Biacore 2000 （Ver.3） 取扱説明書
Contents
1. 36 36 A ESO TIA Es CES O JC Cycles2. 1 1 Laongmuir with mass transfer Run Run Application Wizard Kinetic Analysis S3 Kinetics Analysis Steps EE Kinetic Analysis Steps Concentration Series Control Experiments Analyses the binding of one analyte concentration at a range of fow rates Results are presented as an overlay plot aligned to the time of injection Help 4 Back Hest gt Cloze Mass Transfer Next gt S3 Kinetics Assay Principle EE Assay Principle Binding Using Capturing Molecule Eaptuning Parameters Hame Ligand Elow Rate E uz min Injection Time E min Stabilization Time 10 min Help lt Back ext gt Cloze Direct Binding
3. R Bulk Effect DE PIZZA ARO RA CONIL ERIN 30r ng ml BE ug ml EIREEEAS ADS SAGA PRESOS Kp BRE RIZO 1 10 10
4. 7 Keyword Es Keyword Cond vr Cancel A Fc APROG Time bsResp Slope Baseline RelResp Id 3 screen 2025 238470 0 06 Yes 0 base 4 screen 2025 32276 5 0 03 Yes 0 base 4 3 screen 2025 84295 0 09 Yes 0 base 3 screen 3225 242235 0 00 No 376 5 bulk Furosemide 4 screen 3225 326775 0 10 No 401 0 bulk Furosemide 4 3 screen 3225 84540 0 11 No 245 bulk Furosemide 3 screen 352 5 23837 3 0 27 No 9 8 diss1 Furosemide 4 screen 352 5 32287 5 1 71 No 11 0 diss1 Furosemide ts DiAevaluation IGA Furosemide bir B t dt Data Window Heb oslus al 2 Oycle A B C D E F G 1 K L Al JAN fa eilan HAN lt HA A e Fe APROG Tim 3 screen 2 Biacore 2000 5 91 5 3 ja Customized Sequence TEMP 25 7 DETECTION 2 1 F FL
5. 1 1 Langmuir with mass transfer Biacore92000 60 4 A TOS ER 4 3 2 Linked Reaction 1 1 VAE X XN LJUBSE a 1 1 Eea Run gt Run Application Wizard Kinetic Analysis 83 Kinetics Analysis Steps EE4 Kinetic Analysis Steps Concentration Series M Control Experiment be Mass Transfer nens Li
6. 0 22_um CMS 10x HBS fpi dO PBS HBS EP 10 mM HEPES 0 15 M NaqCI 3 mM EDTA 0 005 Surfactant P 20 pH7 2 241728 MAR HBS P 10 mM HEPES 0 15 M Nacl 0 005 96 Surfactant P 20 pH7 4 241133 MARIA HBS N 10 mM HEPES 0 15 M NaCI pH7 4 HBS N 10X 0 1M HEPES 1 5 M NaCI pH7 4 MilliQ97K C 10 45753 10 mM HEPES 0 15 M NaCI pH7 4 PBS 10X 0 1M phosphate buffer 27 mM KCI 1 37 M NaCl MilliQe2K C 10 15751 10mM phosphate buffer 2 7 mM KCI 137 mM NaCl 59 DMSO jEE G2 C pH7 4
7. lt GE erp IisgmEm T 159 0073 EET 3 25 DES TE e mail Tech PEqueom
8. aos 5 3 1 If Then SPP bound 20RU JLOB TERRIER BS 7072 ETERXS e wil blw Sequence 37 DETECTION 2 1 3 1 4 1 F FLnw 20 B BASELINE baseline 5 INJECT Antibody 120 s INJECT END EL 10 RESPONSE bound 5 QUICKINJECT 1 mM Gly HCI pH2 0 20 p QUICKINJECT END PL 60 RESPONSE regeneration 5 Hin 9 De y E m GENE For Report Point bound y in Flow Cel Subtra Compari ls Less Than Y Con stant y Value 20 Use Additional Condition THEN IF RelResp For Report Point bound In Flow Cell Subtracted Comparison Is less Than Value Constant 20 THEN Exit
9. 3 2 gt BBA gt TINY pH 5 NaCl aM 10 mM Gly HCI gt pH 1 5 HCI 100 mM Phosphoric acid 100 mM Formic acid 20 96 10mM Gly NaOH lt pH 12 NaOH 100 mM Ethanolamine 100 mM Ethanolamine HCI 1M EDTA lt 0 35 M Surfactant P 20 Tween 20 5 96 Triton X 100 lt 5 SDS lt 0 5 96 Octylglucoside 40 mM t VEU Acetonitrile 2096 DMSO 896 Ethyleneglycol in HBS buffer 5096 Ethanol lt 20 Formamide 4096 Guanidine HCI lt 5M Urea lt 8 Biacore 2000 4 39 18 Bulk Effect f 8 48 OO RB EX 9 8873 cle SXSSR Bulk Effect RU 2200 Bulk Effect o o a Biacore92000 40 4 AIE DIA ER 4 1 Y PIRE d S THE TEFHOECENREY ic PAI Run gt Run sensorgram
10. SEE Rack UG 0 5 pH pH Immobilization pH Scouting ERANI Do Immobilization pH Scouting EETL AAO pH pH _ 5200 ug ml 10 mM Immobilization pH Scouting
11. Temperature 5 1 Biacore92000 10 1 1 2 5 SPR Thermo_A 2 BIAmaintenance kit BlAnormalizing solution 0 5 ml 9mm R2F2 Tools gt Working Tools Normalize Dl Working Tools EJ Cancel Mormalizes the signal in all flow cells Use this procedure aj when the opto interface is changed b when the temperature is changed c when a new sensor chip is
12. 50 ml 4 ALEIRLIENS UZJZ ROBELTUVAL MBA C Bc seBytnlc 3589 U NS TED Md SIX E Dock Biacore92000 96 ESTATE 7 7 1 Tools gt Working Tools Prime Dock
13. Regeneration Add Analyte Biacore 2000 50 4 ALP 2e ES Ea Surface Preparation Regenerate i ES Regeneration Solution Add 10 mi Glicine pH 0 10 mid Glycine pH 2 5 10 mk Glycine pH 3 0 Remove 50 ml NaOH Injection Settings Injection Time ns mir Flow Rate 20 ulmin Position Rta Required volume 65 pl The door is unlocked until you press Start Help Start Cancel CM 0 5 1 20 lt 60u min pd Add Add New Regeneration Solution Mame nm HC Cancel Start Regeneration Scouting
14. 1 3 4 Blank NHS Ja Surface Preparation Flow Rate and Injection Time X min l min PO O 5 gj O O L2 jignd o 10 NM La 1 nqp Help lt Back Mest gt Close 3 Biacore92000 30 3 EDC Coupling Solution 115 NHS Coupling Solution Empty vial for mixing min capacity 200 pl 110 Ligand 75 Ethanolamine THERMO C y THERMO_A y 00 00000 o o 000000000 e000000 O O 9959929225528 o d 0 Make sure that the correct rack s are installed in the autosampler when you start the run Help Menu lt Back Next gt Close EDC NHS EDC NHS
15. 50 mM NaOH Biacore92000 22 3 Run Run Application Wizard 3 Run Application Wizard Surface Preparation EE EE t Start Analyte Recovery Binding Analysis e Templi Kinetic Analysis Customized Application Cancel The Surface preparation application wizard guides you through the essential steps in preparing the sensor surface for analysis including immobilization pH scouting ligand immobilization regeneration scouting and surface performance test Surface Preparation Start y Immobilization Immobilization in Surface Prep Regeneration Scouting Surface Performance Test The Immobilization pH scouting wizard guides pon through determination of a suitable pH For immobilizing ligand on Sensor Chip Ch Help Back Hest gt Cloze Immobilization pH Scouting Next gt Cea Surface Preparation Buffer Selection 10 mid Acetate Add 10 mh Acetate 10 mh Acetate Remove 10 mh Acetate 10 mM Acetate pH 4 00 Prepare ligand solutions in the buffers vau have chosen Help
16. 4 CC 200ul 1 BSA 10 mM 10 mM Borate 1 M NaCl pH 8 5 Biacore92000 3 20 5 200ug ml 10mM pH 0 52 pH pH 3 5 Immobilization pH Scouting 10mM pH
17. OK EH 2 3 ZALE HIE e 3X ABLE LI MAS FM RO Ctrl Break Biacore 2000 3 17 3 UHE 90 _ EN CM5 mU e 237 Fx IBIC ES S 7 x7 E N g CM NNS NE ULM PO a As YA T e E 69 Os EET NHS DST PRA ue UL KORB CEEI SRA 71 Vi AU CS S Abs CELS S735
18. Next gt y Standby Flow After Run Prime Before Run Start y sa BIACORE E x 32 Do you want to save current wizard template Save Save As Don t Save E Fale 7402 RIDE 2 UM RELCABS SI co AY DR KEND gl Don t Save 11 BO Save Ctrl Break 35 If Then Report Point MM a ci RR RelResp Mi RU AbsResp RU Slope RU s In Flow cell Jp pet
19. ugm Uy eb _ 30 1 Biacore92000 38 4 la n PITA bke seslt S HD SEIS tese SEDES DU 17
20. WOODS EEN 5s XU 1 1 Protein A A 2 ADO 3 2 re Vay by Cycle Vay by Cycle Solution Vary by Cycle Antibody1 OK y 2 IT Vary by Cycle Vary by Cycle Solution Antigen Vary by Cycle OK Moros i NS Vary by Cycle T Vay by Cycle Solution Vary by Cycle Antibody2 OK Biacore 2000 5 T4 Customized Sequence A INJECT Antibody 50 d INJECT END INJECT END INJECT Antigen 50 Le INJECT Antibody2 50 8 INJECT END QUICKINJECT 10mM NaOH 10 d QUICKINJECT END Help Menu jac Nest gt Close Next gt Ta Customized Cycles Run Order As Entered v Use Separate Vials for Differe
