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Biacore X100 Plus Package 取扱説明書

1. Biacore X100 Assay Workflow ProteinA_mlqG Assay Overview Type of assay Direct binding e Selected chip CM5 Protein Results References Overview Sensor Surface Preparation e 4 J amp B30 Find Immobilization pH Find Sample Conditions Bento find out Run Binding Analysis Assay zz Pun Biacore X100 Plus Package
2. Biacore X100 Plus Package S BE 2 CFCA 20C 324 3x10 1 fxnreixMw173 m s f ERK Trel 20C Mw Da D Biacore Biacore X100 Biacore Web Biacore X100
3. V8 Immobilization Results Ligand Response Final RUF Target Reached Targetlevel Amine ProteinA N A No Preconcentration binding is too fast Preconcentration binding is too fast Target Reached Preconcentration binding is too fast pH Preconcentration binding is too slow
4. MER IV ji L 5 1 Other options wizards V Open New Wizard Template Surface Preparation Name Modified Immobilization pH Scouting Modified Last Week Immobilization 1 zd yiii s iie Ia kinetics Multi LMw 7372009 521 10 PA ia Assay Conditions Sample ia Regeneration Scouting LJ Assay C3 Concentration Z For acetazolamide 5 30 2008 6 56 17 F zj Using calibration a Calibration free CFCA Modified March 2009 zi Kinetics Affinity E Binding Analysis Ia b amyloid SCK 090318 3 18 2009 12 03 42 ia Custom Assay Wizard Ia HMGB1 Inhibit GFP Immob 090316 3 17 2008 7 33 00 F Modified Two Weeks Ago Modified November 2008 2 Demo 200811111 11 11 2008 2 53 13 Ia 1107 11 6 2008 2 20 11 F E m Biacore Templates Open Custom Assay Wizard
5. 30 1 1 BUE pH RI NaCl lt 2M 10 mM Gly HCI gt pH 1 5 HCI lt 100 mM Phosphoric acid lt 100 mM Formic acid 20 96 10mM Gly NaOH lt pH 12 NaOH 100 mM Ethanolamine 100 mM
6. 1 2 1 2 Biacore X100 Plus Package 4 91 Immobilize ligand covalently using Sensor Chip CM5 Create Assay Workflow Binding Analysis Ligand details Preview of recommended Assay Workflow Ligand name Protein amp Sensor Surface Preparation Find Immobilization pH Ligand attachment approach Immobilize My ligand is another protein Recommended Q Capture using own antibody and Sensor Chip CM5 Assay Immobilize ligand covalently using Sensor Chip CM5 Find Sample Conditions FindRegeneration Conditions Assay overview RunBinding Analysis Assay Type of assay Direct binding analyte Selected chip CM5 Ligand attachment overview Preview of recommended Assay Workflow
7. RU 10 Binding to reference Stability baseline Biacore X100 Plus Package 6 141 6 6 1 Concentration Analysis
8. crude Bulk Effect
9. AbsResp Baseline RelResp Baseline RelResp Binding to reference Binding level I Report Point Table Workarea Biacore X100 Plus Package Evaluation Explorer 9 177 9 3 Evaluation Explorer Sensorgrum All sensorgrams amp Work area Sensorgram window 9 3 1 Sensorgram window Curve Name Fc 2 1 me Cycle Purpose lt Overlay gt Cycle Overlay E Curve Name Fc 2 side lt an v2U Bio20 t
10. Biacore X100 Plus Package C Rmax 100 RU Rmax x xS Do RU Do S 50 000 Da 1 000 RU
11. Quick Filter Advanced Filters Biacore X100 Plus Package 4 1 AE 1 3 Quick Filter E
12. Biacore X100 Plus Package 8 171 Dock Chip New chip New chip Chip type cm5 Chipid 070510957 Chip lot no optional Chip type C Maintenance Dock Chip 2U v 2U x3 Tools gt Prime Biacore X100 Evaluation Software Prime Undock Biacore X100 control software Exit Start All Programs Oracle Database 10g Express Edition Stop Database HP Solution Center Internet Internet Explorer P 3 E mail J outookExpress hec Biacore x100 con 29 software Biacore X100 Ev Software we AllPrograms P Biacore X100 Plus Package 172 8 HE 8 1 Stop Database D Administrator Al Programs Oracle Database 10g Express Edition Stop Database My Documents L My Recent Doc
13. 100 40 50 BIA maintenance Kit 65 Superclean Desorb and Sanitize Tools More Tools Service Tools Superclean Stqrt Superclean This procedure cleans the flow system Total run time is about 100 minutes NOTE Use the Maintenance Chip for this procedure The surface on other sensor chips may be damaged by the solutions used Put the pump inlet tubes in filtered deionized warm water 40 50 C Next gt Superclean Required solutions 1 acetic acid 0 2 M sodium bicarbonate B M guanidine HCl 10 mM HCI If small molecule assays are run exchange the last two wash solutions for 50 DMSO and 10 DMSO respectively Next gt Biacore X100 Plus Package 7 167 V Superclean Rack Positions i
14. CM CM l Haat e 0 5 pH pH Immobilization pH Scouting Immobilization pH Scouting pH
15. Target Reached Preconcentration binding is too slow pH Biacore X100 Plus Package 3 41 AE 3 6 1 Wizards Immobilization Other options A Wizards Immobilization New V Immobilization Setup 7 Chiptype Fow cell 1 Aim for immobilized leve Blank irr Fow cell 2 Blank irr Help Custom Methods immobilize fow cell1 Method cvs _ _ Capturing molecule
16. Residuals for a good fit Residuals for a poor fit RU u 08 100 200 300 400 Time Biacore X100 Plus Package 3 79 ka 1 Ms kd 1 s KD M Rmax RU Conc M tc Flow ul min kt RU Ms RI RU Chi RU U value 1 576E 6 0 003258 2 067E 9 40 00 1 641E 7 1 000E 9 30 00 5 10OE 7 0 1585 3 500E 9 0 4129 3 7 500E 9 0 1520 1 500E 8 0 5975 3 000E 8 0 1167 GO Residuals G Parameters bulk effect 3 17 82 Finish Evaluation Explorer
17. Sensor Chip Maintenance 25 C BIA maintenance Kit BR 1006 66 BIAdesorb solution 1 90 ml BIAdesorb solution 2 90 ml BIAtest solution 65 ml BlAdisinfectant solution conc 10 ml BlAnormalizing solution 30 ml Sensor Chip Maintenance 1 BIAdesorb solution1 4 Biacore X100 Plus Package 7 161 7 1 Tools Undock Chip Biacore X100 This will undock the sensor chip Undock Chip Sensor chip
18. ng ml ug ml ko ER o 1 10 10 ug ml Biacore X100 Plus Package H
19. DMSO METS DMSO DMSO DMSO UV spectrometry Dimethyl sulfoxide 276855 Aldrich Dimethyl sulfoxide D 1435 Sigma 1XPBS 100 ml 1X PBS 5 DMSO 400 ml lt gt 1X PBS 500 ml 50 ml 10X PBS 450 ml S 100 ml 1X PBS 400 ml mm 20 ml 2 100 DMSO 20 ml 1X PBS 5 DMSO 400ml 3 DMSO 2 3 1 2 2 1XPBS 10 ml 3 1XPBS 596 DMSO
20. CC C HE 10 1 SN E i Er 5s Bj e ld la DE Biacore X100 Plus Package 190 A Add Report point ese DR Ede e a b e ea e D ee dele D e te e A piedi 21 Administrators 185 187 188 Affinity oec dee a e RS ORAE EE 86 87 88 Aim for immobilized level 38 40 41 102 B Bar Chart iiiter ier erp E DAN UE EE HEAD UN dU E En eo ey n E NEED DRE RO Da oe CENE ANE 127 Baselirie oec rere tes re DENN DER REA NR E edi 20 21 51 112 121 176 178 Ie 20 Binding Analysis 89 141 Bivdlent Analyte 2 ERAS e AE ARR RE Rt EI EE 76 Blankiimmobilization EE EEEE tr ei reti i dre ed e i e EE 41 102 Bu lkiEffect i2 tae eet E SE G C Current Fits Custom Methods El Eh lt 1 42 103 Custom Report Points
21. IF _ pd 3 suatia 60 4383 TA N p E KF esso
22. Dock Chip New chip New chip Chip type CM5 Chipid 070510 1014 Chip lot no optional Chip type Maintenance Dock Chip Dock Standby Tools Prime Help Shutdown Biacore X100 Evaluation Software Prime Standby Biacore X100 Plus Package 162 7 7 2 Tools More Tools B Maintenance Tools Desorb Desorb and Sanitize B E Test Tools amp System Check and Pump Calibration S Service Tools Software Problem Report Flow System Wash qi Superdean qi Clean Needle and Needle Wash Station 3 Calibrate Needle qi Replace Needle 3 Replace Peristaltic Pump Tube Cs This applicationprovides astandardformulaforreporting software malfunction to Biacore Last run time N A Tools
23. RI Biacore X100 Plus Package 3 81 RI 0 Constant 1 ka Ka Rn lt Rmax RU Ra
24. Biacore X100 Plus Package 1 5 1 2 1 2 1 Dock Chip Dock Chip 9 New chip New chip Chip type CM5 Chipid 070510 0957 Chip lot no optional ONew chip Reuse chip 1 4 Chip type Chip id Chip lot no
25. 3 4 1 1 lt gt Other options Mqnual Run DOS Manual Run Flow e Elow rate ul min Flow path O A Flow path 1 O Flow path2 Reference subtraction Flow path 1 2 2 1 v 30 l min Flow path 1 2 Reference subtraction 2 1 Load Samples Start Sqave
26. File Save 2 3 File Print Printer Printer HP Deskjet 5400 Series Sensorgram O None Current Cycle v Wizard Template ORwe Wizard Results O All cycles File Properties C Include event log for cycles OK File Properties Wizard Template Wizard Results Sensorgram None Current Cycle Range All cycles Include event log for cycles Biacore X100 Plus Package 24 3 3 3 1 Create Assay Workflow D kinetics Affinity Create Assay Workflow Kinetics Affinity
27. i Add Report point E Edit Report point Add x Report Point f Time 291 0 s Wmdw 5 t Baseline Apply to all curves in this cycle Help Cancel d Baseline 0 Biacore X100 Plus Package 22 2 2 1 3 End manual run Usb Run Stop Run Biacore X100 This will end the manual run when the last queued command is completed OK Standby 2 1 4 Standby Tools Stop Standby Biacore X100 Plus Package 2 23 2 2
28. Final Bound amp NHS Final Bound Immobilization Result Close Close Immobilize Overview Results reference Biacore X100 Plus Package 3 37 Aim for immobi
29. Rmo Rr Biacore X100 Plus Package 82 3 Big 3 17 F5 Kinetics Affinity Fit Kinetics Create Add Fit Curent Fits Model 1 1 Binding TT Description Parameters Dee Components 500 0 500 Quality Control Repor Residuals Parameters Curve ka 1 Ms kd 1 s KD M Rmax RU Conc M tc Flow ul min kt RU Ms RI RU 1 576E 6 0 003258 2 067E 9 40 00 1 000E 9 30 00 5 100E 7 0 1585 3 500E 9 0 4129 7 500E 9 0 1520 1 500E 8 E E 0 5975 3 000E 8 0 1167 lt Back gn cace Finish Add Fit Model Fit F5 Kin
30. text Fi Edi Format Vew Heb ycle 1 Fc 2 1 x Cyclecl Fc 2 1 Y RPoint X RPoint Y cycle 2 Fc 2 1 X cycle 2_Fc 2 B 63 0 240234 3 0 121094 6 0 852539 3 8 78906e 3 62 0 858398 62 0 318359 9 o 61 0 766602 o 61 0 866211 61 0 341797 60 0 280273 0 0742188 6 0 175781 E 60 1 41992 9 2 49805 1 06348 59 2 625 182 59 2 7002 185 0 80957 58 0 859375 185 58 0 483398 57 2 37012 187 7 2 3305 187 1 4 57 2 78906 197 4 75195 197 1 8016 4 56738 19 2 30371 56 3 9707 200 3 00293 200 2 36328 5 2 95094 200 2 70313 55 1 29102 202 0 273438 0 111 E 54 0 389648 0 689453 53 0 367188 53 0 0517578 0 956055 52 0 807617 532 0 321289 1 17578 51 1 26563 50 1 27 31 0 595703 0 658203 50 0 750742 50 0 275391 0 147461 49 0 259766 49 0 261719 0 839844 48 1 08594 47 48 0 537109 0 734375 47 0 930664 47 0 260742 0 459961 46 0 632813 46 0 0761719 0 0761719 45 0 208008 45 0 353516 0 0605469 44 0 339844 44 0 151367 0 746094 43 1 03613 42 43 0 654297 0 875977 42 1 08398 41 42 0 371094 0 167960 41 0 219727 41 0 568359 0 049804 40 0 261719 40 0 0351563 0 666992 39 0 901367 39 0 667969 0 507813 38 0 69436 38 0152344 0 113281 37 0 271484 0 0234375 36 0 226563 0 151367
