1 2 Fc1 4 3 corr HSA Furosemide 25 Fc1 5 3 corr HSA Furosemide 25 Series 3 Series 4 1 4
66. de 9 bo U PBS 6 DMSO E 1 Running buffer 84 PBS 5 0 50 4 5 ti Biacore A100 6 127 DMSO fien DMSO 5
67. Please place Tray 1 onthe camage and press The hotel hatch wal be clased aubonmalicall cer miube Tn action t 1 Reagert plate 7 II D E 8 Wel T 0 C G I OC YC X X YC 2 JOOOllO e e e x x e e e sC OC OC C COCCO Oe e e e ee ee ee Jee eee eee 00090909 Remaining tme before the hotel hatch wil close 58 60 OK 19 5 Run gt Biacore A100 2 15 Liu J E ELS Docking Hydrgodqnangc addressing with normalize rhided Biacore A100 Hydrodynamic addressing Result run Hydrodynamic Addressing 2006 01 18 14 21 21 User 10 adrminiglralar Run Daia 14 20 Software A100 Fnnlrnl Sofware Varsinn 100 ID 20102 CenfiqurationlD IFCTD4 Amalyze Temp 25 D C Respenze i znna ELT 2 ELZB 2177 Zl amp 7 2150 210 ELE 1 E
68. Biacore A100 ReJ Affinityl renoarati n Examination Results 1 Fe a iot Faisenesel Senes Table KD M Rmax1 Biacore A100 212 9 6 Furosemide1 Response 2 0 2 6 4 6 1e 5 1 2e 5 Conc M KD 8 231 7 Rmax1 11 92 RU Iffse 0 7334 RU ChF 0 04304 2 Validation info Sensorgram Table Sensorgrams Parameters Conc 0 Initial Values Entry Validation message 2 Note Data quality seems acceptable 1 Note Crossvalidation results KD 8 231e 7 S D 2 881e 8 Validation info Sensorgram Table Recalculate Modified Results
69. Delete Move Up Move Down New Immobilization Wizard Chemistry Flow cn 1 DpH Contact me mim Z Lise spot 1 3 10 FZ Usern liid Amire Biacore A100 Methodology Handbook Surface preparation Biacore A100 46 4 4 crude
70. posliri Sample 1 stet Curve e Ligand C Adusi ta C Fe ampie C Cede Fe Ligand Sampe et repost point position S a Dus selection Mak Cuckrs gra laba caves Eu Fr1li5 I 5 Li 1 Condborig MK Hace r 1 1 ah Autres 2 Ma Buffer E i _ r 3 Na DEBE pure sm 4 1 Rabbit serum mh hte LI Suwtup Bufe _ NaN 3EBE Buffer r F 5 Suet up GERE Buffee 4 z Rak z T Famitrnl sa contra 1 rontrr 2 1 z 1 Raebbt serum sb Antin n Control sa Meg centro 1 contr r Fidis 2 9 ab Active F 9 50 Warfarin FR j 2 jRaseamas Action sa B Fi r sinud r Fc3 5 5 samm alb Action ii Sample Digitasin 5 Digibusin r Feliz 3 2 Gurea serum sib 12 Sample Warfarin 50 Warfarin Fr3i1 3 1 Donker swum sb Actie le 13 sample Msproken 2 Fe1i1 nrr 1 1 Donkey serum alb Corrected F 14 Sample Phrenytoin Phenytoin e Fed aamir 4 2 Ratseranm alb
71. 42 3 8 New Immobilization Wizard hi pepa non Dora petim alicn al bersperaluses baes fan A E T Make sane that and Adeang hae ran pi 2 dd and megen bo the rpecifiad po ma Seal wath lcd Fabra Far eare 4 Pise pisie an tha Isa and miesi inta the ins lamen 5 Fil bords risate rn buffer and boites 1 4 valh buffi fo ad er Puls knee wei 1 cd buller be palomed Hake sure that the mash the baton ol Pres boies Emolp Fe v ila cord ane TZ re roten for cantu Sal nun mus 4 Cem Custom Chemistries Biacore A100 3 43 Be ce Create Amine E L lt ld Surface Thiol 5uFTac Thiol l Biacore A100 44 3 Custom Chemistries Injection sequence
72. Fitting status Done A100 9 205 Accept Current Abort Current Ost S Abort Remaining i Kinetics 1 Evaluate Data Settings 4b Preparabon Examination Results Fralaglicm rms i Azosulfamide kd B Zia 174 Hina 45 77 RU Ded 1 12 ED 2732 L31937 Foiz 3 con ator ankesa padari 10 Table Tinne Rmax1 2 1 Accepted OK Carbonic anhydrase Azosulfamide Fcl 1 3 corr UTC TREES 1 3 Accepted OK Carbonic anhydrase Azosulfamide Fc1l 4 3 corr 0 1777 Du 7 956e 3 2 713 4 2 833 7 4 Accepted OK Carbonic anhydrase Azosulfamide Fc1 5 3 corr 0 3274 1 758e 3 2 565 4 3 024e 7 __5 Accepted OK Carbonic anhydrase Azosulfamide Fc2 1 3 corr 0 2081 09 8004e 3 2 952e 4 2711e7 6 Accepted DK Carbonic anhydrase
73. 3 35 8 Cycle 1 2 Cycle 3 4 Cycle 5 Cycle 6 Cycle 7 8 NHs 1 2 4 5 NHS 2 4 1 5 50 60 pH NHS 10 mM pH 4 5 150 mM NaCI 0 0596 surfactant P20 2 4 75 85 1 Biacore A100 36 3 3 3 1 New Method v Surface Preparation Immobilization New Immobilization method OK Setup
74. 2 3 4 Spot Usage Ligand Contact time fmin 1 Actidaeact Immobilized LT E w 2 4 ct deact Immobilized BEE Act deact 3 Immobilized Act deact ZU v FYFE Unmodified Unmodified Biacore A100 38 3 3 3 1 1 1 1 1 Flow cell 1 Chemistry Spot Usage Ligand Contact time imin Usespoti 3 Amine coupling 10 min activati v 1 Immobilize for captur GST proteini 156 Use spot 4 5 LT CO 2 Act deact BEN 22 p A JJ Overview 2 7 W x Chemistry Amine coupling 10 min activation Flow cells 2 5 pots 1 3 ZUR S Is x Ligand injection Contact time min Spot Flow cell 1 Flow cell 2 Flow cell 3 Flow c i 1 15 1 GST protein butter butter butter 21 J 214
75. 2 2 Command Sample1 Commands 8 Capture Sample M T Insert Command Remove From Cycle Flow rate 30 prin Move Contact time lgn s Move Down D issociatian time 60 s Regeneration 1 i 1 Type Sample Normal Flow rate 30 Contact time 120 Dissociation time 120 Biacore A100 108 6 Regeneration 1 Commands Capture Regeneration nsemicemmand Regeneration solutions Regeneration F Remove From Cycle Regeneration Fe2 Flow cell 2 RegenerationFc3 Flow cell 3 o Mewelp Regeneration Fe Flow cell 4 Move Down Contact time 30 s Atleast one high viscosity solution Regeneration solutions Flow cell1 Flow cell2 Flow cell3 Flow cell4 B3 ES lt A 7 10 mM NaOH Contact time 30 Biacore A100 6 109 D Z Command variable 1 Method Variables Evaluation Variables Method Variables Evaluation Variables Fc 1 Mame De Conc Numeric Fc2 Name Fc3 Mame Molecular weight Numeric Fc4 Mame Time s Diss Time 8 Flow ul min User defined variables Add lelete Evaluation pur
76. Sample Carry over Insert Command 97 212 229 5 Commands OOOO LH Capture Sample Insert Command High Performance Kinetics Remove From Cycle Flow rate 30 ulmin blove Up Contact time 0 Move Down D issaciatian time 0 s ample 1 amun 1 High Performance Kinetics Flow rate 30 Contact time 60 Dissociation time 60 Biacore A100 120 6 cycle type Variable Method variables Evaluation V arrables Predefined variables Conc Numeric Molecular weight Numeric Evaluation Variables Conc BE Assay Steps J timer ss A ample eerie L Sangala tiri as erdarad A Sample tias 55 entered Base settings Name Sample seres 2 Purpose sample Lonnectto cycle type Sample n umumummumumumummu M nt Connected Sample Solvent carrection on Number of replicat z Londitioning C Randon i a time Assay Steps
77. 34 3 3 3 1 5 HA 5 1 152230 2 52524 3 Flow cell 1 Flow cell 2 Flow cell 3 Flow cell 4 000100 00610 AL 080100 Cycle 1 x Cycle 2 11 itt 11 1 2 iz DN Ep PENA Com Ethanolamine spot 1 2 Ligand 2 spot 1 2 w RU 60000 Ethanolamine spot 1 000 55000 Ligand 1 spot 1 10 U 00 s e n a 50000 45000 40000 A T A 1 EDC NHS spot 1 2 DU 00 2 SO9UUU LLLLLL Response 30000 25000 20000 r r 500 0 500 1000 1500 2000 2500 3000 3500 4000 5 Biacore A100
78. 45 55 5500 45 PIRIAI2 50 5 mM 50 Na SD NanH CEST mm _ mw Blatut Insinament Anahi Trapternperature m gt pH pH NHS Adjusted sensorgram a pHA O soon pH4 5 4000 pH5 0 5 4 0 2 2000 4 5 DIT pH 5 0 CATLETT 1000 pH 4 0 CC A SUA pH pH pH 5 0345 pH 40 SA R espor ea 1000 2000 50 50 100 150 200 250 300 Time 0 Sample 1 Start s Biacore A100
79. 0 S Biacore A100 3 33 Run Information Fun Pumamg lemo scoutng HSA and CA 2085 Run zladus Corsplelad Sitat Hime PDT T 3c 33 55 Stmrbed ba urere Stop time TIT Ciaked ber use Duration 112213 Flow Inztramenk 20015 Humheralcgclez 4 e run list s EE uw 5 ple Fc rj ample Fn 5 F 1 HSA pH 4 158ug ni HSA pH pH 4 0 30 goal pH 40 2 MEE EIE pla 2 pH 4 5 Wapi DA pH 45 EN ri 30 ppm HEA pH 5 pH 50 Q pH EO noil DA pH 5 0 4 pH eating HSA pH 5 5 ISun HZ amp pH 55 2 pH 5 5 D pH 5 5 EHEEHEEEHEEEHHEEHHEEHHEEHEHEHENHEHEHNHEEHEHEEHEHEHEEHEHEEHEHEEHEEHHEEHHEEHNHEEHNENENEN Assay steps data Viials and plates HEB EP Position Content Fel vii ContemtFcd ContentFcd volume 90marmlHSApHAO 8ug niHSA pH 40 BO po ni CA pM 48 46 DH 45 EO pl pH 4 5 P 45 0
80. 185 16 l Save and Continue Folders Small malecure 3 items E Antibody CA Azo kinetics Run Completed 22 17 mm user 3 Coe EE NR d 1007 03 13 11 02 acmiristrato dw x E dema A zi 11 02 an DNA DNA 7 immobilization instruction 7 liposome manual serum Albumin Small malecure mM G me Type Approved Data Status Finished Biacore A100 170 9 11 Report Points UView Sensorgrams RU Adjusted sensorgram Pos control2 10 uM 35 2 Q9 30 50030 25 5 m4 20 a 15 2 9 g0 amp F ss se S 5 D SER B APER F Time