21. y RED 0 2 LE RE ir y View gt Biase Line 0 LE LC due Ma DOLAR AAN RMS lOs Biacore 2000 4 45 23 Command Queue Command Queue
22. Rack Base1 Rack Base a 1 3 R1A1 R1A2 R1A3 96 _ Biacore 2000 1 9 1 2 4 Command gt Set Temperature E Set Temperature X EG set Temperature OK Cancel Temperature 47 40 CO8BBH Cag4EL OK 7 4 40C
23. Regeneration Do x Ciria O EU ADO KH Biacore 2000 4 51 Regeneration Scouting Regeneration Scouting Finish sm Hen scau Results 1 3 10 mM Glycine pH 2D 1 3 10 mM Glycine pH 1 5 1 3 10 mM Glycine pH 1 5 Regeneration Analyte 80 20 pH Analyte 80 20 20 8096 2 90 Biacore 2000 52 4 IP SHERSS 4 2
24. Immobilization pH Scouting Time AbsHesp Slope Baselne RelRhesp Id ImmobPH 128748 Baseline Trypzin inhibitor ImmabPH 11153 5 2 10mHM Acetate pH 6 00 Trypsin inhibitor ImmabPH 128726 2 Baseline Trypsin inhibitor ImmobPH 111540 4 10 mM Acetate pH 5 50 Trypsin inhibitor ImmnhPH 128741 5 Baseline3 Trypsin inhibitor ImmnhPH 111538 3 10 mM Acetate pH 5 00 Trypsin inhibitor ImmabPH 128737 A Baseline4 Trypsin inhibitor ImmobPH 15125 0 3 10 mM Acetate pH 4 50 Trypsin inhibitor ImmobPH 13047 5 8 Baselineb Trypsin inhibitor ImmobPH 18542 7 2 10 mM Acetate pH 4 00 Trypsin inhibitor ImmabPH 13408 4 PreHegen Trypsin inhibitor ImmobPH 12835 8 571 6 Regeneration Trypsin inhibitor Biacore92000 26 3 Trypsin inhibitor g 1 mM Acetate pH 6 0 5 t 10 mM Acetate pH 5 5 8 M 10 mM Acetate pH 5 0 i 1000 10 mM Acetate pH 4 5 0 10 mM Acetate pH 4 0 1000 2000 3000 0 8 16 24 32 40 48 56 64 72 80 Time m View Ligand Es Overlay Plot de ee 1 5 0 C Table 15 O mM Acetate pH 4 0 Immobilization pH pH 9 9 Immobilization pH Scouting
25. S S a CERES O s Biacore92000 pl Rmax 100RU Rmax x xS Da RU
26. A 500 HEB z9 IFC amp 27 7 78S CUUDPS SJ zmosm 2UIUZERSREXUEXUKC ROIZ F a i S PA 3 If Then 5 3 2 35 Biacore92000 76 5 5 3 1 JDP 1 Fc2 2 Fc3 3 Fc4 No 1 No 96 96 el Run gt Run Application Wizard T Run Application Wizard E X Surface Preparation Analyte Recovery Binding Analysis Kinetic Analysis Start Use Template Lustomized A SEM OI MM MN t fr Cancel This wizard allows you to program and run customized applications For purposes that are not covered by the applicatian specific wizards Customized Application Start y He eg gm aes t dll
27. Enter Next gt 28 Next gt Ti BIACORE IN For reliable kinetic analysis pou should run at least 5 different concentrations per analyte Ignore 5 Ignore ma BIACORE AN For reliable kinetic analysis you should run replicates of at least one sample per analyte Ignore ID Ignore A x T Kinetics Regeneration Regeneration Flow Fate Flow Rate 30 pl min Regeneration Methad Dissociation in Buffer Single Injection Two Injgctions Solution Injection SG olution hn mk Glycine nH 1 5 Injection Time 30 3 Predip Needle Stabilization Time After Regeneration 2 min Help Back Next gt Cloze Regeneratio
28. Sensor chip Dock Dock Sensor chip 3 Dock I Undock Sensor chip Dock Biacore92000 LEVAS E4 1 Sensor Chip CM5 IDORRA a sd E A o lO a MA le SU DEEP Resear
29. Next gt Ta Surface Preparation Prepare Run E x THERMO 4A right hand rack position Position Min Vol ul Content B1 115 EDC Coupling Solution B2 115 NHS Coupling Solution B3 Empty vial for mixing minimum capacity 200 pl B4 110 Ligand B5 75 Ethanolamine Prime Before Run w Standby Flow After Run Help Menu lt Back Start Close Standby Flow After Run Prime Before Run Start Es BIACORE EE x 2 Do you want to save current wizard template Save Save As Dont Save EA il NA AO E a Aye Y Mind XT Don t Save 11 Biacore 2000 3 31 Save Results A ETr rr ES Save in Tanaka eca D test EA History Desktop My Documents File name immobilization My Network P Save as type Results blr hd Cancel Sqve y
30. 5 Cm TAREA pH amp LU SJ UJJ7 ARES RUSCclCcd U JS Too slow binding of ligand TONWEGDURO pH Biacore 2000 4 37 4 rude
31. Binding Using Capturing Molecule Biacore 2000 4 Next gt Use Flow Cell s Analyte Name MW Next gt Regeneration Method Dissociation in Buffer Single Injection Two Injections Solution Injection Time Next gt Fi Kinetics Mass Transfer Control x M Analyte Hame Bai rml Hu Antigen 5DDUD Use Flow Cell s The mass transfer control experiment analyses a single analpte concentration at a range of pre determined How rates for each analnte Contact time and dissociation time are Fi ed The shape of the sensorgram varies with Flow rate if the mass transfer processes are limiting Help Back Mest gt Cloze i Ti Kinetics Regeneration Regeneration Method Regeneration Flow Rate Flow Rate ln u mir C Dissociation in Buffer Single Injection C Two Injgctions Solution Injection Solution mM NanH Injection Time 0 s Stabilization Time After Regeneration 2 mir Help lt Back Mext gt
32. Surface Preparation kK Run gt Run Application Wizard Surface Preparation Start T Surface Preparation Surface Preparation Steps E X Surface Preparation Steps Immobilization pH Scouting Immobilization Immobilization in Surface Prep Fiegeneration Scouting ss i Surface Performance Test E Helps nu to check the binding capacity of the surface and that the immobilized ligand withstands regeneration conditions Help Back Hest Close Surface Performance Test Next gt y Ta Surface Preparation Analyte Settings E ajx Analyte Settings Mame Antigen Injection Time 2 min Elow Hate 2d pl min Run Settings Number of Test Cycles 5 Use Flowcell 3 Help Back Mast gt
33. 0 Mind Dont Save y Biacore 2000 4 Save in E Tanaka lt 53 ck E3 D test nts ter File name Mass Transfer la Save as type Results bir Y Cancel Sdve My Network P n Ctrl Break S mass tarnsfer_test2 Results Anale B54 BSA 100u3 ml 140 120 100 5 ul min 15 ul min Response T5 pimin 30 B g0 120 150 180 210 240 270 30 Time amp Baseline Level Wass Transfer Linked Reactions 59 Go
34. DELIS IU EE oe 72 J9 7 52 100ug ml 10mM Borate 1 M NaCl pH 8 5 NHS pH 8 5 DMSO Instrument handbook Biacore 2000 3 al 3 1 1 UY RAMORO pH CM5 CM CM
35. Id Antibody1 1 Antibody1 2 Biacore92000 80 5 Run Order Random As Entered Step Next gt a Dus a ami Lm Fuck LL EE 160 Antibody 160 Antbody 160 Anirksdr Antibody Auritibody 1 Antibody TH Antbbsdy 16 Antbody Antibes Antibody MICRO THERMO A eeeeeees 20202001 00000 00000 Oooo OO ooooQt 4080006000 oocom OO0000O O BD Ooo 2600009000 202500070 Any ergo you make atha rack stg rel eoru e Hep Mem fr Beh iones ee Next gt y Standby Flow After Run Prime Before Run
36. Response 2 Response 1 NHS Reponse 2 Response 1 Biacore 2000 3 23 3 1 3 Immobilization 3 1 2 Zea Surface Preparation Immobilization Procedure Immobilization Procedure t Specify Flow Rate and Injection Time Help lt Back Hest gt Cloze Aim for Immobilized Level Next gt Tj Surface Preparation Aim for Immobilized Level Flow Cell in Ligand Hame Wash Solution a L3 T ETT iamgowwusom EN o Help Back Hert gt Close Ligand Name Target RU Wash Solution L 23 2 RS e SAR FA N ds JU fe 388 50mM NaOH Blank 13 Next gt