31. Next gt E Start Save 4 2 95 Biacore X100 Plus Package 4 95 PPP Run Stop Run Biacore X100 N This will stop the run Stop Run Run Stopped Finishing current cycle please wait Abort cycle by Ctrl H Break Standby Ctl Breqakl Biacore X100 2 You have aborted the run Do you want to wash the system Recommended Yes Biacore X100 Plus Package
32. Copy V8 Custom Methods Surface thiol aureus Command Solution Contact Time s Flow Rate l min 79 PRECONC Spedfied in Immobilization Setup EDCNHSINXECT EDC NHS 50 50 WASH Ethanolamine LJ LIGANDINJECT Specified in Immobiization Setup INJECT Ethanolamine Method Method name Edit V8 Custom Methods Surface thiol Method name Copy of Amine Command Solution Contact Time s Flow Rate ulmin 79 PRECONC Spedfied in Immobiization Setup EDONHSINJECT EDC NHS 50 50 WASH Ethanolamine L LIGANDINJECT Specified in Immobilization Setup INECT Ethanolamine OK Set up Method Biacore X100 Plus Package
33. Biacore X100 Plus Package 1 9 Tools Set Temperature os re Prime Shutdown Biacore X100 Evaluation Software Stop Standby Load Samples v Rack Illumination Dock Chip Undock Chip Set Temperature Change Password Preferences More Tools 4 C 40 C OK 10C Set Temperature Analysis temperature C temperature temperature 5C 30
34. Biacore X100 Plus Package 10 1 1 2 4 Load samples O1 rack locked rack locked Load Samples Rack Ejected Insert the rack and click OK Time until automatic standby 4 52 Rack Ejected OK 13 rack locked Tools Rack Illumination AE 1 9 15 position 1 position 15 Po
35. Immobilization Result Close Close pao Immobilize Overview Results reference Biacore X100 Plus Package 102 4 RSS ERONEER A212 S AE 4 5 ZL O 1 Wizards Immobilization Other options A Wizards Immobilization New V Immobilization Setup 7 Chiptype Fow cell 1 immobilize fow cell1 Method Capturing molecule Aim for immobilized leve ligand solution Target level RU Wash solution 50 mM NaO
36. 1 2 1 2 Biacore X100 Plus Package 26 3 Immobilize ligand covalently using Sensor Chip CM5 Create Assay Workflow Kinetics Affinity Ligand details Preview of recommended Assay Workflow Ligand name Protein amp My ligand is another protein Sensor Surface Preparation Ligand attachment approach Recommended O Capture using own antibody and Sensor Chip CM5 Immobilize ligand covalently using Sensor Chip CM5 Assay overview Type of assay analyte 4 ligand Selected chip Find Immobilization pH Immobilize Assay Find S ample Conditions Find Regeneration Conditions Run Kinetics Affiny Assay Direct binding CM5 Ligand attachment overview GIZ Preview of recommended Assay Workflow Continue Save Assay Workflow As Save in folder Folder Cancel B nakayama Users amp Training By Group1
37. Solvent correction Extrapolate amp F Solvent Correction Extrapolation Range Extrapolation Range x axis 1000 RU 10 OK 30 1 2000 1500 1000 500 0 500 1000 1500 2000 2500 Response Ref Ru Biacore X100 Plus Package 138 5 100 Evaluation Add plot wy cerunt Relative response T Plot name Bl MW correction Plot type Report Point vs Variable Axis setting Y Report Point binding Response type MW adjustedresponse X Variable Cycle number woree oo 9 C
38. Sample Response Dhi Baseline RU 1 Sample Response Baseline 2 B25 4 1 Baseline Sample Response 5 7 2 Sample Response Baseline 5 2 7 2 Biacore X100 Plus Package
39. 10 30 Biacore X100 Plus Package L W ii Biacore X100 DMSO DMSO 3 596 DMSO DMSO 10 3 50 DMSO DMSO DMSO DMSO
40. 3 8 46 22 lg VE TRAINING Biacore X100 Contro EA Fie Edit View Commands Run Ta Injection command VialAwell position Contact time s Minimum required volume in vial well for this injection 50 ul Vial well position 0 Inject Vial position 11 6 Minimum required volume in vial for this injection 100 ul Contact time 60 120 W IV i G OK Biacore X100 Plus Package 48 3 Cancel Inject
41. CFCA dIR aF f M k Conc Mw Dadl 100 ul min d R dt p E Response s8 8 B EB B B CFCA 4388 150 kDa 5 000 RU Mw km PH Con d R dt 2 5 100 l min km D 2 1 1
42. Regeneration solution1 2 Contact time 1 3 59 V8 Regeneration Scouting Rack Positions 105 10mM Gly HCl pH3 0 Regeneration 135 10mM Gly HCI pH2 0 Regeneration Full Deionized water Water Hee Menu v _Lead Samples lt Back Nen gese Rack Positions Next gt Start l Squve 3 2 30 TRAINING Biacore X100 Control Softwar EA fie Edit View Run Tools Help e Regeneration Scouting 5 11 2007 2 12 18 PM B v9 X oe RU PHI H ng 24800 OW 3 f i i E z FEIM R tin 0 50 Fc Time Window AbsResp SD LRSD 2 57 0 5 263764 033 019 2 230 5 26713 045 009 Temperature 25 00 ec E t EE TRAINING B
43. Ko Ko 1 5 uM 5 FARAI CIR THU BE Ko Ko Next gt Biacore X100 Plus Package 70 3 PE Kinetics Affinity Rack Positions Sample 150000 105 m IgG Sample 150000 105 m IgG Sample 150000 105 mIgG Sample 150000 105 m IgG Sample 150000 105 m IgG Sample 150000 1275 Buffer Startup 105 Buffer Startup 225 10mM Gly HCl pH2 0 Regeneration Full Deionized water Water 1 2 3 4 5 6 7 8 9 H gt lt Back Me tee Rack Positions Next gt Start Biacore X100 Changes detected
44. DMSO 10 RU Solvent correction 4 b Biacore X100 Plus Package N 3 100 RU 10 000 RU DMSO 30 000 RU
45. J Next gt EE Concentration Analysis Rack Positions Ear 125 m 19G Calibration 10 125 m IgG Calibration 5 125 m IgG Calibration 2 5 125 m IgG Calibration 1 25 125 m IgG Calibration 0 625 125 m IgG Calibration 0 313 125 m IgG Calibration 0 70 samplei Sample 70 sample2 Sample 70 sample3 Sample 290 buffer Startup 85S 10mM Gly HC Regeneration Full Deionized water Water Jil gt lt Back Net J Gee J Rack Positions Next gt Start Biacore X100 Plus Package 6 145 Biacore X100 Changes detected in wizard template Save As Don t Save Squve 6 1 BE 6 1
46. QCratio 0 13 lt 0 13 Biqcore X100 Plus Package K Calibration free concentration Select Samples Create Expand all cycles Show original sensorgrams KM iE aed factor ul min prel RU s prel Ceo 25 10 Fc 2 1 antibeta2micro Sample 500 0 873 11800 100 10 100 Fc 2 1 antibeta2micro sample 25 11800 10 100 10 View K Sample Sample Curve Fc 2 1 mwe Show blank subtracted data 1 362E 10 1 362E 10 RU Sample Fc 2 1 120 4 Time 6 Flow 10 ul min 7 Flow 100 plmin 10 Flow 10 plimin 11 Flow 100 plmin E Zoom lock Included blanks Advanced settings Include View 4 Sample mouse IgG 1 5400 Curve Fc 4 3 Cancel 157 Show blank subtracted data
47. 50 ml HBS EP 4 C 2 S Fe
48. Biacore X100 Plus Package New Cycle Yc Commands gt New Cycle New Cycle Flaw p Flow rate ulmin Flow path O E Flow path 1 O IE Flow path2 Reference subtraction Flowpat 12 Flow path OK End manual run jv E Run gt Stop Run Biacore X100 2 This will end the manual run when the last queued command is completed OK Standby Biacore X100 Plus Package 3 53 lt gt Assay Find Sample Conditions m Run to find out Enter known values Find Regeneration Conditions m Run to find out Enter known values Run Kinetics Afinity Assay rl F
49. admin Biacore X100 Control Software Manual Run 10 27 2008 2 14 43 PM Ez Bie gdt View Commands Run Tools Help Servke3 xd TO XB se ce smoesnrc2 cT WE D V Lock scale Mus Regeneration 12 60 jaselne_1 1 binding 1 stabity_1 F baseine 1 i 300 Time AbsResp Slope RelResp Baseine ld _Keywords in cyce 1 Value 219 36 00 00 Yes baseine_1 243978 11 67 24240 No bindna 1 244532 354 24856 No stabilty_1 206781 001 00 Yes baseine 206783 003 02 No binding 1 206837 007 48 No stability 1 Fn 30 Fon pan L2 Online COMI Temperature 25 00 c Running manual run Fc 1 Biacore X100 Plus Package 4 D 1 Regeneration Command iin Dd iL ss Regeneration Vial position 12 Contact ine a Minimum required volume in vial for this injection 70 pl
50. Run gt Stop Run t This will stop the run Help Stop Run Cancel Stop Run Finishing current cycle please wait Abort cycle by Ctrl Break Standby Ctrl Break Biacore X100 You have aborted the run Do you want to wash the system Recommended Yes Standby Biacore X100 Evaluation Software Biacore X100 Plus Package 146 6 admin Biacore X100 Evaluation Software Concentration Analysis 5 17 2007 4 24 19 PM Fie View Evaluation Tools Window Help S E ij Sensomram Plot A Kinetics Afinty Binding Analysis L Concentration Analysis Sensorgram Biacore X100 Plus Package 6 147 Biqcore X100 Evaluatio
51. Biacore X100 Plus Package 9 175 9 Biacore X100 Evaluation Software Biacore X100 Evaluation Software 9 1 Windows Start Programs All Programs Biacore Biacore X100 Evaluation Software 9 2 lt File Open OK Biacore X100 Plus Package 176 9 Evaluation explorer Evaluation Explorer Sensorgram I Plot Baseline Binding level Binding stability 38539 d dPPII Work area
52. pH 5 200 ug ml 10 mM Immobilization pH Scouting 50 mM NaOH Biacore X100 Plus Package G IV IV i
53. Ctrl oe Ove EADH 2 v amp alsicitind DRE Mapag Ctrl Biacore X100 Plus Package 178 9 9 3 2 Sensorgram window LL Iob v Tools Color By Sample
54. IE Kinetics Affinity Rack Positions 915 m IgG Sample 105 m IgG Sample 150000 105 m IgG Sample 150000 105 m IgG Sample 150000 105 m IgG Sample 150000 105 m IgG Sample 150000 1275 Buffer Startup 105 Buffer Startup 225 10mM Gly HCl pH2 0 Regeneration Full Deionized water Water E Menu lt Load Samples lt Back Next gt Close Menu Automatic Positioning Automatic Positioning Change the order in which the samples are positioned by ordering the regions The first region in the list is positioned first Regeneration El Brown Pooling Yes Pooling No OK Biacore X100 Plus Package 72 3 3 5 3 5 1 admin Biacore X100 Evaluation Softwar
55. Ko 10 0 30 60 l min 3 3 5 10