81. Positioning Trays Start Run Biacore A100 4 HS 63 4 2 2 Biacore A100 Evaluation T A Ege eb Open Assay Development Regeneration Scouting Regeneration scouting OK eg s Notebook Close L Next gt L IN9 S Tables Quality Control Save And Continue PORA SE Notebook 3 Close Biacore A100 64 HR TERHRIGE OD 7S 600D2E T HIS Evaluation Items Response levels Evaluate Data Fslaliom tems humhrais a amp 7 Fell
82. 7 3 7 3 1 Normalize Dock 10 BlAnormalizing solution 70 glycerol 28 2 k 77g RECEA S Normalize Normalize Setup Run Maintenance Required solution Bl amp normalizing solution glycerol Click Hest ta continue Biacore A100 admin Biacere 4100 Canliral Hun Fia Wem Tons Bun riska ch uj m Chp Temp Rack Tu Setup Run Trap 81 Boi o 380ul BIAnormalizing solution Bottles 8 43 run duration 11min x 100 el excludi botte dead volume and standby consumption 330 miiday 100 ml excludryg bottle desd volume and standby consumption 95 miid IO ml excludryg botte desd valume and standby consumption 10 ID excludrg battle dead valume and consumption 72 LOO excludrg battle dead volume and standby consum
83. BIAdesorb solution 1 0 5 SDS BIAdesorb solution 2 50 mM Gly NaOH pH 9 5 BIAdesorb solution 1 4 4 Desorb Desorb Setup Run Pinna Required solutions from Maintenance Fit Bl amp desorb solution 1 Bl amp desarb solution 2 NOTE Do not run this procedure with analisis rack temperature below 20 Click ta continue Next gt NUES Biacore A100 T 137 Desorb Positioning Setup Run Tray 81 Reagent piste J m wel Ricroplale OOODGOCOGOOGOGOOCQOCQ QO ee 1900000000000 00000 000006 900000000006 0 00000 00000 BIAdesorb solution 1 BIAdesorb solution 2 3800ul 1500ul Next gt Biacore A100 Insert Trays Run gt Desorb Standby flow
84. Series S Sensor Chip HPA certified 3 BR 1005 33 Series S Sensor Chip L1 certified 3 BR 1005 38 4 His tag Series S Sensor Chip NTA certified 3 BR 1005 32 Biacore A100 18 2 4 96 384 1 R1 2 AH 1 3 EH 1 1 0000000000 1 100 2 19 25 1 Locked
85. 10 Biacore A100 Instrument Handbook Biacore A100 Software Handbook Control software Evaluation software Biacore A100 Methodology Handbook Sensor Surface Handbook Biacore A100 151 89 ms sess CES rine _ TOSS vm i 525 576 D DR gt On gt lt
86. Tris Glycine 1 Id vts Ue Biacore A100 24 3 5 200 ug ml 10 mM CHELE SD pH 26 pH 100 ug ml 10 mM Disodium tetraborate pH 8 5 1 NaCI Biacore Biqcore A100 Methodology Handbook cca Biacore A100 3 1
87. a Do you want to change buffer 1 4 before starting the run HC 2 LLL 75105615 No i Setu p Run Reg condition 1 buffer buffer buffer mouseIgair2ugml Reg condition 1 buffer buffer buffer mouselIga zugrml Reg condition 1 buffer buffer buffer mouselIga zugrml kh Reg condition 1 buffer buffer buffer mouselIga zugrml Reg condition 1 buffer buffer buffer mouselIga zugrml Reg condition 2 buffer buffer buffer mouselIga zugrml Reg condition 2 buffer buffer buffer mouselIga zugrml Reg condition 2 buffer buffer buffer mouselIga zugrml Reg condition 2 buffer buffer buffer mouseIga zuugrml Reg condition 2 buffer buffer buffer mouselIga zugrl Reg condition 3 buffer buffer buffer mouseIga zugrml sample amp Assay Setup Reg condition 3 buffer buffer buffer mouseIga zuugrml Reg condition 3 buffer buffer buffer mouseIga zugrml Reg condition 3 buffer buffer buffer mouseIga zuuarml Reg condition 3 buffer buffer buffer mouseIga zuuarml Cycle Fun List Reg condition 4 buffer buffer buffer mouseIga zuuarml Reg condition 4 buffer buffer buffer mouseIga zuuarml Reg condition 4 buffer buffer buffer mouseIga zuuarl Positioning Reg condition 4 buffer buffer buffer mouselIga zuuarml Reg condition 4 buffer buffer buffer mouseIga zuuarml Reg condition 4 buffer buffer buffer mouselIga zuugrml
88. 02 25C 8 Spot Configuration Spot Confiquration Setup Run Use 5 buffer as running buffer Click ta continue Next gt Spot Configuration Setup Run Ver TERT SIE The Restore last settings restores the last valid Spot configuration only to be used when the Spot configuration has been aborted Spot Configuration Restore last settings Spot Configuration Next gt Biacore A100 154 T A100 Spot Configuration Result run 101 Spot Corfiguratice 2006 03 24 20 34 27 User 10 adminislralar Rin Date 24 20 24 Seftware 10 AT Control Sofware Sarsion 1 Instrument ID 20102 Cenfiquration ID IFCTOA Analyze Temp 250 Previous center row New cemer row 157 157 214 214 FO 31 anu QUALITY CHECK stance number or detector rores hatwasmn call centars FC2 FCl 57 B 6 2 OK sa 57 2 E3 uv Selected center row Max relative response 2 2800 amp lt 1400 18485 160 amp FC l 02 2027 2037 2035 1834 17E5 Linear regr
89. i buffer 2 EE 4 TT VON ONT ET p cua S s E BL E 5 buffer 42 0 d d alc MEE e 8 es a 5 x 2 e UN UT OD MEC E Sample series1 Biacore A100 6 123 Verification Setup gt amp Assay Setup Cycle Run List Cycle Run List Startup Buffer Buffer Buffer Buffer Startup Buffer Buffer Buffer Buffer EE M Sample series 1 protein X buffer buffer buffer 0 5 Sample series 1 protein X buffer buffer buffer 0 5 Sample series 1 protein x buffer buffer buffer 0 002 Sample series 1 protein x buffer buffer buffer 0 004 Sample series 1 protein X buffer buffer buffer 0 008 Sample series 1 protein X buffer buffer buffer 0 016 10 Sample series 1 protein X buffer buffer buffer 0 032 Sample series 1 protein x buffer buffer buffer 0 065 a2 Sample series 1 protein x buffer buffer buffer 0 13 13 Sample series 1 protein X buffer buffer buffer 0 26 kr T ed ps TETTE EE E mumununununun 15 Sample series Z Compound X buffer buffer buffer 0 16 Sample series 2 Compound x buffer buffer buffer 0 15 Sample series z Compound X buffer buffer buffer 0 31 18 5 series 2 Compound x buffer buffer buffer 0 62 14 Sample series z
90. 4 OL 46 na aUa teet SSS sss 47 OO 47 TEENS ha hanaq 52 A 55 56 e ju 4 8 RN 57 4 2 2 Re 63 5 66 66 425 2 meet P 71 73 5 Pi EOD US RR 74 ODE d NR Emm 75 5 2 100 AME ZUSID a 102 E 102 6 2 FEBRE E E a WA R 115 6 3 lt 124 6 3 1 DMSO esI 128 5 Gc A 132 gt 134 NR 136 DEO RD 136 7 1 2 Deso G Ord 139 yep encor Ns i 143 145 eos debe toam 146 esk Chande bune RU 146 SCC QA SN 146 5259 A 146 r s SOrD Cr 147 Biacore A100 DONE 48 005
91. EZ Tool bar J 27525 Chip Information Chip name 2005 06 07 11 00 59 First dock date 2006 08 07 11 08 Chip type Last immobilization 2008 08 14 11 34 Lot number 1160109 Ligand RU State Chemistry Run Name Time Immobilized By miga 10771 Amine coupling 10 min activation surface stability immab test 1 2006 08 07 19 40 administrator mig 10684 Immobilized coupling 10 min activation surface skability_immob best 1 2006 08 07 19 40 administrator Unmodified Amine coupling 10 min activation surface stability immob test 1 2006 08 07 19 30 administrator Hot used used Immobilized Amine coupling 10 min activation surface stability_immob test 2 2006 08 11 17 14 administrator Imrobilized Amine coupling 10 min activation surface stability imrmob test 2 2006 08 11 17 14 administrator LinmwodiFied Amins coupling 10 min activation surface stability immob test 2 2006 08 11 17 14 administrator Not used used Immobilized Amine coupling 10 min activation surface stability immob test3 2006 08 14 11 34 administrator Immobilized Amine coupling 10 min activation surface stability immob test 3 2006 08 14 11 34 administrator Unmodified Amine coupling 10 min activation surface stability immob best 3 2006 05 14 11 34 administrator Hot used Not used Hot used Hot used Mot used Hot used Mat used
92. 96 Reagent plate 96 Well Microplate 384 4 2D Z7 UL 8S Z v F Dese TVCU SIS Reagent plate 384 Well Microplate Empty Biacore A100 2 21 2 2 3 x Tool bar Che The sensor chip will be ejected OK j 10 Biacore A100 22 2 2 3 Assay Folders Search New Folder ok Cancel Name Ok Assay Biacore A100 3 23 3
93. Rabbit secum ah Ru serum ado Puman senm sib adie seun ab simui seram al Artes Farini seua ad Arten ule pig serum Donkey seum sh Pankey serum alli Corrected Rat seran alli Correcked Rabbit serur alb Corrected Donkey serum alb Corrected Porcine senum all Corrected simum all Corrected Rahhil seruis alls minga piq eerun alls Corrected Duineca pig serum als Corrected Human seram alls Corrected Porcine senu all Rat serar alli Corrected contre Meg F r si nd Digitasin arfar Haprowen Pisenyboim anilana conr contre Warlarin ut urn centro rari arim Fur send Digitasin arfann Beg conten Meg contro warfarin Furosemid CESA Furnsemsl puer Cemtenl sa Meg contra m I Feli 3xurr lt FT 1 BrnrF oua CU Ln Fcl i5 doxurr Fr4 5 4 xurr Fc axar Fci 2 3xon Fc32 3xnmm Feis 4ocurr Feds 4xur Fea EE CEP 187151515151 lt q lt lt xp pp xr xp xL l lt lt 1 T 17177 Tt X MA QA MI FG om xA RE Mm m ra up gain oe gi cp 1