37. Dock HBS EP PEDIR C Prime y 2 Thermo_A R2F1 BIAtest solution 1ml 9 mm R2E1 E2 EX E4 9mm V Tools gt Testtools gt System check Biacore 2000 Ch Test Tools quality ch System check Checks detection unit How cell factors pump function autosampler performance and IFC sample loop and valves eck System check evaluation Total run time le about 32 minutes NOTE This procedure should be run at 25 C with a new Sensor Chip CM5 Last Aun Time 0 Feb 06 18 33 Start MES fy 40 Response 9 VIR PRU NE ERA 99 System Check To evaluate the System check click the E Evaluate valuate button Biacore92000 100 LATA BIACORE 2000 System Check Name Created By BIACORE 2000 Control Software Version 3 0 7 Beta Instrument id 2076 Configuration IFC3 with Recovery Run Date 22 Mar 1999 Evaluation
38. E His Tag Fusion Protein Ni D Sensor Chip HPA EROIICADITYJUE C18 monolayer es Sensor Chip L1 FRAMES E d55 T UsBEIUL Ci SZVwWTLXCITMABLDULVA AOTW J UVfsevoyelgtS cC UV lcUCmNDIUISSCCC 88 bilayer C Biacore 2000 1 5 1 2 2 TV Y JEENE Dock Working Tools Prime Ba Working Tools Start Start Prime Exit Biacore92000 6 LEVAS A 5 Prime D Prime Tools Working Tools
39. Biacore 2000 1 7 1 2 3 Command Rack Base l Rack Base i l Rack Base renmo ce THER A None MICRO THERMO A THERMO THERMO E E M 000 000 OK UU VU Tg Ss e 2 V2 ERACI P MIc oU Cli fixe 6 Biacore92000 8 1 6 Rack Base1 Rack Base2 INSECTA mers A E TIT TIA A mm 9 mm 1 5 ml 2 ml 16 mm m A E Y Thermo_A Thermo_A Thermo C Thermo C Thermo A A D Jl EA F1 F2 F3 F7 Thermo B
40. 46 ross 9 AEE e e e eeo 16 IMM rss 33 c e e mnne eeGee6eveece 2 eS lcccccc 37 38 META A m m m m t m n n ng n n 19 20 21 10 e o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o 98 3 5 e o o o o o o o o o o o o o o o o o o o o o o o o o o o e o o 0 0 94 e e e t om om om om om e t om on o9 om o9 om ot t n n 9 o9 m m t ng n9 n9 n9 n9 9 94 41 T e o o o o o o o o gt o o o o o o o o o o o o o oo o o o o o o o o o 3 PVH FYVITDEXOBL f s e 9 9 om om om om n nm nm nm ng 95 Biacore92000 104 DS y ng 95 I DID E ULM DN DE EO 57 228 AEE E ac pu 1 INS AR EAS E A A OE vds A E AECA SRA AA E 17 84 4 amp 3 3 788 0 Bl EF SU UR eC CIN URS 2E ER DECORE NEU uh DS e ATAN a AE e WC RT DN 20 REED E 101 AUD Eb 4E O AA T A O O E A va VET deo A dn ce 46
41. Standby Flow After Run Prime Before Run Start Fi BIACORE EE xi 2 Do pou want to save current wizard template Save Save As Don t Save 1 iEn cp AO mp Dont Save 11 Biacore 2000 4 49 saveResultsas 3 xl Save in Tanaka 9 tct E3 3 m test My Computer iu File name regeneration scouting Save as type Results blr y Cancel Save My Network P Ctrl Break y Regeneration Scouting Response 3 4BR9m00 T0255 Begeneration DSA 100 20 31 0 100 Um Add A4nalyte Finish Analyte bound in last injection 31 0 AU Total amount analyte bound 31 0 HUI set ta 100 Help Status dle
42. ia 3 Le Edit gt Add Report Point T Add Report Point Sensorgram Fo 1 DE dd Time es A 8 Cancel window E ld baseline 4 Baseline A TE e Apply to all curves Id DIAS FAQS Lo 0 RU Baseline RelResp 0487 0 0 Biacore 2000 2 15 Injecilan of several concentration ol Sucrose Fr Tma Abe Sipe Bmore AsRosp ld 655 1249559 0 06 Yes baseline RelResp RU Injection of several concentration of Sucrose 40000 35000 30000 25000 20000 Response 15000 10000 5000 0 100 200 300 400 500 600 700 800 Time Fc Time AbsResp Slope Baseline HelHesp Id 12495 9 0 06 Yes 0 baseline 1 127 5 154229 0 02 No 2927 0 2 sucrose 1 298 5 18309 7 0 08 No 5813 7 4 sucrose 1 4725 24
43. DEP OK Delete Command Ha w2 blw Sequence n Edit Command CtrI E Report Point Gtri R Inject Ctrl I Comment Ctrl B lo BASELINE baseline 5 Wait GtrtW Transfer otrhT A INJECT Antibody 120 y Ster D F FLOW 20 Mix Ctrl M Flow Path INJECT END Flow Ctri F B 10 RESPONSE bound 5 wash IF bound Subtracted RelRe H Then Y EXIT CYCLE Cycle Settings Ctrl Y END IF Keyword Ctrl K QUICKINJECT END Save Wizard Template s B T RESPONSE regenerati Print Sequence Print Wizard Template Q IF regeneration Sample Rel T Delete Command 25 QUICKINJECT ma P3 QUICKINJECT 10mM Save Wizard Template Otrl S D Ctrl P QUICKINJECT END Biacore92000 82 5 33 1 Solution
44. 270 A 100 150 200 250 300 350 400 450 Time s Apparent constants according to fitting model 1 1 Langmuir binding ka 1 2e5 1 Ms kd 4e 4 1 s RMax 338 HU KA 2 9e8 1 M KD 3 4e 9 M Concentration Series Baseline Level ka 1 Ms ka FEBR SS AER 1 5 Ka 1 M Kp ERREEN M Rmax RU 1 1 binding pum Baseline Level N KN 7 C 9 O rabbit lgG Results Flow Cell E Baseline Stability 2 o Biacore 2000 5 69 5 2 RISEBGCE aio Run gt Application Wizard E Run Application Wizard E x Surface Preparation Analyte Recover E Lse Temolate Kinetic Analysis Ose lemplale Customized amp pplieatian Cancel The binding analysis wizard guides pou through the steps involved in testing binding of analptes to the immobilized ligand Binding Analysis Start es Binding Analysis Assay Principle Q em Binding Using Capturing Molecule Eaptunng Parametere
45. Ta Detection Er Detection made Reference subtraction Erre m x e none c E Fe1 2 34 OK 19 19 Detection 1 3 E Detection mode Men i subtraction C El Fc 1 Cancel none c E rez c El Freies pa c e Fes C Z Fc 4 9 1 U2 0 675 2 1 Fc1 2 3 4 De Detection mode E Reference c E F
46. 1 1 1 1 binding Biacore92000 64 5 5 Customized Application 5 3 5 4 Kinetics Analysis 5 1 amp Binding Analysis 5 2 jy OL Customized Application 5 3 Customized Application 5 4 5 1 Run Run Application Wizard 7 J v 7 3 5 SH Run Application Wizard E X Surface Preparation Analyte Recovery ding Anal se Template Cancel The kinetic analysis wizard guides vou through the steps In de
47. Ctrl CET Break i RU Amine Coupling Ligand 50000 45000 40000 35000 i 30000 25000 R 20000 15000 10000 0 200 400 600 300 1000 1200 1400 1600 1800 2000 Time s Fc APROG Time AbsHesp Slope Baseline RelFiesp ld Blank InjectTime Ligand ame FlowAndTime 272 5 160727 0 04 Yez DO Baseline Hn 7T Ligand 2 Flow ndTime 825 5 316204 8 0 09 Mo 132 0 EDC HHS Ho 7 Ligand 2 Flow ndTime 1368 5 20625 6 0 36 Mo 4552 8 Ligand Ho 7 Ligand Flow amp ndTime 1918 5 205413 0 03 Ho 4468 6 Ethanolamine Ho 7 Ligand c proteinA_immob050616 Results x sensor Chip CM5 Ligand Response 1 Response 2 Pracedure bound final 4465 6 Injection time 7 0 min flow 10 pimin Immobilization Biacore92000 52 3 HE 14 2 Response 1 RU Response 2 RU NHS Amine Coupling Ligand 15000 10000 0 200 400 600 800 1000 Time VIV ROREEUL TUSASVRIAD F Y ROM JEE TI Y UP Y
48. RU Is Greater Than Is Greater Than or Equals Is Less Than Is Less Than or Equals Equals THEN Exit Cycle Do Exit Method Command Block ff Then Biacore 2000 5 89 5 4 Biacore vw A B B A RU A EST MER RU vs C BE 7o 0 7 Qm AA A A
49. 5 79 NN A ON ADO S 10 After ld OKk Y Ta Customized Seguente EA DETECTION 24 3 F FLOW 20 B de BASELIME baseline 5 A INJECT Antibody 120 Fs INJECT END El 10 RESPONSE bound 5 9 QUICKINJECT 0mM Gly HCI pH2 0 20 QUICKINJECT END PL 1 amp 0 RESPONSE regeneration 5 Help Menu s Back Next gt Ta Customized Cycles Hun Order As Entered lv Use Separate Vials for Different Solution Variables Antibody l 1 Antibody2 1 Antibody3 1 Antibody4 1 Antibody5 1 Antibodyb 1 Antibody 1 Antibody8 1 Antibody9 1 Antibody10 1 Antibody11 1 Antibody12 1 Antibody13 1 Antibody14 1 Antibody15 1 Antibody16 1 Antibody17 1 Antibody18 1 Antibody19 i D Menu bd lt Back Nest Close Repl Antibody
50. Biacore 40 TEA E 1 AY 20 kb 4 AY 76 kb A SA UT 1 20 kb 1 5 90 kb 5 5 150 kb Biacore 2000 103 RI 45 40 4 17 19 21 27 33 puyjwJUuJg wvyhk 9 O o o o o o o 25 5555555555 19 o eee 55 s st st n 9 t n 9 n9 n9 1 17 84 84 97