56. Ligand details My ligand is Ligand attachement qpproach d biomolecule with a tag My ligand is tagged with Sensor Chip SA Biotin CAPture kit 28 9242 33 GST GST capture kit BR 1002 23 his Sensor Chip NTA qnother tag an antibody My antibody is d mouse antibody Mouse Antibody Capture Kit BR 1008 38 d human antibody Human Antibody Capture Kit BR 1008 39 qnother antibody e another protein Immobilize
57. x ws IE E Curve Name Fc 22 6d cycle Pupose vete vli Cycle caveri we Took v RU Sensorgram D d am ia B a Bi 6100 da Bi ia Bi gy Repor 6050 id Report Point Table son sen 8 sample PU Saian K E Startup 5500 ESI E 5650 0 1200 1400 1800 Biacore X100 Plus Package 134 5 5 2 BE 5 1 Keyword Table Tools Keyword Table fF Keyword Table Cycle Cycle Purpose Sample Conc pg ml MW Da Ww v w w Evaluation Add Solvent correction Ei Solvent Correction Response Act Ref N Extrapolation Range Report Point Range 20 t t t T t 1 Nae
58. V Regeneration Scouting Save Settings First regeneration Xa den W 10mM Gly HCI pH2 0 30 Second regeneration Use these settings in the workflow steps Stabilization period Save Biacore X100 Plus Package 62 3 Assay Workflow ProteinA_m lgG Ligand AttachmentOverview Type of assay Direct binding Selected chip CM5 Ligand name ProteinA Edit Assay Workflow Sensor Surface Preparation Results reference Immobilization pH Scouti Em em Immobilization buffer 10 mM Acetate pH 5 S2007 10 14 37 AM All results 3 a Method in fc 2 Ami ine Pr 5 11 2007 Immobilized in fc 2 451 3 RU 11 50 13 AM Chip id 070511 0939 All results Results reference Assay Conditions Sample Find Sample Conditions DE 5A172007 12043 PM time 60s All results Run to ind out Find Regeneration Conditions 1 Regeneration solution 10mM G
59. Biacore L7IV28 ES 97 V283 RR dSORg 9 7 V288 05 S V Mad 34680 97112 ll Biacore X100 Plus Package B II 90
60. CM5 N g CM NHS N NHS S S QS S S
61. Biacore X100 Plus Package 38 3 Ligand 175 ul 11 mm 50 mM NaOH 70 ul 11 mm Ethanolamine 126 ul 11 mm EDC 85 HV 11mm NHS 85 pl 11mm Ze NHS EDC 11 mm EDC NHS Next gt Start i sqve ctr Break EE TRAINING e Biacore X100 Control Software Immobilization 5 11 2007 11 50 13 AM EA Fie Edt View Tools He UFA F Cycle EV Curve Sensorgram rc ProteinA 42000 l bilzed W gt Puiset 2 8 3 H 3 d
62. Continue Save Assay Workflow As Save in folder Folder B3 nakayama i Users Name Modified B Training ProteinA mlgG2 5 9 2007 2 26 05 PM Ea Group Z PretenAmlgG 5 9 2007 9 58 30 AM E muan 070509 5 9 2007 9 27 23 AM gt os B2 5 8 2007 2 31 33 PM dti 5 5 8 2007 2 23 32 PM Bg Shemy Yoshimi uc 4 5 8 2007 2 08 54 PM 1 By Software exercises Bg system check 5 8 2007 2 08 04 PM administrator Instrument Service amp ProteinA m IgG Save Biacore X100 Plus Package 92 4 T admin Biacore X100 Control Software Eile yiew Run Tools Window Help Services Assay Worktiw Proteina mlgG X AS Type of assay Direct binding 0 PR Ligand name Proteina ligand Selected chip CM5 Edit Assay Workflow Sensor Surface Preparation Overvi Results reference FindImmobilization pH
63. E admin Biacore X100 Control Software Manual Run 10 27 2008 2 14 43 PM EA Ele Edit View Commands Run Tools Help Service3 x6 FE oen one so TP E New Cice 302 P iniecit 120 V f Regeneration 1260 EE P binding 1 tabiity 1 beseine_1 AbsResp SD LRSD Slope RelResp Baseline ld Keywords in cycle 1 Value 29736 007 007 00 00 baseline 1 243976 2183 015 1167 24240 bindng 1 244592 663 013 354 24856 stabilty 1 2067931 005 005 001 00 baseline 1 208783 007 003 003 02 binding 1 208837 018 014 0 07 45 stabilty 1 UEXCTUM Onine comi Temperature 25 00 C Running manual run ji Fc 1 Biacore X100 Plus Package 50 3 IV i H Regeneration Command A amp 2U v 2U id Regeneration Vial position 12 Cnet ine 6 Minimum required volume in vial for this injection 70 pl
64. Find Sample Conditions Enter known values Next gt Biacore X100 Plus Package 4 119 V8 Regeneration Scouting Experimental Parameters Regeneration parameters Stabilization period Experimental design Number of conditions 2 Number of cycles for each condition El Settings Condition Regeneration solution Contact time s 10mM Gly HCI pH3 0 30 10mM Gy Hd pH2 0 Next Close Experimental Parameters Regeneration parameters Stabilization period Experimental design Lock Solutions 1 culo du des Ard ouin Solutions Contact time 1 Solutions Contact time
65. Next gt La basa bl ei la tn 7E06 09 SE 6 7 10 11 Eee owi DO 13520 10 te 8 9 12 13 Processing series 2 of 4 Abot remaining Iteration 48 Man relative change 0 1254 kd 0 002900 Accept randm row S0 ein 8 now On Pied Piw SO ite n2 few t ini Paed 12 Five 105i 85 Pow 00e Fited 13 fu 100m Biacore X100 Plus Package 158 6 Calibration free concentration Evaluation Result Create Expand all cycles re Dilution Meas Conc Calc Conc QCratio Temp Mw D Chiz SE Blank Fit Cre Ligand Sample factor M A M fit C Da mz s RU Meas Conc Cycles subtr status Fc 2 1 antibeta2micro Sample 500 7 5E 09 3 TE 06 0 933 25 11800 1 362E 10 0 043 3 6E 11 6 7 10 11 Yes Completed Fc 2 1 antibetaZmiro sample 100 3 8E 08 3 8E 06 0 961 25 11800 1 362210 0 068 41E 11 8 9 12 13 Yes Completed sample Fc 2 1 Zoom lock AM 8 Flow 10 ul min Fitted 8 Flow 10 plimin 9 Flow 100 plimin Fitted 9 Flow 10O UlImin 12 Flow 10 ulimin Fitted 12 Flow 10 Hlmin 13 Flow 100 HUImin Fitted 13 Flow 100 ul min 50
66. 3 BR 1000 12 3 BR 1000 14 1 BR 1003 99 3 3 3 BR 1005 39 BR 1005 41 BR 1005 40 DNA 3 BR 1000 32 1 BR 1003 98 3 BR 1000 30 1 BR 1004 06 3 BR 1005 43 1 BR 1005 58 His tag 3 1 BR 1000 34 BR 1004 07 BR 1005 42 Biacore X100 Plus Package 8 1 1 2 2 ZY YTRER KAFEE Tools gt Prime Help Shutdown Biacore X100 Evaluation Software Place a bottle with buffer on the left hand tray and insert the inlet tubes Start Prime Prime Priming please wait Time left 5 56 The Prime procedure is completed Close Standby AE 1 7 Prime
67. Next gt V Binding Analysis Samples sample1 sample2 sample3 sample4 sample5 sample6 sample7 sample8 sample9 sample10 405 10mM Gly HCl pH1 5 Full Deionized water Es Ce se Cue yen Ca Rack Positions Next gt Biacore X100 Plus Package 126 4 Start Sqve 4 2 95 Standby Standby 2 1 4 22 Biacore X100 Evaluation Software TRAINING Biacore X100 Evaluation Software Kinetics Affinity 5 14 2007 2 08 23 PM O X 2 Support Navigator Fie View Evaluation Tool
68. Binding Analysis Kinetics Affinity Overview IV iv P Other options Wizards Concentration Using Calibration V amp Concentration Analysis Injection Sequence Detection Flowcel EM v Injections in analysis cycle Fowcell1 Fowcell2 Cow Enhancement Regeneration Injection Sequence Detection Flow cell Chip Chip type Injections in analysis cycle Capture Enhancement lt Next gt Biacore X100 Plus Package
69. Reuse chip Reuse id Dock Chip O New chip Reuse chip Reuse chip Reuse CA Chip type CM5 Chip id 070426 1110 Chip lotno 1170296 Chipid Chiplotno 070426 1110 1170296 Chip IFC type CM5 IFC5 Immobilization Final Response Capturing date RU Molecule Ligand SEHE 4 26 2007 1450 3 Beta 2 Mab Immobilization 4 26 2007 11 20 56 AM Close Biacore X100 Plus Package 1 7 HE 1 6 Biacore X100 Sensor Chip CM5 certified Sensor Chip CM5 reserch grade Sensor Chip CM5 reserch grade Sensor Chip CM4 Sensor Chip CM3 Sensor Chip C1 Sensor Chip SA Sensor Chip SA Sensor Chip HPA Sensor Chip HPA Sensor Chip L1 Sensor Chip L1 Sensor Chip NTA Sensor Chip NTA Sensor Chip Au
70. Tools Report Point Id and Marker 9 3 3 Sensorgram window Libr Tools Sensorgram Adjustment E Adjust Sensorgram of O Report Point time 0 Injection Event time 0 X Adjustment Report point time 0 FT baseline X Adjustment C Off Report Point time 0 8 n Injection Event binding LE ime 0 stability Y Adjustment Report point response 0 FT baseline 9 j amp 2Uv2Uzs Biacore X100 Plus Package 9 179 9 3 4 amp Adjusted sensorgram Zoom Lock RU 1800 1600 4 1400 4 1200 4 10004 5 Beta2micro high2 amp a 8004 Beta2micro high1 p Beta2micro low2 600 Beta2micro low1 amp gt E 400 4 E
71. Blank 0 Blqnk Next gt Biacore X100 Plus Package Concentration CFCA Rack Positions E CNE NE ec MW Da 20 C Sample Yes 291 Sample Sample 11800 1 19E 10 291 sample Sample 11800 1 19E 10 162 buffer Startup 405 Glycine HCl pH2 5 Regeneration i Ful Deionized water Water Help Menu w LoadSamples lt Back Next gt Rack Positions Next gt e Start Sqve 6 1 145 Standby Standby 2 1 4 22 Biacore X100 Evaluation Software
72. V Regeneration Scouting Injection Sequence Detection Flow cell Injections in analysis cycle FowCelli HowcCell2 2 Next gt V8 Assay Conditions Sample System Preparation Prime Prime before run Conditioning Run conditioning cycle Prime before run Prime Next gt Injection Parameters Find Sample Conditions Find Sample Conditions Enter known values Next gt Biacore X100 Plus Package 58 3 V8 Regeneration Scouting Experimental Parameters Regeneration parameters Stabilization period Experimental design Number of conditions 2 Number of cycles for each condition E Settings Condition Regeneration solution Contact time s 10mM Gly HCI pH3 0 30 10mM Gy HGpH20
73. Samples MW Dq Da Biacore X100 Plus Package 3 65 I Kinetics Affinity Rack Positions 105 mouse coG Sample 150000 105 mouse IgG Sample 150000 195 mouse IgG Sample 150000 105 mouse IgG Sample 150000 105 mouse IgG Sample 150000 105 Buffer Startup 285 10mM Gly HCl pH2 0 Regeneration Full Deionized water Water 3 4 5 6 7 8 H Rack Positions Next gt WEE Start squve 3 2 30 Standby stqndby 2 1 4 22 Biacore X100 Evaluation Software S TRAINING Biacore X100 Evaluation So
74. d biomolecule with a tag My ligand is tagged with Sensor Chip SA Biotin CAPture kit 28 9242 33 GST GST capture kit BR 1002 23 his Sensor Chip NTA qnother tag an antibody My antibody is Mouse Antibody Capture Kit BR 1008 38 Human Antibody Capture Kit BR 1008 39 d mouse antibody d human antibody another antibody e another protein Immobilize
75. Biacore X100 Plus Package 84 3 HE 3 18 Current Fits Current Fits Delete OK Current Fits Biacore X100 Plus Package 3 85 3 5 2 admin Biacore X100 Evaluation Software AAL_LNFP1 File View Evaluation Tools Window Help S Sensogam Plot uj BarChat Aj Kinetics Affinity L Concentration Analysis E All sensorgrams x Remove Edit Curve Name Fc 21 Cycle Purpose lt Overay gt E E Cycle lt Overay gt Sensorgram E Sensorgram All sensorgrams Plot i Baseline i Binding level ia Bindin