94. Regeneration 1 Sample amp Assay Setup Verification Setup Run Sample amp Assay Setup i Biacore A100 4 69 sten Setup Run test Sut n alhsbeical order x Senpe amp Assey d P 3 P zleri F fictio 17 7 AJJU Cycle Run List Do vau want ta change buffer 1 4 before starting the run HC 2 V LLUGBInmOS38 532 730 ES No Biacore A100 70 4 Cycle Assay step name Sample Sample Fc2 Sample Fc3 Sample Fc4 Conc Conc Fc2 ConcFc3 Conc Fc4 Setu p Run He Ligand activity test buffer buffer buffer ProterinA 2 Ligand activity test buffer buffer buffer Proterin 2 Ligand activity test buffer buffer buffer ProterinA 2 Method Dem Ligand activity test buffer buffer buffer Proterin 2 Ligand activity test buffer buffer buffer Proterin 2 58 Ligand activity test buffer buffer buffer ProterinA 2 Ligand activity
95. Save Method Save Method As Setup Run Overview Sample amp Assay Setup Biacore A100 6 113 Sample amp Assay Setup setup Hun Cycle Run List Biacore A100 114 6 Cycle assay step name Sample Fci Sample Fc2 Sample Fc3 Sample amp g4 Conc Fci Conc Fc2 Conc rc3 Conc rcs Mw Fet mw Fez mw FG Buffer Buffer Buffer Buffer Buffer Buffer Buffer Buffer St a rtu p 2 Control Sample 2 Buffer Buffer Buffer Buffer Negative Negative Negative Negative 5 Control sample Positive Positive Positive Positive A1 A13 A25 A37 10 10 10 10 2 A14 A26 A38 10 10 10 10 15 27 A39 10 10 10 10 44 A16 A28 A40 10 10 10 10 A5 A17 A29 A41 10 10 10 10 6 A18 430 442 10 10 10 10 A19 A31 A43 10 10 10 10 8 20 A32 A44 10 10 10 10 49 A21 A33 A45 10 10 10 10 A10 A22 A34 A46 10 10 10 10 SQ m ple 20 D 811 23 A35 A47 10 10 10 10 A12 A24 A36 448 10 10 10 10 449 A61 A73 485 10 10 10 10 450 A62 A74 486 10 10 10 10 A51 A63 A75 A87 10 10 10 10 A52 A64 A76 488
96. fi B name Next gt i Biacore A100 188 9 cyclo a as number Digand Curve marker cone Sample 7 1 154 2 7 Donkey Neg control 3 corr a serum alb Fc1 2 u 459 1 8 7 Guinea pig control corr serum alb ly Fci 5 389 8 7 Porcine control 4 serum alb Z 2 1 835 4 7 Rabbit Neg control 3 corr serum alb 7 FC2 2 317 2 7 Rat serum Neg control corr P alb 7 Fc2 5 a 111 2 Human Neg control 4 serum alb r4 7 FCc3 1 i 45 9 Donkey contral 3 serum alh ly T 2 u 572 9 Guinea pig Neg control 3 corr s serum alb Fc3 5 a 295 0 7 Porcine Neg control 4 serum alb 7 Fc4 1 684 9 7 Rabbit control 3 corr serum alb gt u 194 1 Rat serum Neg control corr alb 7 Fc4 5 1 2f Human Neg control 4 serum alb 8 Fr1 1 251 3 8 Donkey Neg control corr serum alb TEN s N E 52
97. gt lt gt i D i Biacore A100 5 85 Assay Steps e gt v 7 1 Create a new cycle type select an existing From the list Cycle types currently method Sample Sol carr Delete Conditioning Copy Rename B New Delete B Insert Command Commands Commands Capture Insert Command Regeneration Remove From Cycle Laru wl Solvent L arrectian Move Up I3eneral Move Down Move Up Move Down 7
98. in pHs Fol 2 HSAN Mogin in pH5 1 m m m m m l 4 k Lowest riur CB 189 17 Ranking binding levels Y binding late Ranking binding stability X stability early Binding vs stability X stability early Y binding late Biacore A100 Controls sensorgram 0 Samples sensorgram 194 18 Tools Color By E Label Caption Label Ha label onc sample Caption mmm G Color d Nea Label By e name
99. Biacore A100 T 139 7 1 2 Desorb and Sanitize 1 1 JD 82 3 BlAdesorb solution 1 0 5 96 SDS BlAdesorb solution 2 50 mM Gly NaOH pH 9 5 e BlAdisinfectant solution 7 5 ml 100 ml Desorb Sanitize Desorb and Sanitize Setup Run This pracedure iz divided inta Four steps two Liesnrh steps steps 1 and 2 Sanitize step step 3 Wash step step 4 Hun time about 1 hour followed by a recommended standby time of 3 4 hours Minimum required solutions from Maintenance Bl amp desorb Solution 1 30 ml BIadesnrh Solution 2 SU ml BlAdisinfectant solution diluted according to Instructions For Usel 105 ml Do nat run this procedure with analysis temperature below 20 Click to continue Next gt 2 789077 1 Desorb and Sanitize Setup Run Insert all tubinigs Im minimum 90 ml Bl amp desorb Solution 1 Click Run to start the procedure NOTE Do not abor
100. General M E 1 Method Variables Sample 1 Fc2 Name Fc3 Name Fc4 Name PAN T Contact Time s Diss Time s Flow ul min 0 2 Evaluation Variables BETA DO IZICBBReTUSaqosEO ORUM EBJOSE Conc Evaluation purpose General Biacore A100 90 5 Show advanced settings BIZY FCIZAMFZ AASMESGE T2XSL2Uv2dg3 3 Advanced Extra wash after Flow cell 1 Flow eell 2 Flow eell 8 eell 4 r Stabilization period og before injection Stabilization period after injection Extra wash after injection with 50 DMSO Stabilization period before injection 1727 21030800455
101. 1 Capture 2 Sample 3 Enhancement 2 Biacore A100 86 5 4 Regeneration 5 Carry over 30 mim 303 B 4 7 24 5561881 6 Solvent Correction 30 ul min 30 7 General Sample Sample Regeneration Capture sample 6 Biacore A100 5 87 C BIY KORE
102. D Sample Insert Command sample1 Type Normal Flow rate 30 Contact time 30 Dissociation time 30 Biacore A100 130 6 Assay Steps Method Assay step 1 Semple Not connected 1 time as entered Connect to cycle type DMSO check sample amp Assay Setup Method Assay step 1 25 7L D JL 1 2 Name High 3 4 Low i Verification Setup gt amp Assay gt Run List l Biacore A100 6 131 Biacore 4100 X 2 you want to change buffer 1 4 before starting the run gt saa d 27250 Yes Positioning TraysStart Run gt Flow cell1 2 Flow cell3 4
103. 5 LAO Sample amp Assay Setup 22 73 705595 Method Method 0 lt lt D 2 Setup Run Save Run Method Setup Run Methnd Setup Run SetuD Run
104. Biacore A100 Desorb and Sanitize Meienance The gsnrh and 5anitize procedure is completed Allow the system to run in standby made far at least 3 4 hours before performing a run 5 Standby flow Biacore A100 T 143 7 1 3 Superclean Desorb Desorb and Sanitize 1 30 e 1 acetic acid 0 2M sodium bicarbonate 6M guanidine hydrochloride UE 3 5466 amp a 5096 DMSO 10 mM hydrochloric acid 1096 DMSO 0 22 um k gt 40 50C Superclean ouperclean Setup Run This procedure cleans the flavi system Required solutions far protein work 1 acetic aci 0 2 M sodium bicarbonate guanidine hydrochloride 10 ml hydrochloric acid Required salutions For small molecule work 1 acetic aci 0 2 M sodium bicarbonate 0 dimethyl sulfoxide 104 dimethyl sulfoxide Place
105. EC e General Setting l 25 RESAS E L RET 920 027120 96 Assay Steps Remove Step ismga S mes entered Sample 1 1 Sample 3 times as entered Sample L tira entered A Sampe series 2 ample Sampit L tini as enbened Biacore A100 6 117 Cycle Types lt protein X DRIER EREI 5 AB 1 Create a new cycle type or select an existing from the list Cycle type description Cycle types currently in method Mew This cycle is used in startup sample and Includes injection of sample regeneratia ARMIS T ROSA Solvent correction Delete Conditioning Rename 2 Insert the required commands 3 Configure settings Far each command Commands Capture Sample Inzert Command High PFgrfarmarce Kinetics Remove From Cycle Flow rate 30 Move Contact time 1 20 s Hoc Donn Dissociation time 20 s 1 Regeneration 1 arry over 1 Type High Performance Kinetics Flow rate 30 Contact time
106. T X There is at least series that should be inspected vau really want to proceed Mo Yes i Finishing and lacking kinetics evaluation Kinetics Save Save As 214 19 216 20 gt Biacore A100 9 209 Evaluate Data Evaluate Data Evaluation Items Affinity1 All curves Curves and combinations in affinity evaluation Next gt Biacore A100 Affinity Furosemide2 Fe41 3 Laibonsc any Fur seridez Senes Next gt
107. m EILAN QU 5 35 5 3U 5 JALAN 177510 80 CASE d RIN 9 Ab K IC 60 2 B S 7 B 7 2 Fisch CRUS VN i x iB
108. serum amp lbumin e Small malecure 1 Test Evaluation Stqtus Finished Biacore A100 2 RU Azosulfamide Azosulfamide Export Curves 1 Scale 25 1 Giridlines a Legend Graph x k 15 Gridlines Legend 2 NN dM 100 100 200 300 400 500 TD HH Time amp Azosulfamide 35 Scale 25 Graph x Gridlines 15 Scale Legend Export Curves 5 Gr idlines Legend Copy Graph Biacore Word Pad Paint Export Curves txt Microsoft Excel Biacore A100 224 9
109. Sda 3 Biacore A100 5 77 Method 1 Overview Rm feray Congarialion unit Hill Fi as 4 Sampie L time as erered Soent correction Korreskan on kempese changes Soken Correction So corr LEme ssentensd Defene every SD cycles Ha sample lot Sampie Control Sample Zampa L Hm as entered Before every 21 cycles So 1 DD s 7 Bl x Optimize irc rm 3 Fusperwsaln 1 wlmt bu Boa cell 34 Lamrawer 1 2 General Settings gt 18 yQ PAZLA 5 ODORE ERE 1125 Method 5 Solution used For running buffer in Flow cell 1 1 5 Solution used For running buffer in Flow call 2 Overview HB5 EP Solution used For running buffer in Flow call