51. aks 9 0 Command Queue al ES Ele Edt Run 9 2 QUICKINJECT R2E1 10 4 QUICKINJECT R2E1 10 4 QUICKINJECT R2E1 10 C FLOW 10 Cj MIX R2A1 110 EF FLOWPATH 1 2 3 4 4 QUICKINJECT R2E1 10 Edit gt Delete Edit Insert AE 24 r EL X di Command Flow E Set Flow Flow BE ul min Refil Cancel Ae 25 RE H9 D UE ES Command gt Flow Path 5 Flow Path Elow Path E Detection mode 1 2 3 4 Essi a Is Fc E Fc 1 2 3 r E C Fc 1 2 3 4 c amp res OK Biacore92000 46 4 AIE 7 84H 26 ANDO cpi ADO c XR icc dels ll Command 7 Stop Inject Injection Stopped A Remaining 5 ample E Dispense remaining sample to njection x njection in
52. tie 15 O Ata RU 16 DEA NHS 5ul min 7 4292 00739 71 3 5ul min EL 5ul min 7 Biacore92000 34 3 Surface Preparation Rack Positions 115 EDC Coupling Solution 115 NHS Coupling Solution Empty vial for mixing min capacity 200 pl R2 B4 210 Ligand R2 B5 75 Ethanolamine 50 50 mM NaOH o ooeoo 0 OOOO 5o5 5 d eecc i 9 ee ee t o 0000 OOOO 00 000 00000 QOQDO ET Make sure that the correct rack s are installed in the autosampler when vou start the run Help Menu lt Back Next gt Close Next gt i Ti Surface Preparation Prepare Run THERMO_A right hand rack position Position Vol
53. ON 0 1R6 AIM AR a ME REDA ERAI KEI MiliQe2K ERE 6 30 Prime 2 3 Flush IFC 3 Rinse IFC 20 UOJV amp 27 V DBA CiU 8 Desorb 1 IFC BIA maintenace kit BIAdesorb solution 1 0 5 9 SDS BIAdesorb solution 2 50 mM Gly NaOH pH9 5 22 NE Thermo_A DYZ TREN MilliO R2F3 BIAdesorb solution 1 3ml R2F4 BIAdesorb solution 2 3 ml 37 Desorb BIAdesorb solution 1 4 D MA s 20 Desorb MiliO K Prim
54. Start y Ti BIACORE 2 Do pou want to save current wizard template Save Save As Don t Save A lO A A A OS lA NT Mind Don t Save 11 BS y Save y Ctrl CER Break Biacore 2000 5 81 31 2 OK ECTTTTERCCONNNNEENEAAAM a T INJECT END E FLOWPATH 4 21 KINJECT Sameie LE KINJECT END 32 l
55. A E A A AN LR Os d 1 eS Ce 52 64 89 CON E Dens X ou 15 E A A A E 5 6 94 96 97 107 a NES e 9 0 o 9 9 9 9 9 oo o o o o 9 9 9 9 9 9 ee ee 96 s 9 9 m om om om om m tm m t nm n 20 TORTA D nai E IK UMS DES e 4 2 O E A A 10 4E TUS 907 87 C9 96 78 og F SX e RW o INC INDIAD LED Oe A DM HEC DR ORT A A E ARA aX GNE OS SRL Te ees 19 21 MUR ry EX ru MM DEEP 17 84 3 4 AE AA AE EAS Wesce Xe Do CIO A A A RATA A sug 17 JI PLULIZRIS Vr 2 0200 I1 0 oeeo 14 15 44 E J JA e e e e m m om om om om om om o9 o9 o9 o9 o9 o9 o9 o9 o9 o9 o9 o9 o9 o9 os n n n t t9 t9 t 5 5 5 1 96 Biacore 2000 105 t t mm m tm tm tm t n n n 14 A e 40 uL A A a a e ad a a a a a a a a a a e e aa 98 Biacore92000 106 INDEX A Aim for immobilization level e o o o o o o o o o o o o o o o o o o o o o o o o oo 28 B BIGINJEC ooe o oo ooo ooo ooo ooo ooo oo o ooo
56. INJECT OGO 21 BE xl J Injection Injection in Fe 1 2 T Start Injection P KINJECT Cancel Cancel Sample Position R24 Volume 40 ul Contact Time 120 Consumption More gt gt i Dissociation Time Start Injection Biacore92000 42 4 AI ENDRI Response y BERHO PADRE X NUUS S o PA Al Command gt Inject HESHEEADL Star Injection Response 22 21 INJECT 5 325Ll 30Ll CMT BEHEEADSS 0 fase ADS
57. MilliQ Tools Working Tools Prime Close 16 mm MiliQ 3 ml R2F3 Tools Working Tools Close Biacore 2000 6 95 6 2 Command gt Undock Undock Sensorchip i Biacore i AN A ESA 6 3 50 ml CEN 4 C FE DNA Do
58. My Computer Windows My computer vy LOPE O US Bia C Bia Users ex Bia C ojx File Edit iew Go Favorites Help E tuuc d gx Back LR Bia C Blaewal bia lib Compad Demo hd abject s selected E My Computer BIA Users 1 lx File Edit Wiew Go Favorites Help a shortcut recte Tut Delete 4 Wave Sound Rename Text Document Properties WordPad Document 29 Bitmap Image My Computer 3 Briefcase Bia C v A Users Work Offline Close Demo Hosoya lwata san File New Folder Enter Biacore 2000 102 8 TIBIO Explore Start gt Program gt Windows Explores C Bia Users File gt New gt Folder Enter 39 Bia Users
59. 50 000 Da 1 000 RU 1 J 7 20 Boy 20 000 Da JERAR Rmax 20 000 x 1 000 50 000 x 1 400 RU 10 000RU RU vs C kaka REES Ko YA RIVAMRUZTIV 3 Bliss RU 8888 RU 200x1 Sx RU 1000 x 1 S x S 50 kDa 100 kDa 1 200 x 1 1 x 50 000 100 000 100RU 1000 x 1 1 x 50 000 100 000
60. HEL e MH A ii NEED ua A i O a E ni FZV TREN Biacore 2000 2 1 1 1 2 BIA programs BIACORE 2000 Control Software REJBIACORE 2000 Control Software Ww Blh COME JIC Cunirel Elhmu wn OOk Epiwupe Ernaur a tl Edo Jis zomend Em Too goys Jep gr wa EE E A eel Elbe erem e a El T Primaryzmab 1 secnndzMzB 1 FM E cx m SS nem bar z AA r i r d E mm a H HU B i I ZAC H 4 lm da l E OP gt 11 rd I 5 E y 3 Sensorpram minina lm i k Im IP EE p n u al 131 11 1 F I 21 111 Lime F Tins darmkau PO Toa bm alere 0v ia md apipi 4125 lanh A nli Tua Dn AFI HF Put iili 3 3 4d DOOplupi mr H 144111 11 HI THa Il Hail HF HAt 1 ll 1 5343 IM 1 php d TE TN Yrs II HANILA hah 1 dash 12014 4 snap dT ddubio DJ Yes D nsFLHE Mab wabi 141 Hepnripunt tahle pinup 115 5 AL E 0 10 Tuu HD pinir Mu Wal I 1 4 FIL id Fr rH dui Tins in META HF TILI ri al paa ri A E n Ua Ext A A A S Fhrew limi Fir ull ET A TER x 05 c I m raid A A LAS A ELLA Event log Tahle E ce es I
61. Number of Sample Injections 4 o 3 Biacore 2000 5 71 C Binding Cycles Sample Repl Run Order Random As Entered Step Next gt Regeneration Method lt Back Next gt Close 5S1 1 S1 2 Ta Binding Analysis Regeneration Regeneration Method Regeneration Flow Rate C Dissociation in Buffer Single Injection Flow Rate 30 pl min C Two Injections Solution Injection
62. l T Aldehyde Coupling Wizard Template E E Wizard Type Customized Application Edit IL Notebook Aldehyde Coupling be Bun method Far immobilization af ligands containing aldehyde groups to hydrazide modified supports The method can be used for e g proteins with carbohydrates where Save s aldehydes are formed hu oxidation before coupling Print See Bl amp applications Handbook for more information Help Close Edit Customized Application 5 3 Gave Biacore 2000 5 85 5 3 2 5 3 1 lf Then
63. c o amp 2 20000 15000 10000 5000 0 100 200 300 400 500 600 700 800 Time s Fc Time AbsResp Slope Baseline RelResp ld 1 1249593 0 baseline 1 127 5 154229 0 02 No 2927 0 2X sucrose 1 298 5 183097 0 08 No 5813 7 4 sucrose 1 4725 240101 0 24 No 11514 1 8 sucrose 1 6485 353049 0 92 No 22808 9 16 sucrose i i La tU ld Run Stop Sensorgram Biacore 2000 14 2 2 1 2 RU i View Reference Line ARM PO 0 U
64. Tools Working Tools Prime Flush pe Rinse 2 Tools Service Tools Unclogging IFC RA 20 STR EN J 7 4 2 Biacore92000 98 ND SNP 006 2 7 3 Z CN Rabbit IgG 1 25hg ml Tools Service Tools Unclogging 7 4 CM5 HBS EP 9mm BIAtest soluton BIA maintenance kit Desorb Sanitize
65. 4 5 2 1 Sucrose 296 Sucrose 496 Sucrose 896 Sucrose 1696 Sucrose 2 1 1 100ul Ju E ip Run gt Run sensorgram BN Detection mode Er Reference OK subtraction CE Fe c S Fe12 po S c E rez c El ret24 foe 7 c El Fes c El res Detection mode OK Biacore 2000 12 2 1 100ul min 0OK y y Iib sso command gt Inject E9U y 933 Injection MEC E O 10ul Start Injection y Injection of several concentration of Sucrose o o LY tc Biacore 2000 2 15 RU Injection of several concentration of Sucrose 40000 35000 30000 25000 o