76. Mw km Bivalent Analyte CFCA Christensen Anal Biochemistry 1997 249 p 153 Sigmundsson K e A Biochemistry 2002 41 p 8263 Biacore X100 Plus Package R CFCA 438 5 000Da ka 107 gt gt 5X104M 1s 1 150 kDa Cl amp 5 000 RU CFCA 0 5 50 nM 1 ug ml 280 nm 10 4
77. EDC NHS Next gt We Start Sqve Ctrl Breakl 4 2 95 I admin Biacore X100 Control Software Immobilization 10 27 2008 11 56 13 AM EA BE Edi view Run Tools Help Service3 CA IEA IERA RU ProteinA 45000 3 B B2 8 E E X aa ass t t 500 2000 indow AbsResp SD Slope RelResp Baseline Il Keywords in cycle 1 Value 204634 005 0 01 00 Yes Baseline Chip CM5 206225 016 006 1591 No ContactTime 420 219185 058 031 14551 No Ligand FlowRate 5 219564 011 006 14930 No Immobilized Ligand Protein Method Amine j Procedure TimeAndFlow Temperatur Sensor chip CMS Running standby remaining time 6 9 days Standby Standby 2 1 4 22 Immobilization Results
78. Oracle Database 10g Express Edition 23 Administrators 2 GB Biacore X100 Control Software Tools Preferences Database Backup Preferences Schedue C Monday Tuesday wednesday 7 Thursday Fida T Saturday Sunday Sattine 23270 m Backup location Backup folder CXFrogram Filas Basora Biaon 100 Dalabasek 4 O Enable second copy Folder NumEei of saved backups 7 2 X Riestoie Available backups for restore based on backup leider backup craaed 4 1 2007 11 00 55 PM Schedule Start time Backup location Backup folder C ProgramFiles Biacore BiacoreX100 Database O Enable second copy
79. Desorb Standby Standby 3 4 Close Biacore X100 Plus Package 164 7 7 2 2 Desorb and Sanitize 1 1 1 20C Tools More Tools Maintenance Tools Desorb and Sanitize Start 2 Y v Desorb and Sanitize This procedure removes adsorbed material and disinfects the flow system This procedure includes 4 steps You will need to change solutions between steps The total run time is about one hour followed by a recommended standby time of 3 4 hours Next gt Desorb and Sanitize NOTE Use the Maintenance Chip for this procedure The surface on other sensor chips may be damaged by the solutions used Do not run this procedure below 20 C Do not abort this procedure after it is started Next gt Desorb and Sanitize Require
80. i B Immobilize 2Uv2UzsS Immobilization Setup Chiptype CM5 Prime before run Prime Flow cell 1 E Immabilize fow cell 1 Method Aim for immobilized level Ligand solution ee Amine Flow cell 2 Immobiize fowcal2 Method X Amine Q Aim for immobilized level Ligand solution Protein in 10 mM Acetate pH 5 r Specify contact time Contact time 1 2 7 Next gt V8 Immobilization Rack Positions Type 126 Ethanolamine Immobilization 75 Protein Immobilization 85 EDC Mix Fc2 85 NHS Mix Fc 2 Empty EDC NHS min capacity 130H Mix Fc 2 Full Deionized water Water Help Menu Load Samples lt Back ii Next gt Close jil
81. 103 107 1 Ms ka 10 5 0 5 1 s ka Rmax 4 R RI RI Residuals Y
82. Next gt Save Settings V Immobilization pH Scouting Save Settings M T S Sp Use these settings in the workflow steps 7 IYI e z Buffer Commen Buffer name pH 10 mM Acetate tais 5 5 10 mM Acetate pH 5 10 mM Acetate pH24 5 e essen B5 RELE pH Save Biacore X100 Plus Package 98 4 IE admin Biacore X100 Control Software Eile view Run Tools Help Service3 usPO X548 Assay Workflow ProteinA_mlgG Assay Overview Type of assay Direct binding anae Selected chip CM5 Edit Assay Workflow Ligand name Protein ligand Sensor Surface Preparation Overview Results reference x Immobilization pH Scouting Findimmobilization pH Immobilzation buffer 10 mM Acetate pH 5 RE 2 All results L Run to find out Enter known values Immobilize zi Run Overview Results reference Find Sample Conditions Bun to find out Enter known values Find Regeneration ondmons Bun to find out Enter known values
83. 10 ml Biacore X100 Plus Package M Frostell Karlsson A et al 2000 Biosensor analysis of the interaction between immobilized human serum albumin and drug compounds for prediction of human serum albumin binding levels J Med Chem 43 10 1986 92 SPR RD 100 RU DMSO
84. 64 3 V Kinetics Affinity Injection Parameters Sample Contact time 120 s Dissociation time 12 First regeneration Solution 10mM Gly HCI pH2 0 Contact time 30 s Stabilization period Injection Parameters Next gt IE Kinetics Affinity Samples Sample id 9 0 pg ml mouse IgG 150000 0 mouse IgG 150000 1 25 833E3 mouse IgG 150000 2 5 16 7E 3 mouse IgG 150000 5 333E3 mouse IgG 150000 10 66 7E 3 mouse IgG 150000 20 0 133 mouse IgG 150000 0 CES mouse IgG 15000 5 33 363 Sample Saqmple id Concentration 2 5 2 1 2 IV ii 1
85. Biacore X100 Plus Package 3 45 sad B ore 00 Co ol So are a alR 0 008 EA Ele gdt View Commands Run Tool Help Service3 Bx xd r0 X E45 oen gwe seeeeawren 3 iue 41 Help m Lock scale 25000 4 OE a M Y li New 20000 15000 10000 4 8 E f a 5000 4 04 5000 4 10000 t t t t t t t 1 0 5 10 15 20 25 30 3 40 45 50 Time Fc Time Window AbsHesp SD LRSD Slope HeHesp Baseine ld Keywords in cycle 1 Value Flow 30 Flow path 1 2 Online comt Temperature 25 00 C Running manual run 1 2 2 1 AE 3 7 1 View Show Only Current Curve View Show All Curves View Show Curves of Same Type Biacore X100 Plus Package
86. 10 HBS EP 200 ml BR 1001 88 0 01M HEPES 0 15 M NaCl 3 mM EDTA 0 005 v v Surfactant P 20 pH7 4 HBS P 200 ml BR 1003 68 0 01 M HEPES 0 15 M NaCI 0 005 96 Surfactant P 20 pH7 4 HBS N 200 ml BR 1003 69 0 01 M HEPES 0 15 M NaCl pH7 4 RREZEVE REMOS SE TARI DEZIR 0 22 um Biacore X100 Plus Package 1 3 1 1 4 Start Programs Biacore BIACORE X100 Control Software qdmin administrator 10 T TRAINING Biacore X100 Control Software Hi ues ros eb Sono Analysis Other Options Filtered on Folder nakayama Modified Yesterday keewcyAmmwy 5 5 2007 2 706 PM 5 9 2007 3 02 58 PM PAmgez Regeneration Scouting 5 9 2007 1 30 08 PM I Assay Conditions Sampie 5 3 2007 10645 PM 5 9 2007 1 28 58 PM l reseten 5 9 2007 11
87. s OK 111 ad B 00 Co ol So are a a 0 008 EA Ele Edit view Commands Run Tools Help Service3 Fe FEAS Cycle 1 Cuve Sensorgram Fc 2 gj i Hel RU Lock scale 25000 4 cp ge le KE ycle 3 E 3 24500 4 Y UM MM 24000 4 28500 23000 j 2250 4 ij 3 p 5 22000 4 W 21500 3 21000 20500 20000 4 a p t t at t 1 0 100 200 300 400 500 600 700 800 900 1000 Time s Fc Time Window AbsResp SD LRSD Slope HeHesp Baselne Id Keywords in cycle 1 Value 2 1760 5 219 38 007 007 00 Q0 Yes baseine 1 2 3010 5 249976 2183 015 1167 24240 No binding 1 2 360 5 244532 663 013 354 24858 No stabilly 1 l Flow 30 Flow path 1 2 He onine coMl Temperature 25 00 C Running manual run View Reference Line 1 Biacore X100 Plus Package 112 4 ad B 00 Control Software Manua 0 008 Ea Ele Edt View Commands Run Tools Help Services 8x FE B oe ce scone E ig Referencel
88. 4 2 95 V8 TRAINING e Biacore X100 Control Software Regeneration Scouting 5 11 2007 2 12 18 PM EA fie Edit View Run Tools Help B v9 X oe w OX dw 3 f i i E z FEIM R tin 24800 0 50 Fc Time Window AbsResp SD LRSD 2 57 0 5 263764 033 019 2 230 5 26713 045 009 Temperature 25 00 ec E t EE TRAINING Biacore condiyion sample 19 SF TRAINING Biacore 1 Biacore X100 Plus Package 4 121 Regeneration Scouting Results Sensorgrams A Sample Response Baseline Result Standby Trend Chart 9 7 Response Baseline Display 1st cycle 1 Conditions
89. 2020 7020 MO OO 23 r Mes ID E T EEEE EEEE 23 3 es 24 3 1 E B A a a D a 24 3 2 pH a eie rn ee denen te deatur 28 ee 34 2350 lt 44 3 A L 20200 44 3 4 1 1 eteeenenttttttnn 44 2 63 3 4 2 5 Jp Wt 2 Ele d SEL edet eere eee eel 67 3 5 SU NET cn c uU ME ct M iE eie cues ME 72 3 54 Jur 2m be NIE edm iem 72 225 2 85 4 LODE 10D D S A cd SC I dii 89 4 1 c 5 JEDE BI A E EESE 89 4 2 U HY KREIRAO pH RIR ase ebbe ac eee reb 93 3 99 4 4 ww 105 4 4 1 aa A Vy AE E S E 105 4 4 2 41 trot rtetetestettoest tiet tpat 114 4 5 2 AA 124 5 128 128 52572 134 6 MENTE orn 141 6 1 141 141 6 2 zz 151 Biacore X100 Plus Package 49 QU Io gis quo ndr E i A 160 X30 TROU AE i mtd adag aa Aseo E 161 Q1 AIT S ADET tdt pid eL CAD DM E NRI 162 LETT
90. 12 1 Open New Wizard Template S74 Surface Preparation R z Workflow Z Assay Conditions Capture fg Assay Conditions Sample REiegeneratien Scoutipa E Concentration zj Using calibration Kinetics Affinity Binding Analysis zj Custom Assay Wizard Biacore X100 Plus Package 2 13 2 2 1 Other Options Manual Run TRAINING Biacore X100 Control Software Create Assay Workflow Kinelics Affnity Other Options 2 wizaras Binding Analys
91. Biacore X100 Plus Package 4 115 V8 Assay Conditions Sample System Preparation Prime Prime before run Conditioning Run conditioning cycle Prime before run Prime Next gt V8 Assay Conditions Sample Injection Parameters M mm v Sample 1 Conc pg ml 50 mouse IgG Sample 50 mouse IgG Sample 50 mouse IgG Sample 50 mouse IgG Sample Full Deionized water Water Rack Positions Next gt Biacore X100 Plus Package WEN Statt l Save 4 2 95 EE TRAINING Biacore X100 Control Software Assay Conditions Sample 5 11 2007
92. F Kinetics Affinity Fit Affinity Create Steady State Affinity Report Parameters KD M 4 222E 4 G Report Kp M Rnmox RU REREXER Biacore X100 Plus Package 88 3 offset RU bulk effect Chi RU G Parameters Finish Evaluation Explorer Kinetics Affinity jd Kinetics Affinity Biacore X100 Plus Package 4 89 4 4 1 Create Assay Workflow D la Binding Analysis
93. Assay Find Sample Conditions m Run to find out Enter known values Find Regeneration Conditions m Run to find out Enter known values Run Kinetics Afinity Assay ma ze Find Sample Conditions Run to find out F T Enter known values 2 Uv 2L C Assay Conditions Sample Injection Sequence 12 Reference subtraction Dore IP I Ee Y 1 2 ane 5 Find Regeneration Step Next gt
94. NHs NHS pH8 5 pH pH pH5 Immobilization pH Scouting RU 100 ng ml Immobilization pH Scouting Next gt Save Settings Immobilization pH Scouting Save Settings ie Save Biacore X100 Plus Package I admin Biacore X100 Control Software File View Run Tools Window Help Service3 Arika ER 2 3 3