110. Biacore A100 4 73 gt f 174 14 2 1 5 4 4 Menu bar5RunoStop Run Biacore A100 74 5
111. eJVIC NUS S 8 E8815 N7 9 5 Contact time At least one high viscosity solution 35 40 Capture Commands L apture E Capture Insert Command Capture solution Capture Fel sp1 Remove From Cycle Capture Fe spot 0 Flow cell 2 Capture Fe3 spt Flowcel3 ENT TON DapueFc4spot 4 Down Contact time atl s a Lapture 1 Add ress spots separately por t Sample Capture solution Contact time C SUC S 5 Address spots separately spot1 spot5 Biacore A100 5 89 D Method Variables Evaluation Variables 2 Fc 1 Mame Fc2 Name Fc3 Mame 4 Contact Time s Diss Time s Flow Conc Numeric Molecular weight Numeric User defined variables Add lelete Li
112. ml Reagent B ml water available betore starting standby IF place all buffer uhinns ane bottle multiply the buffer volume bu 4 Desired standby time 14 EB 7 TV 4 1 Buffer 4 Status bar Instrument state Standby flaw remaining time 3 88 days Standby Flow 100 8 159 8 2 Shutdown 1 Desorb and Sanitize 16 70 Shutdown l File wiew Tools Run Instrument Help Ea Chip Chip Information Temp Rack Trav Temp Rack Tray Notebook Shutdown setup Run inina ne This procedure empties the flow system and shuts down the instrument The procedure iz divided into four steps Run time about 15 minutes Required salut
113. Biacore A100 Evaluation Open Assay Development Assay Conditions Surface activity test OK PET Notebook Close Settings bebe Next gt Tables Quality Control Save Continue Notebook Close Biacore A100 72 4 Evaluation Items Binding levels 1 Save And Finish
114. Evaluate Data Affinity Concentration Kinetics ResultPlot Sensargr am New ResultPlot j Biacore A100 190 9 Evaluation Items ResultPlot1 Evaluate Data Plot types Biacore A100 Report point versus variable x axis Variable Y axis Report point Response type 245 Absolute response Relative response Adjusted relative response Molecular Weight Adj Capture Surface Activity Adj Slope Flot types Y axis Paus Report point versus variable Report point binding late Variable Cycle number Report point versus report point Response type Fielative response j Report point for different curves Two repart points versus variable Ranking Report
115. Fedi d Dide Z3 Fez 3 Luce 43 Cure FET 3 _ E z0 Bampeongz rei Rii Wiem Berrya Zembik Hare he he ran pa poni enge Enel Extrapolate solvent Correction Extrapolation Extrapolation range RED ee o Extrapolation rangelx axis 100 176 9 Calculate i Hga Correction Setup b SeWentCoreclion HMolcuar Waekght P Surfaca Active Ad Basuhs Tha rureuz bakra wana cutrida cran unga Dsi nor apply Ki oue he mage keconmnende di F mbin coeden ki curear EE cce 1 E
116. P 148 O eae atan Dun aus 148 72 66 150 159551 y 153 5 oM po H 155 SB IOS ep 158 8 1 Standby Flow ee 158 RN NR Rm ECCE 159 SL e n Du un unu 162 SMS T 163 RN ON 170 I Fo RNN Ai i 171 DN T 173 EA NE RC 174 juo ec CN 179 dU uda eM er td ott 185 NN S s ND i 186 Mc 186 QUEUE 189 pn mE E 194 S BOR l u uuu O 199 EAE E deutet ies 201 AME 7 emi 215 c Y ku aan asas 217 ouo BE Es et 219 C 222 mE E E DQ y 5 O ibit 225 U enc Ei 224 Biacore A100 Biacore A100 1 Biacore 100 1 1 A100
117. Sample series 2 Base settings Connect to cycle type Sample l Biacore A100 6 121 Sample amp Assay Setup Startup 1 Select assau step n2 Select where ta define variable values for each Assay Step Define all variable values at run time series 1 Sample series 2 Define all variable values in method Define some variable values in method and others at run time 3 Enter values for Step Startup alphabetical order Flow Cell 1 Flow Cell 2 Flow Cell 3 Flow Cell 4 Name conc uM Name Name uM Conc uM Buffer amer Define all values in method Name Buffer Sample series 1 1 Select assay step 2 Select where ta define variable values For each Assay Step C Define all variable values at run time Sample series 1 Sample series 2 Define all variable values in method Define some variable values in method and others at run time 3 Enter values tor Assay Step Sample series 1 alphabetical order Flow Cell 1 Flow Cell 2 Flow Cell 3 Flow Cell 4 pami prakein x D buffer b
118. 120 Dissociation time 120 Biacore A100 118 6 BER Commands Capture Insert Command Hemove From Cycle Iove Down e Sample 1 k i r Regener 1 h i Cary over 1 Regeneration solutions Contact time Regeneration Regeneration solutions lmdMNaDBH Flow cell 1 lm Flow cell 2 lm Flow cell 3 lm Flow cell 4 Contact time s Atleast one high viscosity solution 10 mM NaOH 50 Compound 1 Create new cycle type or select an existing fram the list Sample Solvent correction Cycle types currently method Mew Rename Cycle types currently in method New Sample2 100 i 6 119 Sample2 1 Create new cycle type or select an existing fram the list Cycle types currently in method Mew Sample Solvent correction Delete Conditioning Samplez Hename 2 Insert the required commands 3 Canfigure settings For each command Commands Capture Sample Enhancement Regeneration Carp over Solvent Correction General Move Down Commands amp
119. 5 ml NI 2 3 Flow System Wash C O AAT AT S 20 011 23 22200 5ml 1 Biacore A100 T 147 7 2 4 Startup 159 8 2 Shutdown 12 Startup otartup Setup Run IF the instrument has been shut down Maintenance 1 Make sure the power to the instrument turned alf 2 the front panel and close the lewers the peristaltic pumps 3 Close the front panel 4 Switch the power to the instrument and walt until the instrument online Click Nest to continue Next gt Startup Setup Run Maintenance Place all reagent and buffer tubings in bottle containing about 500 ml deionized water Click Run to start the Startup procedure 500 ml Run gt Biacore A100 148 T
120. 7 by 52 1 5 4 20 2 1 Biacore A100 1 Biacore 100 3 t 7 1 1 44 DP FP A B
121. Azosulfamide Fe2 2 3 corr 0 2248 08 7 695e 3 2 804e 4 2 744 7 X 8 Accepted DK Carbonic anhydrase Azosulfamide Fc2 5 3 corr 0 4852 Du 7 581e 3 2 618 4 2 895e 7 2 9 Accepted OK Carbonic anhydrase Azosulfamide Fc3 3 or 0377 Oa 7831e 3 2 920e 4 2 682 7 5 kd 1 s ka 1 Ms KD M Rmax1 Chi RU 2 6 Biacore A100 206 9 2 Azosulfamide Response 100 0 100 200 300 400 500 00 au Time 5 kd 8 2b3e 3 1 s 1 45 77 AU 3 026 4 1 4 KD 2 732e 7 Validation info S amp nsorgram Table Parameters Conc 0 Initial Values Residuals Ente Validation message 1 Mote Data quality seems acceptable Validation info IO IU FY 3 VER Sensorgram Table Included Recalculate Modified Results Validation info Sensorgram T able Parameters Conc D Initial Values Residuals meeed
122. 0 Sample 1 Start lt gt CDbaseline 10 binding early 10 S binding late 4 stability early PCT 5 5 S stability late 5 Carry over CDco baseline 2 2080 6 2 co binding early LV PREGA DORIA 10 binding late 10 co_stability_early 5 5 co stability late 5 A100 12 Pre defined report points View Sensorgrams gt Wa ud 0 Ce 6 Jul xj Henr o Vae Cock 1 Zoom kek Adjusted sensorgram 30 HZA pH 0
123. 3 Buffer Fc4 HBS EP Solution used For running buffer in Flow call 4 General Settings Concentration unit Sample plate Analysis temperature after run uM 96 well Microplate 5 25 Cycle Types A Value 277 7 73 A 7 B Concentration unit Biacore A100 78 5 C Sample plate 198 Well Microplate well Micraplate g Deep Well Micraplate da4 Well Microplate Well Micrap D Analysis temperature after run Analysis temperature after run Biacore A100 5 79 3 Assay Steps Startup Sample Control sample BEREREHEEEREREREERERREEREERERRERERRERERRERRRRREREERRREREEEERRRERERREERERRERERRERREERERERRRERREREERRREREREERREREREEEERRERERERRERRERERREREREREERRREEREEEERRREREEEERREEREREERREREREREREREERREREREREERRREREREERERREERREEEREREREERERREEEEERREEREEEEEE x gt 1 Startup 27 7 7fIf 8318 0227 24 UE Rise 2 Sample 5
124. 3 PH scouting Setup Run 30 gimi HSA pH 4 0 30 ug ml HSA pH 4 5 30 ug ml HSA pH 5 0 30 ug ml HSA pH 5 5 15 ug ml HSA pH 4 0 15 pg ml HSA pH 4 5 15 ug ml HSA pH 5 0 15 ug ml HSA pH 5 5 60 ug ml pH 4 0 60 ug ml CA pH 4 5 60 ug ml pH 5 0 60 ug ml pH 5 5 Sample Fc4 30 ng ml pH 4 0 30 ug ml CA pH 4 5 30 ug ml CA pH 5 0 30 ug ml CA pH 5 5 NI pH scouting Metiad h yer eu Positioning Biacore A100 30 3 HSA pH 4 02 D Lagi 4 0 SO agir C pet 4 0 Spem pH 4 5 HJA pH 4 5 Ad pem CApHs s pii 45 Semi pH SO Apii 20 Land CA phi S 0 45 Haimi Hs 15 pH 5 5 0 CA pH 5 5 20 pH ss 45 SX oil Harc 80 mil Hace mb Ma iH 100000000 100000000 100000000 100000000 OOQOOOOG m m MC YC C MC 060 6 006009 IOOOOOOOO 00000006 Seri Run OOOQOOQO 00000099 Bottles S 7202 07 u U q 5 S SJ 0 5115 Tray 1 Bottles 64064 Estimated run duration 30mm Position Content Estimated consumption Water z100 ml excluding battle dead volume and standby consumption 390 mlidawi W ater Trays