66. _ Heb Menu Bo Mest gt Cloze FLOW 20 y ri VESINE CADE ES Ta Cycle Settings dc SetTemperature DK E Set Detection Mode Cancel Detection de Setting the detection mode will algo set the initial Flow path Detection Fc1 2 3 4 Ref 2 1 3 1 4 1 OK Biacore 2000 5 7 Ti Customized Seguente FE FLOW 20 FLOW 20 Eri Inject Injection Mode Normal injection INJECT M Sample Cancel Solution Antibody Vary by Cycle Volume 12d ul Yarn by Cycle Help Options Predip Needle mos Injection Mode INJECT Solution Vary by Cycle Volume OK INJECT Antibody 120 E INJECT END i ha Inject PA Injection Mode MAA ESO TRI EN Sample Solution E mM Gl
67. 30000 28000 26000 24000 22000 20000 18000 16000 14000 0 immob_ anti alb Results 3 Aim 49 5 177221 3 Aim4 71 5 16878 7 3 Aim 77 5 174274 96 14 3 Aim 91 5 188049 100 47 3 Aim 1215 219937 113 46 3 Aim 26513 3 3 Aim4 17706 2 Response 1 2 14 Biacore92000 36 3 16 Aim for immobilized Level NH5 Sensor Chip CM5 Ligand Response 1 a En o s bound fina mmme f Too slow binding of ligand The surface in flowcells marked with has not been activated an Immobilization Too fast binding of ligand
68. Cloze SAR 57 1 2 2 Biacore92000 58 4 A TOS ER T5 Kinetics Rack Positions ul Antigen r1 E 10 EH Ns OH THERMO C THERMO A 60006000090 200000 oh b hb hb hb hb E LEN ON M EE E EE O O O Make sure that the corect rack s are installed in the autosampler when vou start the run Hep Menu lt Back awt gt Close Next gt y T Kinetics Prepare Run THERMO A right hand rack position Position Min Vol pl Content A1 320 50 pol 1 phi Antigen F3 275 10 mh NaOH Prime Before Run V Standby Flow After Fun Help Menu Back Start Cloze Standby Flow After Run Prime Before Run Start y CES BIACORE 2 Do you want to save current wizard template Save Save As Dont Save i
69. Mame Ligand Elow Rate 5 ull min Injection Time E min Stabilization Time fi o min Help 4 Back Hest gt Cloze Direct Binding Binding Using Capturing Molecule Direct Binding Next gt y Biacore92000 70 5 Ti Binding Analysis Injections Settings Use Flow Cells ZAT MEC Flow Rate 20 pl rnin Number of Sample Injections y m 4A Sample Injection Time 2 min Wait After Injection 2 5 min Help lt Back Next gt Close Use Flow Cell s Flow Rate 10 20ul min Number of Sample Injections 29 Injection Time 0 5 3min Wait After Injection FE SEE GS Fes 2 5min Next gt 29
70. Solution 10 mM Glycine pH 1 5 Injection Time 30 s Predip Needle Stabilization Time After Regeneration 2 min Help lt Back Nest Close Dissociation in Buffer Single Injection Two Injections 1 2 2 Next gt Biacore 2000 12 5 Ta Binding Analysis Rack Positions THERMO_C y THERMO A ODO e amp o o0 eooo 0 ee ee 9 eM TE e 5000 60 0000 NE NE EX Make sure that the correct rack s are installed in the autosampler when you start the run Help Menu lt Back Next gt Close Next gt Ti Binding Prepare Run lx ITHERMO A right hand rack position n Position Volume ul Content A1 60 Sample 1 A2 60 Sample 52 A3 60 Sample 53 A4 60 Sample 54 A5 60 Sample S5 A6 60 Sample S6 A7 60 Sample 57 AB 60 Sample S8 A9 60 Sample S9 A10 60 Sample 510 B1 60 Sample 511 B2 60 Sample 512 B3 60 Sample 513 B4 60 Sample 514 B5 60 Sample 515 B6
71. lop A Slop vs C B Calibration Curve A A A A A A B B A S ASUSA mS 1 20 30 40 50 60 70 so an 100 410 Concentration unite mil Biacore92000 90 5 Customized Application 5 3
72. 500 RU Sara E 100 500RU Biacore 2000 3 19 3 1 N g CM NHS N NHS CONH COO COD CONHCH CH OH NHS BR 1000 50 dq usc cos EDC N ethyl N 3 dimethylaminopropylicarbodiimide hydrochloride NHS N hydroxysuccinimide 1 M ethanolamine hydrochloride pH 8 5 EDC NHS 10 ml MilliQe 200ul 7 mm 20 1
73. Break Biacore 2000 4 55 33 BIACORE 3000 Control Software on COMI regeneration test Sensorgram hm File Edit view Command Run Tools Window Help cg 3 A ZE Hn k amp Cycle 1 Curve Sensorgram Fc 3 Surface Performance Test Baseline Level RU 18750 18734 18718 18702 a 18686 18670 0 1 2 3 4 5 6 3 Performance 64 5 187061 0 3 Performance 1445 18859 2 0 Analyte BSA 200ug ml 1 min Regeneration 25mM NaOH B0 s Surface Performance Test Sensorgram Fc 3 Time 3635s Response 18715 3 RU 0 0 Timestamp 15 Oct 38 17 52 57 0 0 Set Temperature 25 0 C 0 0 Temperature 25 00 C 0 0 Rack 1 Left Thermo C 0 0 Rack 2 Right Thermo amp 0 0 Rack A Reagent none Baseline Level Response Level Flow 0 pl min Flow Path Temperature 25 00 Detection Mode Single oU 2 v amp lcla 2e 0 c Uic D BOB MA BIE IC ERIS JA GER C do Baseline Level Response
74. QUICKINJECT 5 325Ul 10u Ea o COINJECT 2 Sample 1 0 100ul Sample 2 0 100ul BIGINJECT Er E 30pl INJECT Biacore 2000 4 43 Run gt Start New Cycle Edit View Command Run Tools Window Help Service3 1 Start Mew Cycle o LA La Stop Sensorgram Run Application Wizard Run Method Data Collection Rate Command Run Tools Window Help Seryice3 Er File Edi view File Edit Run 2 S Ej Run Stop Sensorgram File Save As Save in Tanaka e 3 tex E3 History t Desktop My Documents My Computer TAS E i File name tesi Y My Network P 5e Save as type Results br v Cancel M Sav
75. pH pH NHS pH4 pH4 5 NHS pH8 5 pH4 pH pH pH4 5 DSC 100 Y View gt Wizard Results 11 View Wizard Template Ta proteinA_immob050616 Wizard Template Wizard Type Surface Preparation Notebook Save s Print Help Edit 7 1 wg Sc COROT a E C
76. Back Next gt Close pH Next gt Add Remove y Biacore 2000 3 23 3 Surface Preparation Ligand Settings Name Injection Time 1min Flow Rate 10ul min Use Flowcell Next gt ui ourface Preparation Wash 2 Wash Method Two Injections Solution 50mM NaOH Injection Time AUS 30s Next gt Biacore92000 24 3 Position Vol f Content 50 Ligand in 1mM Acetate pH 6 00 50 Ligand in 10 mM Acetate pH 5 50 50 Ligand in 10 mM Acetate pH 5 00 50 Ligand in 10 mM Acetate pH 4 50 50 Ligand i
77. Cloze Name Jo pe Injection Time Flow Rate Number of Test Cycles 5 Use Flowcell Next gt Biacore 2000 4 53 Ja surface Preparation Regeneration Regeneration Flow Rate Flow Rate 20 u mir Regeneration Methad Dissociation in Buffer Single Injection C Two Injections Solution Injection Solution 50m NaOH Injection Time 30 3 Predip Needle Stabilization Time after Regeneration 2 min Help Back Hest gt Close Regeneration Method Dissociation in Buffer Single Injection 1 Two Injections 2 2 Solution Injection Time Next gt surface a Rack Positions E x Position mavan a R2 Al E Antigen R2 F3 300 S mhM NaOH THERMD E THERMO OOOO eeosoct 0 OOOOles s52590 eeceoi 9 90000 883280 0000 0000 TERS E LAE NE E tt Make sure that the corect rack s are installed in the autosampler when you start the run _ H
78. acce 85 89 6 TL RT A A A E 94 O EHE 94 6 2 Il 95 6 3 kk 95 SS lt 96 Y ES Ar EN Dc PN 96 7 2 dp S EA O Je tte t et t Gre te kk 97 7 3 lk 98 VO I ATAT T i Ni A ii 98 cc 101 E N A ESANA TE E TEE 103 Biacore 2000 1 1 1 1 1 1 1 1 gt gt gt gt MA gt Windows biacore Y 30 Y Biacore 2
79. p min Dissociation in Buffer t Single Injection C Two Injections Solution Injection Solution 10m NaDH Injection Time 30 3 v Stabilization Time After Regeneration min Back Henst gt Close Regeneration Method Dissociation in Buffer Single Injection 1 Two Injections 2 2 Solution Injection Time Next gt Biacore92000 Hl at 62 4 HENX Ta Kinetics Rack Positions x Position R2 Al 370 50 ug ml 1 275 10 mM NaO Ai Antigen uM y THERMO_C y THERMO_A y o o OOOO o OOOO OOOO Make sure that the correct rack s are installed in the autosampler when you start the run Help Menu lt Back Next gt Close eoo os 0000000000 000000000 ODODODOJDO 10101010 060000000000 O O O Next gt Ti Kinetics Prepare Run ES xj IHERMO A right hand rack position Position Min Vol pl Content A1 370 50 ug ml 1 piM Antigen F3 275 10 mM NaOH Prime Before Run v Standby Flow After Run Help Me