95. E Susie Roque 4 285A 50 A o EILAN 9530 975 30 eum S Cd fees OD eR kda B M kr M por
96. Find Sample Conditions Bunto find out Enter known values F 2 Find Regeneration Condiions Bunto find out Enter known values RunkKineticsZAffinty Assay 5 E Temperature 25 00 C Sensor chip CMS Running standby remaining time 7 0 days Run to find out L amp Run 2 5 Overview Results reference Enter known values D DREADS DE Overview Biacore X100 Plus Package 28 3 3 2 pH pH I i F Sensor Surface Preparation Sensor Surface Preparation Find Immobilization pH Run to find out Enter known val
97. kak 1 Globol Fitting R f Ks Ka Rmax C oe SS Req Req vsC C Req 07D0vHF25 SINIEE Ka Ko
98. Calibration free concentration Included blanks Include Cycles Curve Flow bl min ivi 4 Fc 4 3 5 i 5 Fc 43 100 i 1 10 F 3 Em 1 m3 100 Zoom lock P jj ij ka Wi Mh TURN i di TMAN Include DF IY Include OK 6 6 AE 6 6 CFCA CFCA C Initial rate RU s 20 3 15 RU s lt 0 3 RU s gt 15 RU s
99. 46 3 AE 3 8 5 10 30 ul min 72 53 amp 1R Detection1 2 Support Navigator Standby 49 ld 4 Biacore X100 Plus Package 3 47 77 241 F omn
100. 5 Biacore X100 Plus Package P W iv A B B A RU A RU vs C A AU Slop A L Slop vs C 2 B w A 7 oi A i P d Conc ug ml A
101. Biacore X100 Plus Package 7 163 7 2 1 Desorb 1 1 25 20C Tools More Tools Maintenance Tools gt Desorb Start This procedure removes adsorbed material from the flow system Total run time is about 25 minutes NOTE Use the Maintenance Chip for this procedure The surface on other sensor chips may be damaged by the solutions used Do not run this procedure below 20 C Use deionized water as running buffer Next gt Required solutions from Maintenance Kit BlAdesorb solution 1 BlAdesorb solution 2 BlAdesorb solution 1 BlAdesorb solution 2 1 5 m 1000 15 mm 4 ml H2O Start
102. Detection1 2 Support Navigator Standby 49 ld X Biacore X100 Plus Package 108 4 4 7 in
103. sensor chip T E3 Ili Dock Chip Dock Chip Biacore X100 Control Software Biacore X100 Plus Package 6 1 Dock D I sensor chip Standby Standby 7 HE 1 4 Dock Dock Undock sensor chip sensor chip HE 1 5
104. 0 5 2 pH pH 3 5 10 mM pH SA 100 ng ml 10 mM Borate 1 M NaCI pH 8 5 NHs pH 8 5 DMSO Instrument handbook Biacore X100 Plus Package F Ill ii pH CM5
105. 200ul 11 mm 20C 1 4C 200 pl Biacore X100 Plus Package E 1 BSA BE 10 mM 10 mM Borate 1 M NaCl pH 8 5 5 200 ug ml 10 mM pH
106. Run Binding Analysis Assay Run n Binding Analysis Injection Sequence Detection Flow cell 1 2 Injections in analysis cycle Fowcel1 FHowcCell2 r woes Injection Sequence Next gt I Binding Analysis System Preparation Prime Prime before run Prime Conditioning Run conditioning cycle Startup Run startup cycles Solution Buffer Numberofcycles 3 Next gt Biacore X100 Plus Package 4 125 I Binding Analysis Injection Parameters First sample Contact time 60 s Dissociation time s First regeneration Solution 10mM Gly HC pH1 5 Contact time 30 s Stabilization period 0 s Injection Parameters
107. Web 2010 GE GE T 169 0073 3 25 1 TEL 03 5331 9336 FAX 03 5331 9370 e mail Tech JP ge com 10 02 2 EP
108. b d 200 4 5 J 2 5 B o 200 50 0 50 100 150 200 250 300 350 400 s Time 0 baseline Sensorgram window wv Caption Cut Scale Copy Graph Export Curves Gridlines Legend Cut RU Adjusted sensorgram smrTeck 400 350 3 b 300 B f 250 H 200 Beta2micro high2 n i Beta2micro high1 H Beta2microow2 150 eg 5 Beta2micro low1 EN 100 prs 50 e j E 3 Bj 0 50 50 0 50 100 150 200 250 300 350 400 s Time 0 baseline Biacore X100 Plus Package 180 9 9 4 Sensorgram window v Caption Undo Cut Scale Copy Graph Export Curves Gridlines Legend Scale X Scale Y Scale Auto Auto C Logarithmic C Logarithmic Mag May Min Max X Scale Y Scale C Auto Auto C Logarithmic C Logarithmic in 5 Min
109. Next gt E start Save 3 2 30 Biacore X100 Plus Package 30 3 BE 3 2 Run gt Stop Run Biacore X100 M This will stop the run Help Stop Run Cancel Stop Run Run Stopped Finishing current cycle please wait Abort cycle by Ctrl Break Standby Ctrl Breakl Biacore X100 2 You have aborted the run Do you want to wash the system Recommended Yes Biacore X100 Plus Package 3
110. 122 4 Gsensorgrams Display Cycles Next gt Close V Regeneration Scouting Save Settings First regeneration Ep EE yO Lee lt Second regeneration Solution Use these settings in the workflow steps Stabilization period Save Biacore X100 Plus Package 4 123 F8 TRAINING Biacore X100 Control Software Assay Workflow ProteinA m IgG Ligand Attachment Overview Type of assay Direct binding Selected chip CM5
111. A A A A B B A j am A TED A A Biacore X100 Plus Package Q CFCA Calibration Free Concentration Analysis CFCA
112. Number of cycles Next gt V Kinetics Affinity Injection Parameters Sample Contact time s Dissociation time s Next gt Biacore X100 Plus Package DE Kinetics Affinity Samples Samples Sample id Dilution 150000 1500 lt Back Next gt Close Sample id MW Da Do Highest Concentration 5 Dilution Concl1 Conc 5 0 1 0
113. 7 169 V8 System Check and Pump Calibration Rack Positions BlAtest Solution 1 5 ml 123 ul 1 H20 4 ml Sstart By system check ne lied Dad Te System Check and Pump Calibration 5 10 2007 11 08 51 AM CES CER Name Save System Check and Pump Calibration System Check and Pump Calibration Date 10 May 2007 250 C System Check and Pump Calibration 5 10 2007 11 08 51 AM Peristaltic Pump Calibration Response Injections System Check and Pump Calibration Name Date Created By Biacore X100 Control Software Version 10RC2 lmetrumantit 42412 Pass Dzemesfied 88887 C pfild Fail Fail Biacore X100 Plus Package
114. 74 3 FS Kinetics Affinity Select Curves Create Curves Sample 7489 Temperature Contact Time Include Cycle Sample C rir 30 180 0 E Zoom lock Show concentration series Showblanks 7 Show average blank s sample 3 12 AE 3 12 Include Curves Temperature Curve Fc 2 1 Flow Contact Time ul min S 3 180 0 Ligand cOooooooooo Next
115. 8 Number of conditions 7 Number of cycles for each conditions 5 Settings Regeneration solution Contact timels 2 Regeneration solution1 2 Contact times 1 2 Next gt Biacore X100 Plus Package 120 4 ES Regeneration Scouting Rack Positions 105 10mM Gly HCl pH3 0 Regeneration 135 10mM Gly HCI pH2 0 Regeneration Full Deionized water Water Hee Menu v Load Samples Rack Positions Next gt Start Save
116. Next Close Experimental Parameters Regeneration parameters Stabilization period Experimental design Lock Number of conditions Number of cycles for each conditions Settings Regeneration solution Contact time s 2 2 Next gt Biacore X100 Plus Package Solutions 1 Contact time Solutions Contact time 1 Solutions Contact time B 7 5
117. x Remove Edi Sensorgram jm All sensorgrams a Plot a Baseline E Report Point T able m Report Point Table 75 Kinetics Affinity Z489 Al Kinetics Affinity Biacore X100 Plus Package 80 3 HE 3 16 RI 0IRU Residual Residuals Y Residuals for a poor fit RU Residuals for a qood fit U 084 054 24d 02 0 J gt ce 221 5 0 4 4
118. 1 20 000 Da Rmax 20 000 x 1 000 50 000 x 1 400 RU 10 000 RU RUvsC Ko RU RU 200x1 Sx RU 1 000 x 1 S x S 50 kDa 100 kDa 1 200 x 1 1 x 50 000 100 000 100RU
119. 31 birding start Em mm fE My Pictures LI FUE 1 Standby standby 2 1 4 22 Results Immobilization pH Scouting Results ProteinA 10 mM Acetate 5 5 10 mM Acetate 5 10 mM Acetate 4 5 Relative Response 10 mM Acetate 4 Y baseline 2 10 mM Acetate 5 5 ProteinA 2 10 mM Acetate 5 ProteinA 2 10 mM Acetate 45 ProteinA 2 10 mM Acetate i 4 ProteinA pH pH5 0 Biacore X100 Plus Package 32 3 dig 3 3 pH pH BARTIA pHi pH
120. Biacore Templates Biacore Templates Biacore X100 Plus Package 5 129 Select Biacore Template ta tE zx Binding analysis with solvent correction a Off ROBES A 2 J Ia Binding analysis CFCA template C Z Mut cycle nece ih solvent we m Z Single cycle kinetics with solvent correction Single cycle kinetics gs 3 Open V Custom Assay Wizard System Preparation Chip Detection Flow cell Reference subtraction Purpose Evaluation purpose Binding Analysis Prime Conditioning Prime before run Hun conditi Binding Analysis Concentration analysis using calibration Calibration free concentration analysis Evaluation purpose
121. Custom Report Points Add Add Custom Report Point Report point ld Window 5 6 Position the report point 10 seconds C Calculate response relative to report point Cycles Apply To Selected Cycle Purpose Cycle Purpose O Startup Apply To Selected Cycles Calibration Sample ld Cycles HE 6 4 Calibration Table Exclude Cycle Import Cycle Biacore X100 Plus Package 150 6 Samples KS Concentration Analysis Create 22 samplei Response Calc pg ml o l i 26 p
122. Report Point stability 6 3 Response Type Relative Response Sope Relative Response Fitting Function 4 Parameter Linear 4 Parameter Calibration Table Curve Cycles Conc ResponselRU Calc Conc CV Calc Conc Biacore X100 Plus Package 6 149 AE 6 3 Report Point Stobility 20
123. 10 l min 100 ul min CFCA I9G CFCA IgG IgM CFCA O AEL mol mzs x km Y D xf km 0 98 x 0 3 xh xw xl D fo h w
124. 170 8 8 8 1 2 Standby 7 Standby 7 8 1 1 Standby standby 7 7 200 ml online COM1 Temperature 25 00 C Sensor chip Ch5 Running standby remaining time 7 0 days 8 1 2 Tools Undock Chip Biacore X100 This will undock the sensor chip Undock Chip Sensor chip
125. 173 E 8 2 Oracle Database Stop Database Oracle Database Oracle Database DOS amp Stop Database iC NoraclexeNappNoracle sproduct 108 2 0 XservuerMBIN net stop OracleService E The OrkacleSeruice amp E service is not started El More help is available by typing NET HELPMSG 3521 iC NoraclexeNappNoracle product 18 2 80 serverNBIN Oracle Database 6 Stop Database Biacore X100 Plus Package 174 8 8 2 4C
126. Number of saved backups 7 Biqcore X100 Plus Package 10 189 CC Restore Restore OK
127. 1 000 x 1 1 x 50 000 100 000 500 RU 100 500RU Biacore X100 Plus Package D M i N g CM NHS N NHS CON COO COO CONHCH CH OH NHS RU BR 1000 50 EDC N ethyI N 3 dimethylaminopropyl carbodiimide hydrochloride NHS N hydroxysuccinimide 1M ethanolamine hydrochloride pH 8 5 EDC NHS 10 ml