125. 3 carr HS Warfarin 25 Series 7 Fc1 5 3 carr HSA Warfarin 25 Series 8 Biacore A100 gt Kinetics SN D F ts 103 3x108 FEBRE EM 107571 pU7U 272INWICXes7811 45 6 Affinity SIN 4 SD H 7 LA 0 10 Rmax Req 100 222 9 2 22 cave And Finish E sit Method Save Save As 0057 9 Fol
126. 3GHrHCIPH3 U Fol Gh Fel 3h HCL gH 10 HCI pH 2 31 FcVd 31M acl 2 1 21 3 Fz14 31M d Feii 318 5 Felt 3 iM Hati Fet 1 SON HCIpHZ C i Save And Finish Biacore A100 4 65 gt Evaluation Items Response levels Sensorgram Adjusted sensorgram el t 5 M o o o I I c E 500 P e m Z N 3 E amp p an i 3 m W 5 q 1 j t 350 500 q py NE QR n z m ls i ihi E E 1000 pe 4 10 B 11 180 Time Sample 1 Start
127. BIACOREA1OO mome User name c Password OK ie Ea Chip Chip Infarmatian Analysis Temp Rack Tray Temp Hack Tray Notebook Menu bar az ammin p macore ATIO Rn niri 1 m UM 2 Tool bar seachl data T3 hers Inkamalion es A Tre Stntus Method Builder Method NN E z Dbetrb cs 2 7 041 aiek L3 4 2044 2061 Method Eudder _ 1t 3 200d 10 20 e annt 7 3 11 Pagenaralion 50907 2053 Run Comphund 0 7 2005 11 51 T anh micro b niro interaction Bun Completed 1i 2004 5 48 er 1 0 4 5 7 3 7 041102 Feds iin Core ban Ten s smples 20tA Run Competed hre kinetik 4099 13240 m Conc Sabes bebzrz micea reni run 2 2065 1O 18 2005 11 54 2044 2061 5i Coni Seres runi 3 2066 Ruri 3 09 SCO Cor Series atasi mni z065 Pint head 1071972005 9 26 Fracedurez Conc Series beats mici 2066 bist hod Created 11 2 2004 2 51 31 PH m mesnhencemenck betaznicra 20015 JC 50830 2060 Run Competed DRINS 246 PT T HSA C smalimolecure rherarhon Run Compi amp ed 4e PT 1123220034 10 20 25 AM H PSA CA 25 2067 4
128. Compound x buffer buffer buffer 1 25 Com DOU nd A 200 Sample series 2 Compound x buffer buffer buffer Sample series 2 Compound x buffer buffer buffer 5 Sample series z Compound X buffer buffer buffer 10 Sample series z Compound X buffer buffer buffer 20 Y BRI U YI 11891228189 5 Positioning gt Trays gt StartRun 2 Biacore A100 124 6 6 3 gt 2815178 2 RET lem v Cla DMSO PBS HEPES HEPES Pe PBS DMSO 5 DMSO 10 DMSO DMSO
129. Linear 1 n D T E FclH 3 m RaM Fc 30 binding Jete Pos canol Fet 0 9040 riLnear zd Control curve x axis r Fel 3 RAM Fr 3D ui ibincirg 8 Fos cartral Fel Linear 0 8924 Fli Linear Carkral curve below x axis F 2 1 3 F a bwetazmicro 5uq ml binding late Pas central E Linear 41827 3 a betazmicro 5 late Pas control FeZ Linear 5618 Linear L i 2 Fra 3 0 ugil ibinding lebe Fos onvol LAH Linear Cortral curve below A m 3 r RaMTe3Dugtd ihndmglar E Fos oral Fet e _ m iCorinlcure bebe cmn _ F 3 1 3 l a betazmicro 5 binding late Pas control Fc3 Linear 01527 Linear E F 3 a betazmicro 5 ug ml binding late Pas control Linear ci 01417 Linear n n n ris L 4 3 RaM Fc 3nunm binring ete Fas cartral Fr3 0 8986 Linear zi Control curve z yz Fee 2 D ugil binding lete Fos contral Lines 1 9431 li Linear iCnrHrnl curve below x axis F Fedil 3 a betazmicro 5 binding late Pas control Linear 00271 F Linear E Fei 3 a betazmicro 5 bi
130. UV spectrometry PBS 10X 0 1 phosphate buffer 27 mM KCI 1 37 NaCI 1 x 1000 BR 1006 72 10 SAM DMSO 0 01 M phosphate buffer 2 7 mM 0 137 NaCl 5 DMSO pH 7 4 gt D DMSO Biacore A100 6 125 Solvent Correction WAA 222005 100RU
131. Y Change Buffers 1 4 146 7 2 1 Desorb 1 Desorb and Sanitize Standby Standby gt Menu bar Instrument Standby 4 Desired standby time E daps Make sure there at least ml Buffer 380 ml Reagent ml Reagent B ml water available betore starting standby NOTE IF place all buffer uhinns ane battle multiply the buffer volume bu 4 Desired standby time 14 Start Biacore A100 146 7 7 2 7 2 1 Change Buffers 1 4 10 40 ml 1 Standby Menu bar Instrument Stop Standby L Standby Change Buffers 1 4 7 2 2 Change Reagent A Reagent A 5
132. all tubings in warm water 40 50 Click Next ta continue Ip gut Next gt Biacore A100 144 T As admin p Biacore A100 Con 6M quanidew rydrochlori s or SDAOMSO 6M garisna hydrochloride or 50 DNSO sO dO mMHOolUNDNSO 7 Bottles 5aftware Run ew Rar E 100 el baitke deed volume ard standby coreumplion 35 volume and stendby E d Next gt Insert Trays Run gt Standby flow Biacore A100 T 145 10 Menu bar Instrument Stop Standby Stqndby flow
133. contra Fc3 pl Pos control Fed uM 66 Pos Fei uM iPos control 2 uM control Fc3 ym Pos control Fc4 uM H 66 Pos cortrol Fcl pM Pos control Fc2 uM Pos contral Fc3 pM PoscontrolFcd pM 66 Pos control Fc1 Pos control 2 uM Pos control Fc3 Pos control Fet 66 Pos control Fcl uM Poscontro Fc2 control Fc3 control Fc4 pM 66 Meg control ul Meg control Fc2 control Fc3 uM control Fes pf 66 control Fei pl control Fc2 M Neg control Fc3 uM control Fes ph 66 Meg control pM cortrol Fr M Neg control Fc3 uM control Fes uf 66 Meg control ul control Fc2 control Fc3 uM control Fc ph 66 Neg control MM conkrol Fc2 NegcontrolFc3 control Fes M 66 Meg control Fct nM MegcontrolFc2 Negcontrol Fc3 uM control Fc pM 66 control Fei ud Negcontrol Fc2 control Fc3 uM Neg control Fes ym 66 Neg control MM cortrol Fc2 control Fc3 uM control Fc um 66 Meg control ul Meg control Fc2 control Fc3 uM control Fc pi 66 Neg control Fel uid iNeg control Fc2 W Neg control Fc3 control Fes um 66 Regenerabon Fc Regeneration 2 Regeneration Fc3 Regeneration Fc 1611
134. point versus report point Plot types amp Mass Report point versus variable Report paint binding late Report point stability early Report point versus report point Response type Relative response Response type Relative response Report point far different curves Two repart points versus variable Report point for different curves X axis curve Flat Axes Report point versus variable Report point binding late Report paint versus report point Response type Relative response Report point far different curves axis curve Fe 3 eorr C Two repart points versus variable Two report points versus variable x axis Variable 2 Plot types Left y axis X axis Report point wersus variable Report point binding late Variable Cycle number Report point versus report point Response type Relative response Sort Report point for different curves Right y axis Two report points versus variable Hec E IE Response tupe Relative response z Biacore A100
135. some variable values in method and others at run time alphabetical order Assay Steps j Regeneration 1 Cycle Types General Settings 3 Enter values for SteNReg condition 1 buffer buffer buffer mouseIgG 2ugml buffer buffer buffer 10mM gly HCl pH2 5 Sample amp Assay E EE Setup 3 Enter values far amp ssay Step Reg condition 1 Sarin alphabetical ar J F Regeneration 1 Flow Cell 1 Flow Cell 2 Flow Cell 3 Flow Cell 4 Flow Cell 1 Flow Cell Flow Cell 3 Flow Cell 4 buffer l s buffer buffer buffer Verification Setup A100 4 61 patirs Setup Run ETT Bhi illie zh E A E times as Seling bor step en condim 1 o 4 I E T i 7 E S BER D Sample 1 120 Optimize ka Loss Sample Conzumpsion E 4 T Sclbora 15 Dro Es in Lis FT Fun l EJ v A ww ES v Berni Hus Biacore A100 62 4 JH amp RTEFRHBIGEODIS600D2E T IRS Cycle Run List
136. test buffer buffer buffer Proterin 2 Juernyiey Ligand activity test buffer buffer buffer Proterin 2 ig Ligand activity test buffer buffer buffer Proterin 2 10 Ligand activity test buffer buffer buffer Proterin 2 Semple amp esey Ir did test buffer buffer buffer 2 Setup Ligand activity test buffer buffer buffer Proterin 2 Ligand activity test buffer buffer buffer Proterin 2 14 Ligand activity test buffer buffer buffer ProterinA 2 28 Ligand activity test buffer buffer buffer Proterin 2 Cycle Run List Ligand activity test buffer buffer buffer Proterin 2 Ligand activity test buffer buffer buffer ProterinA 2 Ligand activity test buffer buffer buffer Proterin 2 Positioning 19 SN test buffer buffer buffer A aaa 2 20 Ligand activity test buffer buffer buffer ProterinA 2 25 Ligand activity test buffer buffer buffer Proterin 2 22 Ligand activity test buffer buffer buffer Proterin 2 Ligand activity test buffer buffer buffer Proterin 2 24 Ligand activity test buffer buffer buffer Proterin 2 25 Ligand activity test buffer buffer buffer ProterinA 2 i Positioning Trays Start Run Biacore A100 4 71