80. 60 Sample S16 B7 60 Sample 517 B8 60 Sample 518 Prime Before Run Iv Standby Flow After Run Help Menu lt Back Start Close Standby Flow After Run Prime Before Run Start y 2 Do you want to save current wizard template Save Save As Don t Save Enlr TAY PAIUDTDA ES IDA RE UCA a NE a IDR Dont Save 11 So l Biacore 2000 5 73 Squve Ctrl CET Break y RU S1 Rep 1 15200 15000 14800 14600 14400 14200 14000 13800 13600 13400 Response 200 Time 3 Analyse 51 5 14917 9 0 02 Yes 0 Baseline 51 3 Analyse 111 5 150153 0 24 No 97 4 Sample Hesp1 S1 3 Analyse 2625 1497239 0 05 No 55 0 Sample Resp2 S1 3 Analyse 401 5 148898 0 12 No
81. docked and maximum sensitivity is required Total run time is about 5 minutes Last Aun Time 12 May 99 15 30 Start 8 Normalize SPR O Biacore 2000 2 11 2 2 Do 3
82. 010 1 0 24 No 11514 1 8 sucrose 1 648 5 353049 0 92 No 22808 9 16 sucrose 2 1 3 File Save File Save As 313 Save in Tanaka e E History E Desktop My Documents My Computer TAO E File name tesi y Save as type Results blr E Cancel Ze C NBia Users Save DO My Network Ro Biacore92000 16 2 2 2 739 DEN File gt Print Ti Print Results E x Printer hip deskjet 990c series O Cancel System Information ly wizard Results v Table ss dd Event Lag A Setup Al Cycles Draft More gt gt Graph RP a ar ier PAN Table Event Log All Cycles Fit to Page 1 Draft Wizard Results
83. 28 1 Baseline Shift 51 Resp1 Resp2 Biacore92000 74 5 5 3 Customized Application 1 If Then 5 3 2 D Sequence 30 Ta Customized Sequence ta Customized Sequence 1 DETECTION 2 1 3 1 4 1 F FLnw 20 EB dy BASELINE baseline 5 A INJECT mouselgG 40 INJECT END B 10 RESPONSE bound 5 el QUICKINJECT 1 mM Gly HCI 20 QUICKINJECT END PL 10 RESPONSE re
84. 390 0 625 pon 4 17 nM Rabbit IgG F3 400 10 mM Glycine pH 1 5 Prime Before Run v Standby Flow After Run Help Menu lt Back Start Close Standby Flow After Run Prime Before Run Start Ed BIACORE 2 Do pou want to save current wizard template Save Save As Dont Save IER LOECHSI AB ZIOOSAeT7Z7Lu imclucCbhizsoDco2o0 20wWT D Dont Save 11 Y m Biacore92000 68 5 Save Results As 1 E 324 xl Save in a Tanaka 8 er En D test File name Kinetics Save as type Results blr v Cancel J da Sqave Ctrl CER Break 1 1 binding Ti rabbit leEG Results Analyte Rabbit IgG hal Rabbit IgG My Network Em
85. 42 Binding Analysis UE a e es ANE AMETE CD a e a a a a SL a dal 8 CR RE T 64 69 C Close a e aroda is UE a da WM e en e UA TEC ae anas ds ana UE O a la an an EA ORT UNS M ES 94 CM5 oeoo ooo ooo ooo 5c 2 5 e 5e 4 17 21 COINJECT oboo oo ooo ooo o ooo ooo ooo 42 Command QUeUe rr ross 45 Customized Application e o o o 110770202 o o o 64 4 6 89 D Desorb WU aS e dE A A o E a o a De CET a A A qe 96 Dock e o o o o o gt o o o o o gt o gt o o o gt o o o o o o o o o o o o o o o o o o o o o 3 E Extra Cleanup e o o o o o o o o o o o e o o o o o o o o o o o o o o o o o o o 46 F SE Y aa RA O A rop ib ve dea AA A 96 H GPA aoe Saa OS SM DS MO A AAA 4 84 1 Then 0 0 2 02 02 02 oo ooo oo oo oeoo ooo 85 88 Immobilization e o o o o o o o o o o o o o o o o o o o o oo oo oo o o o o o 17 Immobilization pH scouting e o o o o o o o o o o o o o o o o o o o o o o o o o o 21 INJECT eooo o o o o o oo o oo oo ooo ooo o o o o o o 11 42 Biacore 2000 107 K Kinetics analysis e o o o o o o o o o o o o o o o o o o o o o o o o o o o o o o 64 KINJECT eeso ooo 0 0 ooo oo oo oo oo o o o o o oo 42 L Linked Reaction amp o o o o o o o o o o o o o o o o o o o o o o oo o o o o o o o 60 gt o gt o o ooo oo ooo oo oo ooo oo oo oo o o o e o o o o o
86. Cycle OK Biacore92000 BI Ja wl blw Sequence EA DETECTION 2 1 3 1 4 1 F FLnw 20 p BASELINE baseline 5 INJECT Antibody 120 A INJECT END FN bound IF bound Subtracted RelResp lt 20 THEN EXIT CYCLE END IF Wie QUICKINJECT END EL 60 RESPONSE regeneration 5 Help Menu M If Then regeneration 100RU La 2 blw Sequence 37 DETECTION 2 1 3 1 4 1 amp FLOW 20 p BASELINE baseline 5 INJECT Antibody 120 INJECT END B 10 RESPONSE bound 5 IF bound Subtracted RelResp lt 20 THEN S EXIT CYCLE END IF QUICKINJECT 10mM Gly HCI pH2 0 20 QUICKINJECT END E B p RESPONSE regeneration 5 Help Menu Y VIT Biacore 2000 5 87 Ta If Then 3 IF RelResp For Report Point regeneration in Flow Cell Sampl
87. Date 02 Sep 1999 Analysis temp 25 0 C RESULTS A IFC Serial Sequential Fc 1 22165 19141 Fc 2 22469 21141 Fc 3 22501 21193 Fc 4 22461 21159 Fc 1 2 3 4 22439 Leaks 34 B Refractometer test 15 Sucrose Baseline level Fc 1 22209 15246 Fc 2 22510 15334 Fc 3 22538 x 15435 Fc 4 22499 5 15390 Average 22439 22000 23000 15351 Stdev 154 200 82 300 C Mixing Mix 1 Mix 2 Average Difference Dilution factor D Noise Short term Longterm Slope Fc 1 lt 0 3RU Fc 1 0 01 lt 0 05 RU s Fc 2 0 3 RU Fc 2 0 01 lt 0 05 RU s Fc 3 0 3 RU Fc 3 0 00 lt 0 05 RU S check check Biacore 2000 8 101 Bia Users
88. E 8 epos musa sr Is gt 2 Menu bar Biacore Toolbar O Sensorgram window 2 A erem Report point table Eventlog window A Status window RU Run Biacore 2000 1 3 1 2 1 2 1 Dock Tel Dock Lz Insert a new sensor chip Click Dock when ready After dacking you should run Prime from Working Tools
89. Enbe DET SAA EI MS RECAE D AVES ER 388g Don t Save GR PAJARES Me 11 S B8 y Save Ctrl CET Break Biacore92000 94 6 6 6 1 Standby JR COME 3 3 Standby Tools Start Standby 5 ul min 96 96 Standby 80 ml Standby MiliQO 2 OMilliQ97K C Prime
90. F2 50 50 mM NaOH Prime Before Run v Standby Flow After Run Menu lt Back Start Close Standby Flow After Run Standby 5ul min Prime Before Run Prime Start Biacore 2000 3 25 Ti BIACORE 3 xi 9 Do you want to save current wizard template Save Save As Don t Save Don t Save 11 Save Results As B 31 xl Save in Tanaka 3 ex En D test File name pH Scouting m Save as type Results blr X Cancel 7 Ya Sqave My Network P M y SER AND Ctrl CET Break
91. Fc 1 T iiS INJECT Cancel Wolume Contact Time More gt gt 1 Recovery Recovery Fraction Collection C Mone To Position rZal Recovery V nlume O Fractions 2 Extra Cleanup Extra Cleanup Options Predip Needle Position W Extra Cleanup Biacore 2000 4 47 4 2 4 1 Run gt Run Application Wizard gt Surface Preparation Start Ta Surface Preparation Surface Preparation Steps E X Surface Preparation Steps Immobilization pH Scouting Immobilization Immobilizatio
92. GE Healthcare 9 Biacore Biacore 2000 Ver 3 Instrument Handbook GE imagination at work 71 3202 31 1 ASS AAA A 1 AO Sr SUED col N AEA 1 1 1 1 gt 1 A A Nc ES 2 ASI AA MM Hed ee e d 3 Qo A 3 1 2 2 2 UP BERGE A etre 5 IP CAESA ID ol 7 ED A A E EA II 9 AE A 10 2 a O IA 11 2i a IE AR AN 11 A GE NN oo SSO 11 CNT O A a 14 E a A eu e ui esee Use MEM eR Es LENT LUE 15 2 205 55 CBS rettet ot e ote oe ceti fiti da fa dla ds EL LEER 16 PP 16 EE 17 AR 7 007 2D 7p EII AAE EE AEA 19 O eR A OA E decente du UM M M LEE MEL MN 21 CENE a a EQ Ei 27 3 153 d p eS CES A Soter tte ia 33 4 4 37 a A AS 40 4 2 ee 47 4 2 JIT IROTT EIE E ee Ree eed Eus 52 4 3 RM 56 4 3 1 ae 56 ME IJA A 60 Biacore 2000 5 ska EE sneen DU UTERE 64 5 1 kk 64 5 2 ll 69 E LN EE O PES PP oO E A O 74 IN DL ERR ERREUR 76 5 3 2