128. Load Samples f exis Rack Unlocked Rack Unlocked Insert the rack and click OK OK Injection command A Vialwell position I 8 Lo Contact time s Minimum required volume in vial well for this injection 50 ul Biacore X100 Plus Package 3 49 admin c Biacore X100 Control Software Manual Run 10 27 2008 2 14 43 PM EA Ele gdt view Commands Run Tools Help Service3 Subtracted Fc 24 le New Cyce 302 YP Inect1112 s 2500 4 i baseline 1 200 250 Time Fc Time Window AbsResp ReResp Baseline ld Keywords in cycle11 Value 21 176 0 5 12945 00 Yes baseline 1 21 3010 5 37187 24242 No binding 1 21 3160 37755 2481 0 No stabilty_1 Fn 30 Fon pa DZ Onine COMI Temperature 25 00 C Running manual run Fc 2 1
129. Req Cx Rmax C Ko Req RU Conc M Biacore X100 Plus Package K Ko 1 10 10 5 0 1 2 n 2 1 Ko Ko 1 10 10 5 0 2 1 nM 1 nM 5 5
130. 4 105 4 4 1 Other options 2 Manual Run H l KE Manual Run Flow ge Flow rate pl min Flow path O E Flow path 1 O E Flow path 2 Reference subtraction G B Flow path 1 2 2 1 v 30 ul min Flow path 1 2 Reference subtraction 2 1 Load Samples Start Sqave Biacore X100 Plus Package 106 4 3 ad B ore 00 Co ol So are a al R
131. Next gt Biacore X100 Plus Package KineticsZAffinity Kinetics heterogeneous analyte 130 5 DE Custom Assay Wizard Cycle Definition ee gindng Analysis Solvent correction Commands in Binding Analysis Settings for Sample 1 Available Selected Solution varies by cycle Sample Enhancement Regeneration Contact time 60 s 7 varies by cycle Solvent correction Dissociation time B s varies by cycle Flow rate 10 pl min varies by cycle Flow path Both v Extra wash after injection with Remove Evaluation variables Cycle types Binding analysis Commands in Add Settings for Contact time 30 60 s Dissociation time 30 60 s Flowrate 10
132. Rack Positions Biacore X100 Plus Package 3 35 Ethanolamine 126 ul 11 mm Ligand 75 pl 11 mm EDC 85 pl 11 mm NHS 85 pl 11 mm Ze NHS EDC 11 mm EDC NHS Next gt Start Sqve Ctrl Breakl I admin Biacore X100 Control Software Immobilization 10 27 2008 11 56 13 AM EA BE Edi view Run Tools Help Service3 CA IEA IERA RU 45000 4 ProteinA 3 B B2 8 E E X aa ass t t 500 2000 indow AbsResp SD Slope RelResp Baseline
133. Sensor Surface Preparation Sensor Surface Preparation Find Immobilization pH a Run to find out Enter known values Immobelize Find Immobilization pH Run to find out Enter known values V Immobilization pH Scouting Setup pH 10 mM Acetate 10 mM Acetate 10 mM Acetate pH Next gt Biacore X100 Plus Package 94 4 PE Immobilization pH Scouting Injection Parameters Surface regeneration This surface wash will be run once at the end of each cycle 50 mM NaOH Close 45 ProteinA Sample 10 mM Acetate 5 45 ProteinA 1 Sample 10 mM Acetate 4 5 45 ProteinA Sample 10 mM Acetate 4 135 50mM NaOH Regeneration Full Deionized water Water Rack Positions
134. 06 T t t 6 100 0 100 200 300 400 500 100 200 300 400 s Time s Time Chi amp U value 15 25 SE Standard error Parameters SE SE 10 RI
135. S OK ad B 00 Co ol So are a a 0 008 EA Ele Edt View Commands Run Tools Help Service3 2x Ar iia ME EN Cyce 1 Cuve Sensorgram Fc 2 sj amp i Hel RU Lock scale 25000 i jc 2 i E HE 24500 4 A f Dr M 24000 1 28500 23000 4 g 5 225004 S E fi 22000 4 L tl s 21500 T 21000 20500 20000 4 t A paa t t A4 AA aa a 1 0 100 200 300 400 500 600 700 800 900 1000 Time s Fc Time Window Abshesp SD LRSD Slope HeHesp Baseine Id Keywords in cycle 1 Value 2 1760 5 219 38 007 007 00 Q0 Yes baseine 1 2 3010 5 249976 2183 015 1167 24240 No binding 1 2 3160 5 244592 663 013 354 24856 No stabilly 1 el Flow 30 Flowpath 1 2 Hen onine com remperature 25 00 C Running manual run View Reference Line 1 Biacore X100 Plus Package 3 51 ad Biacore X100 Control Software Manua 0 27 2008 EA tie Edt View Commands Run Tools Help Services Sx xd rO5tb5E5 583 Bee 1 Curve Sensorgram Fc 2 3 Es DH Referenceline X cal 156 0 5
136. 5 Fc 2 1 500 30 60 0 60 1 6 Fc 2 1 250 30 60 1 60 0 7 Fc 2 1 125 30 60 0 60 0 8 Fc 2 1 62 5 30 60 0 60 1 9 Fc 2 1 31 25 30 60 0 60 0 10 Fc 2 1 16 30 60 0 60 1 11 Fc 2 1 8 30 60 0 60 0 12 Fc 2 1 4 30 60 1 60 0 13 Fc 2 1 2 30 60 0 60 1 Remove Selection Undo Blank Subtracted Sensorgrams E Zoom lock p imm ey ire hr erg Re PU A m ch tm 1 100 120 140 Back ird Affinity gt lod Kinetics gt Cancel J 0 Affinity gt F5 Kinetics Affinity Select Affinity Data Create Calculate response at position 4 seconds before injection stop with window 5 seconds 3e 4 Concentration RU Req RU Req Biacore X100 Plus Package Next gt F Kinetics Affinity Fit Affinity Create 3 87 Steady State Affinity Fit Affinity Model Steady State Affinity Fit
137. Create Assay Workflow Binding Analysis Preview of recommended Ass Ligand details Ligand name ay Workflow My ligand is x Ligand details VUE My ligand is a with a tag BET den pecora ERR something else Binding Analysis Create Assay Workflow Binding Analysis Ligand details Preview of recommended Assay Workflow Ligand name Protein amp My ligand is another protein Nj Ligand attachment approach Recommended L Q Capture using own antibody and Sensor Chip CM5 Immobilize ligand covalently using Sensor Chip CM5 Continue Cancel Biacore X100 Plus Package 90 4 AE 4 1
138. BIAtest solution HBS EP 10X BR 1008 26 Sensor chip CM5 HBS EP CM5 Dock L Prime Tools gt More Tools Tools Maintenance Tools Desorb A Desorb and Sanitize Test Tools System Check and Pump Calibration S Service Tools EY Software Problem Report 3 qi Flow System Wash qi Superdean qi Clean Needle and Needle Wash Station qi Calibrate Needle Replace Needle Replace Peristaltic Pump Tube neni re Crinan 4 This procedure checks for leakage and tests pumps injections and noise The peristaltic pump is calibrated Total run time is about 35 minutes NOTE This procedure should be run at 25 C with a new Sensor Chip CM5 Running buffer should be HBS EP Last run time N A Test Tools gt System Check and Pump Calibration Start System Check and Pump Calibration Ensure that you have HBS EP as running buffer the analysis temperature is 25 C and a new Sensor Chip CM5 is docked HBS EP HBS EP 10X 10 Next gt Biacore X100 Plus Package
139. Ligand details Ligand name My ligand is a biomolecule with a tag an antibod another protein a nucleic acid a vesicle liposome something else Kinetics Affinity Create Assay Workflow Kinetics Affinity Choose ligand attachment approach Preview of recommended Assay Workflow Ligand details Ligand name maeasawaases LU EE Een Immobilize ligand covalently using Sensor Chip CM5 Biacore X100 Plus Package 3 25 AE 3 1 Ligand details My ligand is Ligand attachement qpproach
140. OR C E OFF
141. http www biacore com jp lifesciences products systems_overview x100 service index html Biacore X100 Biacore X100 Biacore FORGOT FASSWORD gt LOGIN User ID Password LOGIN Biacore X100 Plus Package T Biacore X100 Biacnre Biacore Biacore X10I amp 10 E Diffusion Coefficient Calculator Tool Fanrt
142. s lt Back Finish Cancel Meas Conc M Calc Conc M Meas Conc QC ratio fit QC EE Chi RU SE Meas Conc Meas Conc 6 7 Finish Evaluation Explorer E Concentration amp nalysis LA CalibrationFreeConc 1 Biacore X100 Plus Package 6 159 HE 6 7 CFCA D Chiz 5
143. Biacore X100 Plus Package 4 101 V Immobilization Results Chip CM5 Ligand Response Response 1 How cell Procedure Method 1 Bound RU Final RU Time and flow Amine ProteinA i 1296 0 1493 0 Response Bound Response Final RU AE 4 4 EEEE Response Bound Final 2 Bound Final NHS EDC Final Bound NHS Final Bound
144. DMSO DMSO 1 1 500 RU DMSO Biacore X100 Evaluation Software DMSO DMSO 19 x y GO M
145. pH pH5 0 Biacore X100 Plus Package 4 97 HE 4 3 pH pH pH4 pH NHS NHS pH8 5 Un pH pH5 Immobilization pH Scouting RU 100 ug ml RE BE Immobilization pH Scouting pH
146. 142 6 V Concentration Analysis System Preparation Prime Prime before run Conditioning Run conditioning cycle Startup Run startup cycles Solution Number of cycles Prime Prime before run Prime Startup Run startup cycles 3 Next gt V8 Concentration Analysis Injection Parameters Sample Contact time s First regeneration Solution 10mM Giy HCI pH1 5 Contact time s Stabilization period 60 s Injection Parameters Sample Contact time First regeneration Solution Contact time Stabilization period Biacore X100 Plus Package 6 143 Next gt V amp Concentration Analysis Calibration Curve Calibration Curve Analyte name Cali
147. 20C 20C Biacore X100 Plus Package V Biacore X100 Plus Package 1 1 1 1 1 1 1 1 gt gt gt gt biacore Windows LED power XIJIL temperature IO 7 zj7 9 l d Eli ss 1 1 2 2 1 2
148. 400 May OK Biacore X100 Plus Package 9 181 Legend Position O Hidden zs O Top Right O Bottom Right OK Hidden OK Gridlines Gridlines X Axis C Major Gridlines C Minor Gridlines Y Axis C Major Gridlines C Minor Gridlines 2 Major Gridlines Minor Gridlines OK Biacore X100 Plus Package 182 9 9 5 Sensorgram window Copy Graph Biac
149. 96 4 EE admin Biacore X100 Control Software Immobilization pH Scouting 10 27 2008 11 14 12 AM u E TT 4 150 ime Window AbsResp SD LRSD Slope RelResp Baseine ld Keywordsin cycle 1 Value 5 206349 010 010 402 00 Yes baseine DyciePupose Sample 118 0 5 184441 079 011 042 21908 No binding Running immobilization pH scouting 1 Standby Stqandby 2 1 4 22 Results V8 Immobilization pH Scouting Results ProteinA cpndng 10 mM Acetate 5 5 bindin binding 10 mM Acetate 5 10 mM Acetate 4 5 Relative Response 10 mM Acetate 4 140 160 180 200 E RelResp RU 2 10 mM Acetate 5 5 Protein 2190 8 2 10 mM Acetate 5 Protein 1440 1 2 10mM Acetate 4 5 ProteinA 2876 9 2 10 mM Acetate 4 Protein 6134 3 Ce CER c C Cas
150. Biacore X100 Plus Package O DMSO 5 DMSO DMSO DMSO 1x PBS no DMSO 4 6 DMSO 496 DMSO A F v 2785 1x 9 600 ul 100 96 DMSO 400 ul 10 000 ul 696 DMSO A F v Z8 1x 9 400 yl 100 96 DMSO 600 ul 10 000 ul 4 6 96 DMSO 8 DMSO 496 DMSO 100 200 300 400 500 600 700 696 DMSO 700 600 500 400 300 200 100 700 700 700 700 700 700 700 700 pl _DMSO uM Kp 1 10 10