137. within Sample Every El cycle C Distribute occurrences evenly Run assay step once first Run assay step once last 2094 22 JU Cycle Types Biacore A100 6 133 Cycle types currently in method Sol corr Method 8 Biacore A100 Biacor Maintenance Kit type 2 BR 1006 51 95 ml x 2 BIAdesorb solution 1 95 ml x 2 BIAdesorb solution 2 65 ml BIAtest solution in HBS N 10 ml x 3 BIAdisinfectant solution conc 90 ml BlAnormalizing solution 50 ml HBS N buffer 10X 1 Sensor Chip Maintenance 100 T 135 Maintenance D elector Preparation B Hydrodynamic Addressing otartup G Fon System Preparation Change Buflers 1 4 3 Change Reagent Startup Published Fracedures WI Flow Sys
138. 0 5 95 Method Sample amp Assay Setup Define all variable values in method setup Run 2 2504 UL JU Ml No i Biacore A100 96 5 5009 Setup Run t sete Buffer Buffer A35 472 A des ATE AFF Los ATO An Padina AB Ani is Ez EX BE B3 15 EEJ 83 Bra Biacore A100 5 97 Positioning 97 23 o p iO Setup Run Eontentrez tontentres Content Fea R14 iSpgi pH 4 0 etp CA pH 4 0 REI 45 Spem SA pH 4 5 engl CA pris pet 4 5 45 Rlar 0 S D 1 pH SO CA phi 5 D 20 50 45 P HH BB 31 pH 5 5 15 5 5 EnualmlCanH5 5 mmnmlCapHS5 45 l mM MdH 150 mil Hac mM
139. 00 168 9 ITables RU 170 1 1 171 12 173 1 3 de DC Approve Data Report Points Kayword Tabla mw ses ww Point krd Signal Correction Signal Correction 2812 2472 174 14 Quality Control Biacore A100 9 169 Quality Control 179 1 5 Descriptor Based Quality Control Bascire Quaity db Amzociafion Quality 4k Dissociation Quality 4 Summary Tabla A NES m 3 Buck H si gt
140. 10 10 10 10 A53 A65 A77 489 10 10 10 10 A54 A66 A78 490 10 10 10 10 55 6 A79 491 10 10 10 10 A56 A68 480 492 10 10 10 10 Control sample Negative Negative Negative Negative Control sample Positive Positive Positive Positive A57 A69 A81 493 10 10 10 10 58 A70 A82 A94 10 10 10 10 Positioning Trays StartRun Biacore A100 6 115 6 2 Se lt RIEDI gt 1 1 2 3 t e C 2 Fc1 2 2 FC4 Compound x EA foe 2 foe 14ff 285 1 5 10mM NaOH 30 gt Startup 3 Sqmple series 1 protein X Sample series 2 Compound Biacore A100 116 6 lt gt New Method Biacore Methods Kinetics and affinity Kinetics and affinity OK
141. 15 Sample sulfanilam aE E Fcz3 3xorr 2 1 Rahhitzerum alb Corrected 9 16 Control sa 1 Meg contri z Wi1 grnrF F I Donkey serum alb Corrected 17 Control Meg contro 1 Meg contri v Frl 4zxor 1 5 Porcine yerum all Corrected Dente Feit Axor a Human senam alb Corrected I a E edd 3m re 4 1 Rablil eium alb Corrected 1 d 3 1 ZF mir i iab L Lid x M arah 2 g zz Sample 50 IM Na GERE e Fcz 5 4dzurr 2 05 Human serum alb Corrected sa cantra 1 Meg cente gt Feds 4xmr 3 Porcine seum alls Porrected e m le Fez Amxarr 02 2 Rat seram alli Enrreciedi k r Vic za ranas ea ud Inoki Selected Exchude Al Erdue Selected EEE Inckade Selected KHEN i adjustment Adjust to injection event uc E Zero at injection position Sample start Y adjustment Adjust to report paint b 1 zero at repart paint position baseline X Y No adjustment Adjust to report point dude ot Adjust to injection event Te
142. 27 2 2 amp 1 20600 5 3 i Z j 00 __ eo 5 inn E enin P 5 Fi i E 2000 n i L 2000 V O IE E c vA ps gt lt o gt s gt pO h 9 SaNs p 9 9 gt gt gt lt amp gt gt s v gt y D O sN n 5 100 50 200 50 300 n d Start coe lt gt Report Points User defined report points Add c m Weeis Pos Relative Ta Winden s taal Relative Report Pom Ja www n e ii A iii srl Sample E idd 5 Name 30 Time RPO Position Position BeforeStart BeforeStart AfterStart BeforeEnd AfterEnd BeforeDissociationEnd AfterDissociationEnd Biacore A100 172 9 Relative To Relative Tn Regeneration Regeneration Sam
143. 8 1 4 20 Fc4 5 3 103 52 5 9598 168 0 29444 0077734 0286238 Ok 18 1 B 20 Fc45 3 103 158 5 9507 32 0219592 0 067969 0 200162 Ok 20 1 B 20 45 3 103 167 5 9597458 0344315 0 015569 0383576 21 1 4 20 Fc45 3 103 757 5 9596316 0197841 007037 0165114 22 1 3 11 IFcd1 3 13 32 10 1264438 0 330736 0 02157 0340371 23 1 3 11 31 3 13 52 5 1264852 0 314187 0 06641 0 322643 24 1 3 LER Le 3 13 158 5 12644 83 0 329743 0 039174 0358443 Ok Biacore A100 186 9 9 2 Approve Data 9 2 1 Screening Screening with capture Evaluation Items Ranking binding levels Evaluate Data Settings 4 Plot de Result Thumbrai ratis Solby cave Dee se a RET MEL LEM i mate Pa wu Ez1 1 3 800
144. 89 Stabilization period after injection Flow rate 10 30ul min 100 5 91 Variable Select assay step Select where to define variable values for each Assay Step NN Rg 2 Select where define wanabe values Tor each Assay Sep Method 1 Define variable vales al run fme 1 Defina sll variable valves in method B Dehne iome wanabe method and at ime m n n 3 Enter values Tor P kay Harup Sorlin alphabetical order Buffer Buffer Actions Define all variable values at run time Setup Run Sample amp Assay Setup Define all variable values in method Define some variable values in method and others at run time
145. Control sample Sample 5 Control sample x Assay step Negative _ N Biacore A100 80 5 Assay Steps 5 Method Th by Zampa as entered A buon In Sample vi ri mones Ihi iaga Sapia 1 tire ai entend Haw Sliep oben coretan t Soent Correction 5o com tiga 35 Paene f ivir SI cus Hemos Step Control garrilE __ Me Contro sample BE 1 tie 35 Before gvern 20 Setup Fur B Repeat asse chep T ce f zi norunt moins step onze Irak Rurasepzenonselas 1 2 11 21 kas 1 1 2 2 33 Random A B D A Assay step Startup Startup Sample 3 times as entered I Sample Sample Sample 1 time as entered Solvent correction t Solvent Correction Sol corr 1 time as en
146. GE Healthcare 3 Biacore Biacore A100 Ver 1 1 Instrument Handbook Biacore A100 1 2 3 Biacoe A100 08 ccm 1 7 6 5 RU p 6 J OO 6 2 u tinte 6 qc 7 7 P RE NAVEM m TT 8 E 9 9 Sm utu 10 mos gt LARUM ADAM DEA 17 T P MER ESTILO E RM 18 inus Ape SS SEES eut 19 impete qu MEI ML MR 20 20 21 22 oe 23 RE EN 25 3 2 U 237 pH 1 4 26 sp UJ 7 OU i 27 D e 32 2 0000 34 T RR NES 36 c c s BINE ET S edet ete eerte ttl ette deett 38 2202900 RS esse ln ae n iui 39 c r a E a 40 3 3 3 2779 34 8 tritt ttti 40 7 41 s np Sas n s 42 100
147. Menu V 7 5 Automatic Positioning 22 51 Automatic Positioning E x Change the order in which the samples are positioned by ordering the regions ControlSample LimeGreen E BottomLeft 27 ID Auto v lv SolventCorrection LC Gold lt Column de BottomLeft m Reagent Auto E RegenerationReagent E Tomato Row dz BottomLeft Reagent lt Auto El Pooling Auto Region Pooling Ves OK i Biacore A100 5 101 81 1 2 2 Tray 3 Bottes Automatic Positioning Color Orientation Anchor Plate L bh v4202Z7v Separate tray Start Row 1
148. System Result run ID System Check 2006 01 11 15 13 37 User ID administratar Run Date 11 2006 15 13 software ID Biacare 100 Control Software ersian 1 0 Instrument ID 20102 Configuration ID 1 4 Analyze Temp 25 0 C Flow cell leakage test OK B Refractometer test OK C Reagent pump test OK D Syringe pumps test OK E Hydrodynamic addressing test OK OK BAD BAD Biacore A100 158 8 AIl 8 Stqndby flow Shutdown Shutdown 8 1 Standby Flow Standby flow 4 Menu bar Instrument Standby gt Desired standby time E daps Make sure there at least zuu ml Buffer 380 ml Reagent
149. a bes fo Mag iananaa Chen Ireinamant 20102 online Analysis lempore 25 0 25 D C Tr taraparalurs 13 0 15 1 heisirument He temor chip inserted idle Sensor chip Chip type Chip ID Chip lot number 0 9 Methods Normalize 10 Hydrodynamic Addressing Normalize 20 Normalize Hydrodynamic Addressing Next gt Biacore A100 12 2 admin 2 100 Control Software Run New Run Fie Took Instrument jt G z ch Chip Chiplnformmsion Anaysis Temp Rack Tray Temp sk Notebook Setup Run Detector preparation ee oO Status Instrument 20102 online Analysis temperature 25 0 25 0 Tra
150. aliee before Hydrodynamic addressing Do want ta normalize Don t normalize Biacore A100 7 X 151 Ji V Normalize Next gt viral 5afbeare Rain Pes Han BlAnormalizing solution Next gt amp Insert Trays Run gt Biacore A100 ais j Docking EE enne tr ai iridis Biacore A100 Hydrodynamic addressing Result run ID Hydrodynamic Addressing 2006 01 18 14 30 31 User 10 acrminisiratar Run age 2006 14 30 Software Eizcore Control Sofware Worsion 1 la i ID 20102 CenfiqurationlDz IFCIDA Analyze Temp 25070 Respense Drit 4 2000 AB 174 217B 14 21378 s 2 1 21 7 Li 2180 2102 Z168 22222223 corr fota A100 T 153 7 3 3 Spot Configuration Hydrodynamic Addressing Bad DYZ YTRET d 150 mM 8105 HBS