93. Level Baseline Level Response Level Baseline Level Response Level Biacore92000 56 4 A TOS ER 4 3 RINREPUAADO fc 9 0D SER ZR OD LAO 4 3 1 Mass Transfer rebinding
94. bles Unknown1 Unknown2 Unknown3 Unknown4 Unknowns Unknown1 Unknown2 Repl Analyte Conc Next gt Biacore 2000 Her lt gek New gt cos 2 Stqndqrd1 1 Standard1 2 ug ml _ 1 100 unknown 1 2 5 93 Next gt Standby Flow After Run Prime Before Run Start y Ei BIACORE E x 32 Do you want to save current wizard template Save Save As Don t Save
95. centration Series 2 xl Use Flow Cell s A Using 1 as reference Flow Rate 20 l min Stabilization Time fo min Injection Time A min Dissociation Time 3 min Run Order As Entered y Entered ala Repl Concentration pg ml vjjnM gt 150000 20 133 150000 10 150000 150000 150000 lt Back Next gt Close 20ul min 2 3min 2 3min Do Sorted Low to High As Entered Analyte Name MW Repl Concentration 5 Biacore92000 66 5 1 0
96. ch Grade 15 Ss IC IPR Ce S Certified Grade 5 Sensor Chip CM4 Sensor chip CM5 Sensor Chip CM3 Sensor chip CM5 Sensor Chip C1 Sensor chip CM3 Uis Sensor Chip SA E e AS AD a AAT T FARAS 1 CDNA A7 FPSS EAF 57 ETE TB Sensor Chip NTA NIA
97. e Biacore92000 HI A 44 4 ALE DS FE 22 2 10 10 30 Apply to All Curves Binding of DSA 100ug ml to immobilized Asialofetuin Ce 10000 0 Fr APR G Time Window AbsHesp 5D Slope Baseline HelHesp ld assay 305 50 221851 2 0 0 05 Yes O Baseline 2 assay 306 50 13243 5b 015 007 Yes Baseline 3 assay 30 7 50 13193 0 014 0 05 Yes Baseline 4 assay 238 50 13424 3 009 HE Yes Baseline 1 assay 170 5 50 267955 4 6927 36 993 No bound 2 assay 1706 50 13255 5 043 0 21 Ma bound 4 assay 1538 50 13422 5 0 20 004 Ma bound Status window D 38 View gt Reference Line
98. e 1 2 Biacore 2000 Sanitize 1 FC Sanitize BIA maintenace kit BlAdisinfectant solution R MilliQ zur 1 5 ml BlAdisinfectant solution BA 20 ml MiiQO Era o MilliQ97K 15 ml amp 2 7 7 Zr ootuelce amp c set13 Do 38 Sanitize O SANBA TITZ 2 Sanitize MiliOQ9 Pime 1 2 7 2 Amine Coupling HsA 1Dug mi 1200 1400 1600 1500
99. e lt C Comparison Is Greater Than y _ Cancel Help Constant Y Value f od Use Additional Condition THEN Command Block v IF RelResp For Report Point regeneration In Flow Cell Sample Comparison ls Greater Than Value Constant 100 THEN Command Block OK 52 w2 biw Sequence EA DETECTION 2 1 3 1 4 1 amp Flnw 20 E BASELINE baseline 5 A INJECT Antibody 120 INJECT END EL 10 RESPONSE bound 5 IF bound Subtracted RelResp lt 20 THEN A EXIT CYCLE END IF QUICKINJECT 10mM Gly HCI pH2 0 20 QUICKINJECT END A il PL 60 RESPONSE regeneration 5 IF regeneration S ample RelResp gt 100 THEN ENDIF END IF Als OKk Ta w2 blw Sequence B i BASELINE baseline 5 INJECT Antibody 120 A s INJECT END 10 RESPONSE bound 5 IF bound Subtracted RelResp lt 20 THEN EXIT CYCLE E END IF QUICKINJECT 10mM Gly HCI pH2 0 20 dr QUICKINJECT END B S RESPONSE regeneration 5 7 IF regeneration S ample BelBesp gt 100 THEN QUICKINJECT 1UmM NaOH 10 A C QUICKINJECT END 2 Emo F Help Menu M Biacore92000 88 5
100. ea Comcel so o E Fe1224 mm lt E ns TE e 3 Fc 4 Biacore 2000 4 41 HA Flow El Set Flow 3E Flow EM ul rain Cancel OK Ka kg 20 50ul min y Ctrl Break Y 20 View Plot Single 1 Curve Sensorgram Fo 1 mOUVOLb qm du 21e RIS s View Plot Overlay 97NC 0 9 View Plot Curve Classes Curves Ce O 7090374 Le Command gt Inject
101. eb Menu lt Back Hext gt Close Next gt Biacore 2000 54 4 HENX Tj surface Preparation Prepare Run THERMO A right hand rack position Position Min Vol pl Content 51 360 Antigen F3 300 SHmi ha Prime Before Run M Standby Flow After Run Help Menu Back Start Close Standby Flow After Run Prime Before Run Start CES BIACORE e Do you want to save current wizard template Save Save As Don t Save i 2 9 0 e N Dont Save 11 Save Results As Save in a Tanaka B3 e E3 D test abd File name Perfor mance test Save as type Results bir y Cancel Sqve y My Network P Ctrl
102. et os Biacore 2000 A CA 3 1 2 Run gt Run Application Wizard Fd BIACORE 3000 Control Software on COMI File Edt View Command Ma Tools Window Help A Hun Sensoargram ae HAZE k EE i dis Run Application Wizard Run Method Cata Collection Rate Ti Run Application Wizard Surface Freparahion Start Analyte Recovery Binding Analysis L ur se Template kinetic Analusis z Customized Application i in Cancel The Surface preparation application wizard guides vau through the essential steps in preparing the sensor surface for analysis including immobilization pH scouting ligand immobilization regeneration scouting and surface performance test Surface Preparation Start y Ja Surface Preparation Surface Preparation Steps E X Surface Preparation Steps Immobilization pH Scouting Immobilization NOMEN Immobilization in Surface Prep Regeneration Scouting Surface Performance Test Guides you through the steps in immobilizing ligand or capturing molecule on the sensor chip surface Help 4 Back Nest Llose Immobilization Next gt y Biacore92000 28 3 33 Surface Preparation Imm
103. generation 5 Help Menu bd Ba Nest Close Cycle wl blw Cycles Run Order As Entered v lv Use Separate Vials for Different Solution Variables E No1 QuM 1 No1 5 3uM 1 No1 12 5uM 1 No1 25uM 1 No1 5Q0uM 1 No1 100uM 1 No1 12 5uM 1 No2 QuM 1 No2 6 3uM 1 No2 12 5uM 1 No2 25uM 1 No2 50uM 1 No2 100uM 1 No2 12 5uM Menu lt Back Next gt Close Biacore 2000 5 75 30 roo km m in 36 Np s erc FU SIN 2 LIA X AE E Bita s gt YN J A Xx B1 285 0231 E DT AITZ KDE s X B i A al
104. n 10 mM Acetate pH 4 00 220 50 mM NaOH THERMO C y THERMO_A eeee Any changes you make to the rack settings here will not take effect until you run the wizard Help Menu lt Back Nest gt Close Menu gt Template Save As Oe OO e o nosnssssos e o nos ssssos cons sens 0600000000 O O O Next gt i 33 ph4 Prepare Run EE THERMO_A right hand rack position Position Volume pl Content A1 50 Ligand in 10mM Acetate pH 6 00 A2 50 Ligand in 10 mM Acetate pH 5 50 A3 50 Ligand in 10 mM Acetate pH 5 00 A4 5 Ligand in 10 mM Acetate pH 4 50 AS 50 Ligand in 10 mM Acetate pH 4 00
105. n Method Dissociation in Buffer Single Injection 1 Two Injections 2 2 Solution Injection Time Next gt Biacore 2000 5 67 Ti Kinetics Rack Positions tt 390_0 ug ml D nM Rabbit lnG 90 20 pg ml 133 nM Rabbit IgG 90 10 pg ml 66 7 nM Rabbit IgG 90 5 pg ml 33 3 nM Rabbit IgG 90 5 pg ml 33 3 nM Rabbit IgG 90 2 5 pg ml 16 7 nM Rabbit IgG THERMO C y THERMO_A OOOO0lesooo 99 0 OOOOl es5s5s5d eo ee 2 30 ecco 30 E EE NE eocool 00000 S080 0 0 Make sure that the correct rack s are installed in the autosampler when you start the run Help Menu lt Back Next gt Close Next gt y Ta Kinetics Prepare Run THERMO 4A right hand rack position Position Min Vol pl Content A1 90 O pel 0 nM Rabbit IgG 22 390 20 Id 133 nM Rabbit IgG A3 90 10 ginl 66 7 nM Rabbit IG 24 390 5 g m 33 3 nM Rabbit IgG AS 9 5 g m 33 3 nM Rabbit IgG AB 390 2 5 pon 16 7 nM Rabbit IG AT 90 1 25 pg mnl 8 33 nM Rabbit IgG 28