151. M OK Biacore X100 Plus Package 2 17 AE 2 1 5 10 30 ul min 72 53 amp 1R Detection1 2 Support Navigator Standby 49 ld 4
152. CD TEL 03 5331 9336 xt T C UN USE ERAS e 9 00 17 30 0 TEL 03 5331 9315 FAX 03 5331 9349 TEL 06 6305 4707 FAX 06 6305 3599 Q TEL 03 5331 9336 0 2 O Q FAX 03 5331 9370 e mail Tech JP ge com WWW delifesciences COJD e mail
153. Next gt Start Biacore X100 t Y Changes detected in wizard template Help Save Save As Dont Save J Cancel Sqve As Dont Save Sqave Ctrl Breakl l Standby XYY KOZE Surface thiol Method name Biacore X100 Plus Package 3 43 Methods Copy Custom Methods Surface thiol Method name Copy of Amine Command p PRE CONC
154. SE standard error 10 0 05 5 ug ml HE 6 8 Evaluation Explorer g Kinetics Affinity P IB2 u elobulin Backspace c9 Kinetics Affinity 10C Biacore X100 Plus Package 160 7 2X 7 Tools More Tools Maintenance Tools
155. anae Ligand name 9 ligand Selected chip Direct binding Protein CM5 Edit Assay Workflow Sensor Surface Preparation Find Immobilization pH Overview B Run to find out l Vresciizaten bufer 10 rl Acetate pH 5 Enter known values Immobilize Metetinto 2 ame Imwewipedie o Z 4513 RU 1 Cap Oo Results reference FindSample Conditions Overview B Run to find out Enter known values Find Regeneration Conditions Bun to find out Enter known values RunKinetics Affiniy Assay Results reference Help Online COM1 Temperature 25 00 9C Sensor chip CM5 Running standby remaining time 7 0 days Immobilize Overview Results reference Biacore X100 Plus Package 40 3 Aim for immobilized level NHS
156. ligand solution Target level RU Wash solution 50 mM NaOH immobilize fow cel2 Method Capturing molecule el ligand solution Target level RU Wash solution 50 mM NaOH Chip type Method Surface thiol OAim for immobilized level Target level Wash solution Specify contact time OBlank immobilization Next gt 50 mM NaOH Biacore X100 Plus Package 42 3 Rack Position
157. test2 2 23 2009 6 57 01 PM administrator Assay Workflow test2 Biacore X100 Plus Package 184 9 File gt Export gt Result To Excel Evaluation Explorer xls Microsoft Excel xls Microsoft Excel 2 Name Sample 3 4 Sample Sample 5 Temperatu 25 6 Curve Fc 2 1 8 9 Model Bivalent Analyte Description 10 11 Curve ka1 1 Ms kdi 1 s ka2 1 RUskd2 1 s Rmax RU Conc M tr Flow ul mkt RU MS RI RU Chi RU2 U value 12 28071 45 0 019845 0 000287 0 06355 15688 84 3764036 2643 192 N A 13 Cycle 2 2 4E 09 30 11695736 2 114536 14 1 2bE 08 9 153725 15 6E 08 58 73426 16 3E 07 219 0913 17 1 5E 06 274 0177 18 19 20 9 7 File Save ASs Folder i nakayama Name Modified Crea Type Keval 5 23 2007 2 40 09 PM Training Eva
158. Biacore X100 Plus Package 18 2 Cancel L Inject Load Samples Load Samples Rack Ejected Insert the rack and click OK Time until automatic standby 4 45 OK Injection command amp Z Vialwell position I 8 Lo Contact time s Minimum required volume in vial well for this injection 50 ul Biacore X100 Plus Package 2 19 un Todi Heb HEE g Folre oe E eE d T Lock seale T E HET PAESE LT pm p 50000 asooo aaa E 35000 30008 25000 Xg BB 5 Bou T ii our 2m am su E 1000 1 oo Ime fime Window AbsResp SD LRSD Slope RelResp Baseline Keywordsin cyde 1 Value 0g 020 x 02 010 ndng 1 ses 253279 1540 5403 B455 39
159. Biacore X100 Plus Package 186 10 Tools Preferences Users Preferences Users Database Backup User name Full name Group Locked Disabled 8 admin administrator admin No 2 BIACOREIS biacoreis admin No 8 IS Instrument Service admin No 8 NORIO norio user No TRAINING Training uset No Delete Ehange Password Password Properties Password Properties Add User Password never expires Add Biacore X100 Plus Package 10 187 dd User User name Full name Group Description User password Password Retype password C User cannot change password C Password never expires C User must change password at nest login User account C Account is locked C Acco
160. Erdee ycie number x MW adjusted response e Finish Biacore X100 Plus Package 5 139 Evaluation Explorer JEdit x Remove E Sensoreram All sensorerams E Plot gr Baseline Sample B Binding level Binding stability da Binding to reference Carry over Controls binding Controls stakility A Solvent Correction Evaluation Explorer Plot plot Work areq v 100XRU Da g 2 n E Hi 25 Cycle number Biacore X100 Plus Package 140 5 HE 5 5 Evaluation Explorer Plot Baseline Sqmple baseline
161. Keywords in cycle Value 204634 0 05 0 01 00 Yes Baseline Chip CM5 206225 016 006 1591 No ContactTime 420 219185 058 031 14551 No Ligand FlowRate 5 21956 4 0 11 006 14930 No Immobilized Ligand ProteinA Method Amine Procedure TimeAndFlow Temperature 25 00 9C Sensor chip CMS Running standby remaining time 6 9 days Standby Immobilization Results Biacore X100 Plus Package 36 3 V Immobilization Results Chip CM5 Ligand Response Response 1 How cell Procedure Method 1 Bound RU Final RU Time and flow Amine ProteinA i 1296 0 1493 0 Response Bound Response Final RU AE 3 4 Response Bound Final 2 Bound Final NHS EDC
162. 0 0 RU d EE 25 Mines Y a e Reference Line 156 0 s 0 0 RU 8 22500 4 zj Al 21500 4 21000 4 20500 4 Nei 20000 t p t t t ni t 1 0 100 200 300 400 500 600 700 800 900 1000 Time Fc Time Window AbsResp SD LASD Slope HeHesp Baseline Id Keywords in cycle 1 Value 2 1760 5 219 736 007 007 0l 00 Yes baselne1 2 300 5 243976 2183 015 1167 24240 No indng 1 2 360 5 244532 663 013 354 24856 No stabiity 1 He DUEECTY Onine comi Temperature 25 00 c Running manual run amp View Baseline RU 0 Reference Line IE 344 0 s 41 5 RU i 200 400 600 AbsHesp SD LASD Slope ReRiesp Baseine Id 219736 007 00 baseline 1 bindna 1 7 00 00 Yes 243876 2183 015 1167 24240 No ing 1 244582 663 013 354 24856 No stabity_1 Flow 30 Flow path 1 2 Online COMI Temperature 25 00 c Running manual run Mondav October 27 2008 amp
163. 0 1585 3 500E 9 0 4129 7 500E 9 0 1520 1 500E 6 E d 0 5975 3 000E 8 0 1167 Rmax RI Add Fit Parameters LA Parameter Settings Fit Initial value Fit global 1e5 Fit global 1e 3 Fit global YMax Fit global 1e8 Fi local YMaxj5 R R Fit Fit local 0 RI RI Fit 392 AD amp 2 U YIL Constant Initial value 0 Parameter Setting OK Fit
164. 7 Flow 100Cplitmin 10 Flow 10 bunim 11 Flow 100 plimin Time E Included blanks Advanced settings Bac Nest Cancel j Cycle CurVe Ligand Sample Dilution factor Flow ul min Initial rate RU S QC ratio prel Temp C MW Da D m s Blank used 10 5 QC EE Expand all cycles Use reference subtraction data l Biacore X100 Plus Package 156 6 amaro Lehed
165. AB B amp AB2 Heterogeneous Analyte 1 2 A1 B A1B A2 B A2B Heterogeneous Ligand 2 A B1 ABl A B2 AB2 Two state Reaction conformation change A B AB AB Biacore X100 Plus Package 3 77 Fit F5 Kinetics Affinity Fit Kinetics Create AddFit Current Fits Model 1 1 Binding 1 1 1 Bindinc Description il Components Quality Control Report Residuals Parameters Kinetic constants are within instrument specifications Kinetic constants appear to be uniquely determined No significant bulk contributions RI found o Check that sensorgrams have sufficient curvature le Examine the residual plot Pay attention to systematic and non random dev
166. AbsResp SD LASD RelResp Baseline Id 5 36881 6 0 08 010 0 00 00 Ye baseline 1 5 596026 258 023 1 38 227210 binding 1 5 368737 016 014 0 05 31 8 No stability 1 Id baseline_1 10 1 i 1 binding 1 5 PT stability 1 10 t 200 baseline 1 RU o RU RelResp 0 0 binding 1 lt stability 1 RelResp boseline_ 1 72250 RU 2 baseline 2 binding 2 stability 2 RelResp baseline_ 2 RU Biacore X100 Plus Package 2 21 Reference line M View Reference Line ET E ARE ETT lmlal Gu EE aE T Lock sose 5 47082 1 PU sm m E XE
167. By murain B nakayama By Norio Group Bg Shemy Yoshimi E Software exercises E system check administrator Instrument Service Name 2 5 4 1 ProteinA mlgG2 5 9 2007 2 26 05 PM ProteinA mlgG 5 9 2007 9 58 30 AM 070509 5 9 2007 9 27 23 AM Modified 5 8 2007 2 31 33 PM 5 8 2007 2 29 32 PM 5 8 2007 2 08 54 PM 5 8 2007 2 08 04 PM ProteinA m IgG Save Biacore X100 Plus Package 3 27 T admin Biacore X100 Control Software Fle yiew Run Tools Window Help Service3 Assay Worktiw Proteina mlgG X AS Type of assay Direct binding 0 PR Ligand name Proteina ligand Selected chip CM5 Edit Assay Workflow Sensor Surface Preparation Overvi Results reference FindImmobilization pH Bun to find out Enter known values 1 7 Overview Results reference
168. 1 20 48 PM EA Fe Edt View Run Tools Hep DFe F eee E Curve Sensorgram Fo 1 1 1 4 stabiity 2 Finied Start Pos 1 inled Ready 10 ui inled Start Pos 2 inled Ready 10 ui pIniect Start Pos 3 nled Ready 10 ui wash Ready Flowil Start pras Ready 5 wiser zz Online COM1 Temperature 25 00 C Running assay conditions sample V8 Assay Conditions Sample Results RU mouse IgG Fc1 FC2 Fc2 1 Relative Response 9 504 B gg ES E 9 x s 50 t t t t t t 1 0 200 400 600 800 1000 1200 1400 Time s Align curves Fc Name Sample name Conc pg ml RelResp RU 1 binding mouse IgG 0 002 44 1 stability mouse IgG i i 0 6 1 binding 2 mouse IgG 0 02 4 9 1 stability 2 mouse IgG 1 0 4 1 binding_3 mouse IgG 02 51 1 stability 3 mouse IgG i i 0 5 1 binding 4 mouse IgG 2 5 9 Biacore X100 Plus Package 4 117 Results Standby Next gt Assay Condi
169. 28 28 AM 5 9 2007 12 13 59 PM le to the right of the main sensitive support at all E Immobiization pH Scouting 5 9 2007 11 16 37 AM 5 9 2007 11 26 06 AM ks use Blacore X100 Control l mrcikaton pH Scouting 5 9 2007 104729 AM 5 9 2007 110342 4M Trairi I mchizaton pH Scouting 5 3 2007 10 17 39 AM 5 9 2007 10 32 55 AM ini a ProteinA mlgG assay 5 9 2007 10 02 21 AM Training Wizard Template E PlclenMmlgG 5 9 2007 9 58 30 AM Training Assay Woddiow Z aenwnoG regeneration condtion 5 9 200795648AM Training Wizard Template noe sample condition 5 5 200794721AM Training Wizard Template Z aenwnea mmobization 5 9 200793820AM Training Wizard Template ProteinA migG precon 5 9 200793330AM Training Wizard Template EN 070509 5 9 2007 9 2723AM Training Assay Woddiow H i H H 5 8 200723133PM Training Assay Wotdow Training Assay Wotdiow Training Assay Wotdow Training Assay Wordiow 5 8 200720144 PM Taning Wizard Template 5 8 2007 1 5807PM Tanng Wizard Template 5 8 2007 1 47 32 PM Training Wizard Template Training Wizard Template ESIESIESIESJUS mm ERE um 160 E Ue US CNN ie 9 Biacore X100