151. br zeil 1 ml II En ul 11 zampim FE n T zampiml EN n AT ER DG ES l En Serpil mA n 4 ji HHL EE danki PA En 9 Pen m Ak TTA zergal EA n Aes EDD zeil En Ej AEMILIA 10015 EE rr TEL LE mT Pc2 3 n qus l PE n mm e UAE Lr e oed En Hb Sample cycles 88689400 77 9 Dine f Injection Do not qpply solvent correction to curves outside the range recommended lcg woe ATL Calculate Biacore A100 Weight Adj 100 RU Da 100 G Capture Adj
152. capture Concentration Concentration OK i Biacore A100 166 9 Settings 25 47h v 0512 Approve Data SpotSefngs 49 Seket Cumes M s G actwe gpi ming pa all rpaiz in call Carbonic anhydrase 1 Amine coupling T rana I cwn T 4T RU 2316AU Tm RU Hi ruina coupling I rari 10 Dura cepi 1D Janina ceapling 10 ruri uciradim i Er Ri Fj THH Pii Om 3 6 3 ETT DST 84 ping risi capile 1L nini E STI pr RU nE Fij jait ABR I 1 Fi i GIA GET GEL a 10 IT oi ples 10 nii LLL rg LE LT T 4581 13 Hu Umes HAJ Active J Reference Feterence HERES Undefined Edit mode
153. ders Search Small malecure 3 items C iae CA Azo kinetics Run Completed 2004 05 25 22 17 Import user e kinetics Approved Data 2007 03 13 11 02 administrator m G ss G G EA CA Azo kinetics Evaluation fUnfnished 2007 03 13 15 36 administrator 71 DNA DNA immobilization 1 instruction J liposome Ca manual Ca serum e Small molecure Ca Test Type Evaqluation Stqtus Unfinished Save And Finish gcc ce 3 Kinetics and Affinity Kinetics Affinity Folders Search Small malecure 4 items J a Assay Modified Modified By Folder C Mii Azn kinetics Run Completed 2004 05 25 22 17 Import user 2 ERES kinetics Approved Data Finished 2007 03 13 11 02 administrator T L dL Azo kinetics a aa ac NANNERL 2007 03 13 15 42 administrator x E immobilization 1 C Azo kinetics finished ZEvaluation 22007 03 13 15 44 administrator Instruction 71 liposome 21 manual
154. ession raabe 20 865 x 0 996 0 996 1 000 0 998 FCI 1 000 0 997 0 998 1 000 Difference preziaus ri amp w u 2 RERE EEE of max rolative response Y iriet 50 2 53 2 5 32 554 38 596 36 395 66 15 51 2 e 18 OK Hydrodynamic Addressing l Normalize Setup Run Biacore A100 T 155 7 4 Desorb and Sanitize pep Jam Al Td 5 0 1 BIAtest solution 2 HBS N HBS N Series S Sensor chip CM5 System Check oystem Check Setup Run Select tests to run Click Hest ta continue Leakage test B Refractometer perfarmance test Reagent pumps test vi vi vi kel D Syringe pumps test vi vi vi E Hydrodynamic addressing tes
155. ians Deionized water about 200 ethanol about 100 ml You will also need the to open the front panel Click Nest to continue Next gt AW File view Tools Run Instrument Help i Ea E qm Chip Chip Informatian Analysis Temp Rack Trav Temp Rack Tray Shutdown Setup Run ee This step flushes the whole fowm system with water and then empties the af liquid Flacg all tubings in deionized water Click Run tn start Step 1 Run gt Biacore A100 160 8 2 File View Tools Run Instrument Help Analysis Temp Rack Tray Temp Rack Tray Ea Chip Chip Information Shutdown D This step flushes the pumps with 0 ethanol all tubings In U ethanol Click Run In start step 2 70 Run gt 3 File wiew Tools Run Instrument Help E Je ER Chip Chip Information Analysis Temp Rack Trap Temp Rack Tray Matebnok Shutdown Run ones Step 3 This step empties the pumps and pump tubings af liquid Let the tubings hang in the arr Click Hun ta start
156. low cell 1 Buffer Fc2 HES EP 4 Solution used for running buffer in flow cell 2 Overview BufferFc3 HBS EP 72 7 Solution used for running buffer in flow cell 3 BufferFc4 85 2 Solution used for running buffer in Flow cell 4 General Settings Concentration unit Sample plate Analysis temperature after run Assay Steps uM 36 Wel Microplate 5 Cycle Types Biacore A100 48 4 Assqy Steps Method Fair or nal Sarpa Sample 1 as entered sese eet ete we Fotie Negative control 5 Remove Step Cycle Types Capue gt Regeneration 1 i Darry aver 1 Commands Regeneration1 Carry over 1 Remove From Cycle A100 4 49 Commands Inse
157. nding late Pas control 4 d amp UIb5 Linear Fet 3 i RaM Fc 3D lote k Fos onvol Fet Linear Linear Corkral curve x axis 3 5 3 RaM Fc 30 ua binding lete Fos Fet s Linear Ds kli Linear Corkral curve below Mies amp opustme nt 50 1 66 50 D 8 2 Biacore A100 178 9 Batch select curvetype for all rows Active ReferenceSubtracted hi Corrected IET 9 Batch select reportpoint for all rows m binding early binding late stability early stability late
158. nds Capture Sample Insert Command LowSampleCensumpion Remove From Cycle Flow rate ln A Contact time E s Down Dissociation time NE s pU Sample 1 lt Regeneration 1 Type Low Sample Consumption Flow rate 30ul min Contact time 120 5 l Biacore A100 60 4 OD TS 60 OD2 HS SJ Commands Regeneration1 Commands L apture Regeneration Insert Command Regeneration solutions Flow cell 1 Hgn Remove From Cycle eg Flow cell 2 Regi Flow cell 3 Regi Flow cell 4 Move Down Contact time 1 z s At least one high viscosity solution Regeneration O Contact time C F3 57878 OON DOR fe e ALCUN 7 d 5 8814 60 Sample amp Assay Setup buffer 1 Select assay step 2 Select where to define variable values for each amp ssay Step Method Reg condition 1 Reg condition Define all variable values at run time Reg condition 3 Overview Reg condition 4 Define all variable values in method Define
159. on Quality 4 Disseciabon 4 Summary Table Seleci lon pl carie 10 Fc1 1 3 sc4 1 K ES 1 Flow cel celi 2 Flo celi 3 cald an ep ee E 18 Sample n 2 16 Sample Furosemide iu 2 i i c DE E NEN T U ane 0 12 CrrtroEarWpl ian r 2 i T 2 AM M HO dank PME i 4g ms iu 2 5 e WP EE o i Contro iuri int ki Respo A 50 50 100 150 250 300 Al Include Selected Esclude Al Selected Time lt Back Select flow cell and cycle 20 6 71210 WM D SN Select curves Select flow cell and cycle Excluded Sensorgrqams Biacore A100 182 9 B Plot based QC
160. ple Window 5 5 Baseline 0 Relative report point Relative Report Point baseline ODE 10 4 C Regeneration amp U baseline T Regeneration B 151 xl 1 T m Sierra Fe1 Adjusted sensorgram 30 HSA pH 4 0 3 rame t gnmm E o 5 e E r4 s 5 N E ii N 1 25 im Hi f E ne Ls zcna OTHER nm ni 1 M u zu 150 E 250 300 d Start Hep Close Biacore A100 1 3 Report Points Keywords
161. pose General Evaluation variables Sample amp Assay Setup Biacore A100 10 6 Sample amp Assay Setup Method Variable Startup Define all variable values in method Name Buffer Biacore A100 6 111 Control Sample 5 control Fc2 Define all variable values in method Conc uM MW Da ible values at run time le valjas i h melha 4 able values in method and others at run lire p ER Name Sample Flow Cell 4 Name Sample i BS TE i sur Sample iFlow Cel1 MW Sample i Flow Cell z MW i Sample FowCel3 MW Sample Flow Cel MW i Define all variable values at run time Verification Biacore A100 112 6 Verification fotom
162. ption 72 100 excluding botte dead valume and standby consumption 72 mliday zt ml excluding battle dead and standis coneomplion 72 Bottles amp Next gt Insert Trays Run gt Biacore A100 150 T 7 3 2 Hydrodynamic Addressing e 2L1I EJUO AT RF Sot 89 3SEIRNTLU CL 0 Dock Normqlize 20 BIAnormalizing solution 150 mM NaCl HBS TESTES Hydrodynamic Addressing Hydrodynamic Addressing Setup Run Use suitable buffer recommended buffer 5 in battles 1 4 Maintenance Use distilled water in the water battle and buffer or water in bottles and B Click Next ko continue Next gt Hydrodynamic Addressing Setup Run ee It iz recommended to run Morm
163. qp WE G o X adjustment Y adjustment Biacore A100 196 9 Zero at injection report point positio Group cycles by Group cycles bu m Legend and Flow cell Curve Single cycle Legend tooltip Legend and tooltip Cucle Curve Name Cycle Fc Ligand Cycle Sample C Cycle Ligand Sample Biacore A100 9 197 FY Data selection Sensorgram 1 Eyaluate Data Settings 4 Sensorgram udiuct bo injector Zein at position 22 E Cycle Curve Hame C Fo Ligand C Cycle Fo Sarple Fe Ligand Adjust bo HOJ Appreed Dasa lere Ei Z Eria Heres LET SL TI Hace Bufer Bale paj
164. re is at least series that should be inspected Do vau really want to proceed Yes Yes i Finishing and lacking affinity evaluation Affinity Save Save As Kinetics Affinity Save And Finish 221 22 Biacore A100 19 Evaluate Data Settings 4b Preparation Examination Results rate FL Apporasa Henn 2 Cabaret Fel Aca Ph d And Finish Fel ll Carbone Ps 21 DHL Sai Ranges Azosulfamide Salby Smis Diesin al Fel 2 ankpadeasa bacini Fd s ues rp Se