106. n in Surface Prep Hegeneration Scoutinr Surface Performance Test The Regeneration scouting wizard guides pon through the determination of suitable regeneration conditions Help Back Mest gt Cloze Regeneration Scouting Next gt sa Surface Preparation Analyte Settings El ES Analyte Settings Mame DSA Injection Time E mir Flow Rate 20 l min Aun Settings E Fiowcel A Help Back Next gt Cloze Name e p e Injection Time 2 5mim Flow Rate 10 20ul min Use Flowcell Next gt Biacore92000 48 4 a Ta Surface Preparation Rack Positions THERMO C THERMO A y OOIOIBSSSSCO LEE NEN CIT MPO ee ee 9 WE NE ooooo 0 OOOO 000000 EE A E S06 kooe Make sure that the correct rack s are installed in the autosampler when you start the run Help Menu lt Back Next gt Close Jy ue ECY Fe Not i Hi Surface Preparation Prepare Run 1x THERMO_C left hand rack position Position Volume ul Content A1 75 DSA Prime Before Run v Standby Flow After Run Help Menu lt Back Start Close
107. nked Reactions Analyses the dissociation phase of the interaction after varying lengths of time at steady state and presents the results as an overlay plot aligned ta the start af dissociation _ Heb S Hp Mei gt Cloze Linked Reaction Next gt y S Kinetics Assay Principle Fx Assay Principle Binding Using Capturing Molecule sptorina Barameters Hame Ligand Elow Rate 5 p rir Injection Time E min Stabilization Time 110 min Help Back Hest gt Close Direct Binding Binding Using Capturing Molecul Next gt Biacore 2000 4 61 Fi Kinetics Linked Reactions Contro BA Use Flow Cellls 1 Analyte Hame ET pg ml T SD Back Hest gt Cloze Use Flow Cell s Analyte Name PITA MW OO SE Next gt S Kinetics Regeneration Fx Regeneration M ethad Regeneration Flow Rate Flow Rate 30
108. nt Solution Variables j Rep Antibody Antibody2 b 1 Menu lt Back Next gt Close Antibody1 Antibody2 83 Biacore92000 84 5 C Program Files BIACORE2000 Guide Methods Wizard Templates V Aldehyde Coupling blu UT CD 7 Concentration Analysis bin 2 HPA Ohp Oomlgbl HPA P Ligand Thiol Couplingbln P POE Manual Ex Inhibition assayblw HERIR Manual Ex Consecutive Regeneration blw AUD E 4 Manual Ex3 jue Controlbl Moped sec MNA DM Manual Exd Positive Control every fifth cyclebln 2 NTA Chip Couplingbln A UE QUE REUS 22 NTA Chip Preparingbln NTA 2 Sh Chip Preparingblw SA Surface Thiol Coupling tly Run gt Run Application Wizard 7 J v 72 3 5 Customized Application Open template Y Open
109. nu lt Back Start Close Standby Flow After Run Prime Before Run Start y Cha BIACORE E xi 2 Do pou want to save current wizard template Save Save As Dont Save 9 0 0 E Dont Save 11 Biacore 2000 4 63 ES test File name Save as type Results blr E Cancel My Network P Save y Ctrl ET Break Ja 380602b Results Response Li 300 1000 1500 000 300 Concentration Series Baseline Level Mass Transfer Linked Reactions 11
110. nw 10 B BASELINE baseline 5 INJECT nalyte 50 d p INJECT END B P RESPONSE bound 5 INJECT 10mM Gly HCI pH2 0 10 LE lr INJECT END B A RESPONSE regeneration 5 Help Menu M FLOW 10 S El ES OR E Kewword mmm Cancel Help Analyte Keyword OK MEN eles maa Et e e TEMP 25 o B 2 E KEYWORD Analyte i Conc El UDS Ds lx Cancel Help E Keyword Conc Biacore92000 BI mr Ti Customized Sequence p p EB A L el 10 BASELINE baseline 5 INJECT Analyte 50 INJECT END 20 RESPONSE bound 5 INJECT 10mM Gly HCI pH2 0 10 INJECT END 60 RESPONSE regeneration 5 Help Menu bd ach Nest Close Next gt Run Order As Entered 1 Standard2 1 Standard3 1 Standard4 1 Standard5 1 Standardb 1 Standard 1 Standard2 1 Standard3 1 Standard4 1 Standard5 1 Standardb 1 Unknawn1 1 Unknown2 1 Unknown3 1 Unknown4 1 Unknowns 1 Unknawn1 1 Unknown2 Ti Customized Cycles x w Use Separate Vials for Different Solution Varia
111. o 4 84 M Mass Transfer e o o o o o o gt o gt o gt o gt o o o o o o o o o o o o oo o o o o o o o o 18 56 N Normalize e o 0 o o o o o eo o2 o o o o o 0 9 e e o eo o 10 NTA ooo ooo oo ooo e e e e e oo e ooo oo e oo 4 84 P Prime e e 1 0 o c c co cm c c c c c o c c c ooo ooo ooo o 5 6 94 96 97 Q QUICKINJECT e o o o o o o o o o o o o o o o o o o o oo o o o o o o o o o o 42 R Regeneration Scouting e o o o gt o gt gt o gt o o o o o 9 9 o o o o o o o o o o o o o o o o o 47 Rinse e e e e e gt e o c c o o c5 o o ooo ooo oo oo oo o 96 S SA 4 84 Sanitize e o o o o o o o o o o o o o o o o o o o o o o o o oo o o o o o o o o 97 Specify Flow Rate and Injection Time e e e eee o 0 0 0 28 U Unclogging e o o o o o o o o o o o o o o o o o o o o o o oo o o o o o o o o 97 Undock e o o o o o o gt o o gt o o gt o o gt o o o o o o o gt o o gt o o gt o o o o o o o o o o o 95 WI Wizard Template e o o o o gt o o gt o o gt o o gt o o gt o o gt o o o o o o o o o gt o o gt o o o o o 84 Biacore92000 www gelifesciences co jp e maill Web 2009 GE
112. obilization Kd E3 Sensor Chip Immobilization Method mine Coupling Help lt Back Nest gt Close Sensor Chip CM5 Immobilization Method Amine Coupling Next gt Immobilization Procedure C Aim for Immobilized Level Help Back Mest Close Aim for Immobilized Level 3 1 3 289 Specify Flow Rate and Injection Time Specify Flow Rate and Injection Time Next gt 12 NHS 5ul min 7 Eu ORIS LE 7 O 5ul min 7 Biacore 2000 3 29 Plow l min Back Henst gt Cloze Ligand Name Injection Time 23 RAND 7min Flow MIR 10ul min Next gt 13 O
113. termining kinetic parameters for an interaction helps you to design and perform crucial control experiments and gives immediate Feedback in the form af apparent rate constante based on a 1 1 interaction model usted pag E E EN EE RENE Kinetic Analysis Start y E kinetics Analysis Steps Kinetic Analysis Steps M Concentration Series E Control Experimente Be Mass Transfer Ee 27 Linked Reactions Performs analysis with a series af analyte concentrations to determine kinetic parameters Help 4 Back Hest gt Cloze Concentration Series Next gt U Biacore 2000 Direct Binding 5 65 C Kinetics Assay Principle Assay Principle Binding Using Capturing Molecule mihi Barameters Mame Ligand Elow Rate 5 ulmin Injection Time E mim Stabilization Time 10 mir lt Back Mest 5 Cloze Binding Using Capturing Molecule Direct Binding Next gt Use Flow Cell s Flow Rate Injection Time Dissociation Time Stabilization Time Run Order Random Ta Kinetics Con
114. ume pl Content C1 115 EDC Coupling Solution C2 115 NHS Coupling Solution C3 Empty vial for mixing minimum capacity 200 yl C4 210 Antibody C5 75 Ethanolamine C10 50 50 mM NaOH Prime Before Run v Standby Flow amp fter Run Help Menu lt Back Start Close Standby Flow After Run Prime Before Run Start i Ti BIACORE E El 2 Do pou want to save current wizard template Save Save As Dont Save 0 0 Mind Dont Save 11 Biacore 2000 3 35 Save Results As My Computer Es 5 inj My Network P Save y Ctrl Break ES 18 xi al EEepmielw s e ey mes zuo sl anti alb f ug ml target 1000 RU 34000 32000
115. y HCI pH2 0 Yar by Cycle Volume 20 uj F Vary by Cycle Options Predip Needle Extra Cleanup Injection Mode QUICKINJECT Solution Vary by Cycle Volume OK 2 7 95 Biacore92000 sB 79 D J Customized Sequence ARA ES DETECTION 2 1 3 1 4 1 F FLOW 20 A INJECT Antibody 120 g INJECT END OUICKINJECT 1 mM Gl HE pH 20 p GUICKINJECT END 2 INJECT Antibody 120 gA INJECT END e QUICKIMJECT 10mh Gly HCI pH2 0 20 g QUICKINJECT END 1 e AND EE Eon as Report Point i x v E Time hn 3 C Phal Injection Start _ Cancel window 9 s Cancel ld baseline 0 0 Help Iv Baseline din 10 Before ld Baseline OK Ti Customized Sequence F FLnw 20 B i BASELINE baseline 5 INJECT Antibody 120 el l INJECT END QUOICAINJECT THmkM Gly HCI pH2 0 20 A es QUICEINJECT END Biacore 2000
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