165. rint Report Point Table LMS Data Rum evaluation Evaluation Trail Import k ExiE p Menu File gt Export gt Report Point Table TERLI PIILEE DD TURES S Microsoft Excel D F H I K L 1 lFc v 8pot v Curve Ne v DiodeRo v T ime v Window AbsResrc v 5D v LRSD v G uality 1 3 15 35 3 45 32 10 10042 06 0272364 00432 0244163 3 1 3 15 35 3 45 52 5 1004091 0398818 0 082143 0411461 4 1 3 15 Fc35 3 45 158 5 10042 010712 002813 010432 Ok 5 1 3 15 Fc35 3 45 167 5 10042 45 053064 0 25619 0 254594 Ok 6 1 3 15JFcd5 3 45 757 5 10041 57 0213477 0 017355 0235898 7 1 2 10 25 3 197 32 10 1094087 0321033 0 064702 025169 8 1 2 10 Fc25 3 197 52 5 10937 96 0195167 0 036719 020424 9 1 2 101 25 3 197 158 5 10938 83 0196332 0 035379 0206656 10 1 2 1OIFE25 3 187 167 5 10939 92 0491286 0162835 0 430927 Ok 11 1 2 TOIFC2 S 3 187 757 5 10939 26 0 222532 0 06607 0 206886 Ok 12 1 1 312 172 32 10 4329 05 0314064 0 057457 0 263146 13 1 1 HESS 172 52 5 433047 0196957 0 04487 0199206 14 1 1 172 158 5 4326 928 013249 0 011384 0146203 Ok 15 1 1 172 167 5 4326039 0178552 0 08711 0 081568 Ok 16 1 1 1 JFc11 3 172 757 5 4322 549 0192608 0 007199 0214815 17 1 B 20 Fc4 5 3 103 32 10 9598 023 0 512543 0126634 0 309667 Ok 1
166. rscezrem tale Fel ch Dit odd in Spi urs E 15 2 GHL 4 Furosemide3 Response Help 5 to injection start 3 0 s 1 Biacore A100 216 9 Advanced xl Amas Global Replicate measurements coincide Local Offset during injection Al Initial values Local ka anners Ms Constant D kd 1 00063 1 5 kIg Help Cancel Rmax Global Local Offset during injection RI Constant 0IRUU Local Initial values Biacore A100 20
167. rt Command Remove From Cycle Move Move Down Sample 1 rate 6 Contact time 32 E 3 JUS time a s CAU Normal Flow rate 10ul min Contact time 120 s Dissociation time 120 s Sample amp Assay Setup i Z Select where ko define values ior each Assay Sep Deina all yaishie vouer at run lima C Defra al yaishle values method eannbhe vezes nasa and ak nun bang General Sgtings Asse Sieps Types Al Biacore A100 50 4 JH amp TEFHRIGE OD 7260002 T HS l Verification Setup Run Sample amp Assay Setup Setup Run Positive contro 2 buffer List Biacore A100 4 51 Cclc Assay step name Sample Sample Fcz sample Fc3 Sample Fc4 Conc Conc Fez Conc Fea corral buffer b
168. step 3 Run gt i 4 File wiew Tools Run Instrument Help gt Chip Chip Information Analysis Temp Rack Tray Temp Rack Tray Notebook Shutdown eee os Turn aff the power to the instrument Far the connection to the instrument to closed Click to continue RENERE ELT Next gt Biacore A100 8 161 i La 9 The connection ta Biacore been lost i pes dus e pee cm eS cC S Biacore A100 162 9 9 Biacore A100 Evaqluation 1 Biacore A100 Control Software Type Run lt La
169. t F test G Buffer selector test Next gt Biacore A100 156 f Setup Run IE HIIHLIE Next gt Setup Run 96 i Biacore A100 Bottles iim m BHiaesre A100 Coniral 5efiware ium mn Setup Run Beatles Tus gd Belles 143184414 Tu adim A E 1 z 3 4 sition Content x Estimated consumption LET 5 100 ml Bh rary bot ded me and cenguamption 1390 mijday p m aa n innn aaan koi a LLL LLL m m ve LLC consumpti 72 mida zii ims SER TUER AD 172 andare zU ml botha dead volume and oonsampeien 72 THari ml exdudng battle dead wolume and sadir consumption 72 mida 71277 3 Y B 2 HBS N Next gt Insert Trays Run gt 100
170. t the procedure after pressing Run 90 ml BlAdesrob Solution 1 Run gt Biacore A100 Desorb and Sanitize FI ME 90ml BlAdesrob Solution Run gt Desorb 5anitize Biacore A100 7 14 1 Desorb and Sanitize Run lt Memes Step 3 5anitize all tubings with a clean tsue Insert all tubings in minimum 105 ml diluted Bl amp disinfectant Solution diluted according to Instructions Far Ll se Click Run ta start the procedure NOTE Do not abort the procedure after pressing Run 105 ml BlAdisinfectant solution ZAH Run gt 6 Desorb and Sanitize Hun Meienance Wipe all tubings with a clean tissue and let them hang in air Click Run ta cantinue Run gt MS Desorb and Sanitize Run Insert all tubinigs water Click Run to start the procedure NOTE Do not abort the procedure after pressing Aun Run gt
171. tem wash Spot Configuration Superclean 3 l est System Check Detector Preparation Hydrodynamic Addressing s Hydrodynamic Addressing I i Normalize This procedure calibrates the flow system for correct addressing of detection spots within the Flow cells pini Total Run time is about 24 minutes without and 35 minutes with normalize included an Change Reagent Last run 2006 03 24 20 43 56 l Shutdown w Startup Biacore A100 136 T 7 1 7 1 1 Desorb 20 1 20 20 Toolbar Rack Tray Temp Je PUYU Rack Tran Temp Set Temperature Hack Iran temperature 20 20C OK cce d es
172. tep Simulate Cycle Hun List 6 Remove Step Reg condition1 EREI SPAART Y ERRU Bose setting amp 589 6 gt x New Step times as entered Assay step 1 Sample Nok connecked 1 time as entered x Base settings Number of replicates Base settings Mame RHeg candition 7 Purpose ambe T Connect to cycle type Sampe Number of replicates Hz times As entered 1 2 3 1 2 3 Drder 1 1 22 33 Random Cycle Types Biacore A100 4 59 1 realg a new cycle type pr select an exesting Fom e list Method Tha in in rampis Poudi ef and 2 ihsan the required comanda Sens 3 Congue settings for ach command Cycle Types spine insert Command Semple amp As Venticatian Commands Sample1 Sample f C 37 0 0 7 0 sS ES 5 Comma
173. tered Before every 50 cycles Control sample t Control Sample Sample 1 time as entered Before every Z cycles Remove Step Mem MoveDom Step Pea Oj Simulate Cycle Run List F LO 7 1 22 Biacore A100 5 81 Base settings Base settings TE Startup Furpase startup El Connect to cycle type Sample Name Purpose Purpose 1 Sample 2 Calibration 3 Conditioning 4 Control Sample Sample 5 Undefined
174. uffer bufer masalga Setup Run Peste Positioning Trays Start Run Biacore A100 52 4 1 2 A100 Evaluation TL L Ege eb Open i Assay Development Assay Conditions Binding conditions OK Pers i Close i Settings Next gt Tables Quality Control Save And Continue BSE Notebook Close Biacore A100 4 53
175. uffer i buffer es ss y 522227 ri E Soc Be 71 777788 71770070707 WT p f x WO S 7 E mim E S s ss NE 25 bue 5502 77 i e ss s 13 buffer pu mi OD GS xc TREE EE e Define all variable values method 25 20 10 60 Name c Conc uM Flow Cell2 4 buffer Conc uM Biacore A100 122 6 Sample series 2 1 Select assav step 2 Select where ta define variable values For each Assay Step Startup Sample series 1 Sample series z Define all variable values at run time Define all variable values in method Define some variable values method and others at run time 3 Enter values for Assay Step Sample series 2 Sort in alphabetical order w Flow Cell 1 Flow Cell 2 Flow Cell 3 Flow Cell 4 opem eem meme Name Come name Compound x 0 buffer i buffer i buffer ERE a 07 ee n ME HH MEE 545 buffer AS MEME T ME ELE a T TEN T EE E n Mo M EE 2 buffer s
176. w cell 1 Flow cell 2 Flow cell 3 Flow c 1 1 1961 buffer buffer buffer 1 E 1 2 rh I iga buffer buffer buffer Ligand overview Spot 2 Flow cell 3 Flaw 4 nh ln 1 2 rh l iga 3 Unmodified 4 5 Run Order Chemistry Amine coupling 10 min activation Flow cells 1 5 pats 1 3 Ligand injection Contact time min Spot Flow cell 1 Flow cell 2 Flow cell 3 Flow cell 4 1 1 1 anti IgG butter buffer buffer 10 2 hi I a buffer buffer buffer T r B Positionin SETEN 812289189 S Biacore A100 40 3 3 3 2 Result Immobilization result Run Flow cell Spot Chemistry Usage Ligand name Contact time min Level RU Fc1 4 Amine coupling 10 min activation Immobilized 1962 1 23411 n Sensorgrams 5 Amine coupling 10 min activation Immobilized 1561 10 45492 by Flow Cell ns all Sensorgrams Run Information Sensorgrams by Flow Cell All Sensorgrams 3 3 3
177. y temperature 15 0 15 0 Instrument state No sentor chip inserted idle BlAnormalizing solution BIAmaintenance Kit 380ul F DREZ LL wg 18 4 i Bottles ray t1 Bottles ig a 8 8 Water 1 2 3 4 Estimated run duration 37mgn 4 Position Content Estimated consumption Water 100 ml ni bottle dead volume and standby m en 390 mdawi Next gt Biacore A100 2 13 admin 2 Biacore 100 Control Software Run New Run EERE View Tools Insbrumwnt Help EJ Eh E c m Lhiplrnfcrmson Anaysis Temp Rack Trey Temp Mobok Setup Run Detector preparmiom 17 f Resgert plote 95 Well Mioroplste z Trays Emphy Storage Urat Inslrurment 207102 temperatura 25 0 25 0 E Tray taenperalure 151 qaem c Instrument state Mo santor chip Insert Trays Preparing For rack trap handling 0 30 2397 EL ne I2 ES amp Biacore A100 14 2 Insert Tray

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