170. 45 Sliow Curves of Sdme TWD 45 Jar eu Relliac ndeti om 45 SO 148 Solvent correctionh RE RR NR M 136 137 Specify contact time oce I bere t dE e no E deve e eee dede 41 102 Kj S 20 148 149 Standard error oos see RE RACE ARDEN D ERI GOD DAN LOAD DRE e OD e DN cod Dee de Cute 80 Stand bY 6 161 170 Steady StateAffinit ois sisi GE 87 Stop Database ore eR EE EI 171 172 173 Stop RUN A EEE EE 22 30 52 95 113 145 STI Te pof To E rE ereo T a EEE E EEE E VEE TEE aeae o e i TEE a avi E H 166 T temperakufe 1 9 Two state Reaction conformation change lr 76 U Undock mn 6 161 170 171 Usern UL 185 AETERNE ROSSO MM 185 186 187 189 UST EE E E 80 V Vial well position eger EE 16 47 108 W Wizards 102 Work dt6 e ee eer e dee e lt 139 176 177 Workflow e e ec eid e ee x a p e eei ec te se b bae eden 89 0 3 RI DA RAO E N int ettet ductae Mor TTT IP HTS RN giis SM Cte ae B C G 2 C R PIDA FOREN UU NN NE G O LK psy B RR EE D OO H PIETER RR RR E ER B qu Bom m EEA M ANNTIRRRON RTIRITN 19 23 Rn ES EG 12 nU mI uum D EE S C o
171. Baseline Sample Response 5 7 2 Sample Response Baseline 5 2 7 2 Biacore X100 Plus Package 3 61 Gsensorgrams Display Cycles Next gt Close
172. Bun to find out Enter known values 1 7 Overview Results reference Find Sample Conditions Bunto find out Enter known values F 2 Find Regeneration Condiions Bunto find out Enter known values RunkKineticsZAffinty Assay 5 E Temperature 25 00 C Sensor chip CMS Running standby remaining time 7 0 days Run to find out L d Run 2 5 Overview Results reference Enter known values D DREADS DE Overview Biacore X100 Plus Package 4 93 4 2 pH pH iji F
173. Cell 1 Flow Cell 2 d ias ee Regeneration C Use GST Capture Kit Injection Sequence Next gt PE Concentration CFCA System Preparation Prime Prime Prime before run Conditioning Hun conditioning cycle Startup 3 Hun startup cycles Solution buffer Number of cycles Next gt Biacore X100 Plus Package 152 6 Concentration CFCA Injection Parameters x Sample Contact time s First regeneration Solution Glycine HCl pH2 8 Contact time s Stabilization period o s Next gt V Concentration CFCA Samples Sample table Each row in in the sample table will generate two cycles with different flow rates Sample id Dilution factor D 20 C MW Da 1 19E 10 11800 1 19E 10 11800 1 19E 10 11800 1 19E 10 11800 20 C
174. Ethanolamine HCI 1M EDTA lt 0 35 M RESER Surfactant P 20 Tween 20 lt 5 Triton X 100 lt 5 SDS lt 0 5 Octylglucoside lt 40 mM Acetonitrile lt 2096 DMSO 896 Ethyleneglycol in HBS buffer 5096 Ethanol 2096 Formamide 4096 FIR Guanidine HCI 5M Urea 8M Biacore X100 Plus Package 1 IV ii 1 C 1 4 amp l A t jM 7324 ID VS 5 l rd Dl E d Se D vat 5 20 H 200 c eoo
175. Ligand name ProteinA Edit Assay Workflow Sensor Surface Preparation Overview Results reference Fiaa Immobilization buffer 10 mM Acetate pH 5 pahlana phl Scouting All results 3 a Immolslize Method in fc 2 Ami ine Immobilization 5 11 2007 iized in fc 2 451 3 RU GE lmmobil Chip id 070511 0939 All results Ge dms Assay Overview Results reference Sample Conditions Sampl mouse Assay Conditions Sample an Sample Concenra on E ugimi DE 5A172007 12043 PM Contact time 60 s All results t3 A Rntoindon Dissociation time 130 s Regeneration Needed Find Regeneration Conditions 1 Regeneration solution 10mM Gly HCI pH2 0 Contact ime30s Stabilization ime 0 s t3 E Run to find out Enter known values m Run Kinelics Afinity Assay zi aun Help Online coms Temperature 25 009 Running standby remaining time 4 0 days Friday May 11 2007 reg 4 Paint TRAINING amp Biacore fii Kinetics ProteinA ol Lais IGT 3 30PM 2 220 7Z Hk40X Assay Find Regeneration Conditions Run to find out Overview Biacore X100 Plus Package 124 4 4 5
176. OChoose molecular shape Globular 1 2 3 Globular 1 2 Moderately elongated 1 7 amp UV97 V28 Elongated 2 5 O Enter value S Viscosity relative to water at 20 C 20 C O Use standard value 1 00 1 O Enter value lt cnccurnrE Convert diffusion coefficient from temperature T to 20 C Temperature T 25 CC D at temperature T 4 00e 11 mis CONVERT Diffusion coefficient at 20 C 3 49e 11 m s
177. Place 15 ml diluted BlAdisinfectant Solution on the left hand tray and insert the pump inlet 2 BlAdisinfectant Solution 1 5 ml 20 m Start 23 l 3 4 4 Desorb and Sanitize Step 4 Wipe the pump inlet tubes with a moist tissue Place water on the left hand tray and insert the pump inlet tubes Back Close ARARZERBIO 2 7 TERRA VU VFTA 7235 i88CKChNFJU I2 tv FU sS Start 13 Desorb and Sanitize The Desorb and Sanitize procedure is completed Allow the system to run in standby mode for at least 3 4 hours before forming a run 4 Standby stqndby 3 4 Prime 3 Close Biacore X100 Plus Package 166 7 7 2 3 Superclean Supercleqn
178. Plus Package 136 5 HE 5 3 Solvent correction Include F Solvent Correction Chiz RUz YO RU 4iFem21 1 E zoom lock Response Act Ref N E Extrapolation Range Report Point Range T 20 4 1 t 1 None air icu 1400 1200 1000 800 600 400 200 0 200 400 600 Response Ref RUJ LC es Biacore X100 Plus Package 5 137 DMSO DMSO DMSO
179. d e PAETE E EAE E AS E ld hhh 163 7 2 2 Desorb arid Sanitize tte tette ete EER 164 7 2 3 S pefcleata occae oem e Nee lac E 166 13 4 m dtt tmnban es 168 170 8 170 8 1 1 Standby O seesssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssss ns 170 8 12 MEA SOL CRES EEUU LEES 170 82 d 9 T w Jt esca pn eventa b 174 g dq UD AODIEEE cec ottiene 175 9 1 Amo EO E O I o npe eee Oed 175 P spar NONO P P E OS 175 93 p EDESLOBBI ios oso bdo atus 177 9 351 o4 E 92 ILOR D ta i o e tU ee oet odas bnt 177 9 3 2 178 9 3 3 HY TIRMA NLRI L8 euet 178 9 3 4 179 oca KR 180 182 E fus aie 224 EIER ER E E 183 097 184 10 7 8 E e e eed a 185 OS 185 1082 188 Biacore X100 Plus Package Biacore X100 Plus Package A Biacore Biacore surface plasmon resonance SPR
180. eno Time Jd Ko M 1 Ko KA 1 M A B AB Kp A B 48 Biqcore Kp 4 Aa M1s s A B AB Ko Ka Kc Ka Ka Ka Biacore X100 Plus Package J Kop
181. gt Start Biacore X100 Y Y Changes detected in wizard template Help Save Save As Dont Save J Cancel Save As Don t Save Sqve Ctrl Breakl 4 2 95 Standby XYY KOZ Setup Custom Methods T Custom Methods Methods X Aldehyde IX Amine Ligand thiol Surface thiol Method name Biacore X100 Plus Package 104 4 Methods
182. immobilized level Ligand solution Protein in 10 mM Acetate pH 5 Specify contact time Contact time 420 s 420 2 1 2 7 Next gt V8 Immobilization Rack Positions Type 126 Ethanolamine Immobilization 75 Protein Immobilization 85 EDC Mix Fc2 85 NHS Mix Fc 2 Empty EDC NHS min capacity 130H Mix Fc 2 Full Deionized water Water Help Menu Load Samples lt Back ii Next gt Close jil Rack Positions Biacore X100 Plus Package 100 4 Ethanolamine 126 ul 11 mm Ligand 75 ul 11 mm EDC 85 ul 11 mm NHS 85 ul 11 mm Ze NHS EDC 11 mm
183. in wizard template Sqve As RELI Dont Save Sqve 3 2 30 Standby sStqndby 2 1 4 22 Biacore X100 Evaluation Software Biacore X100 Plus Package BE 3 10
184. ley New Cycle 101 24950 24940 e 24930 s H 24920 4 24910 24900 4 0 5 10 15 20 25 30 35 40 45 50 Time Fc Time Window AbsResp SD LRSD Slope RelResp Baseine ld Keywords incyde 1 Value Fow 10 Fon paih 1 Online COM1 Temperature 25 00 C Running manual run Biacore X100 Plus Package 16 2 2 1 1 2 1 17 ineo PS TRAINING Biacore X100 Contrd EA Fie Edit View Commands Run Tc Injection command Az Vial well position Contact time s Minimum required volume in vial well for this injection 50 ul Vial well position amp 2 Vialwellposition 8 Contact time s Minimum required volume in vial well for this injection 50 ul Contact time
185. ul min Multi cycle kinetics Single cycle kinetics Commands in Add Settings for Contact time 120 180 s Dissociation time 120 180 s Flowrate 30 Il min Clycle types solvent correction Biqcore X100 Plus Package 5 131 KE Custom Assay Wizard Cycle Definition Cycle types Binding Analysis Delete Solvent correction Rename Copy Commands in Solvent correction Settings for Solvent correction 1 Available Selected The Solvent correction injection will be run for 30 seconds with a flow rate of 30 Solvent correction 1 Me Sample Solvent correction 2 Enhancement Solvent correction 3 Regeneration Solvent correction 4 Solvent correction Solvent correction 5 Solvent correction 6 Solvent correction 7 Solvent correction 8 Remove lt Back Nest Close Next gt V Custom Assay Wizard Report Points Binding Analysis Solvent correction Sec Befor
186. 0 008 EA Ele gdt View Commands Run Tool Help Service3 Sx xd r0 XE 5 oe E cuve E Sron 3 ER 42 Help m Lock scale 25000 4 1 d ica ME Eu 20000 4 15000 4 10000 4 8 f a 5000 4 04 5000 4 10000 t t t t t t t 1 0 5 10 15 20 25 30 3 40 45 50 Time Fc Time Window AbsHesp SD LRSD Slope HeHesp Baseine ld Keywords in cycle 1 Value Flow 30 Flow path 1 2 Online comt Temperature 25 00 C Running manual run 1 2 2 1 AE 4 6 1 View Show Only Current Curve View Show All Curves View Show Curves of Same Type Biacore X100 Plus Package 4 107 AE 4 7 5 10 30 ul min 72 53 amp 1R
187. 00 ml LR E Bum Wo j i Biacore X100 Plus Package 2 1 1 1 3 BE 1 2 HBS EP 10X 50 ml 4 BR 1008 26 0 1 M HEPES 1 5 M NaCl 30 mM EDTA 0 5 96 v v Surfactant P 20 10 HBS P 10X 50 ml 4 BR 1008 27 0 1 M HEPES 1 5 M NaCl 0 5 96 v v Surfactant P 20 10 HBS N 10X 50 ml 4 BR 1008 28 0 1 M HEPES 1 5 M NaCI
188. 000 105 m IgG Sample 150000 1275 Buffer Startup 105 Buffer Startup 225 10mM Gly HCl pH2 0 Regeneration Full Deionized water Water Toowoon eiu NiE E Menu lt Load Samples lt Back Next gt Close Menu Automatic Positioning Automatic Positioning Change the order in which the samples are positioned by ordering the regions The first region in the list is positioned first Regeneration Nl Brovin Pooling VYes Pooling No OK Biacore X100 Plus Package 3 67 3 4 2 IV ii Run Kineteics Affinity Assay l I amp 2Uv2ULxs PE Kinetics Affinity Injection Sequence Detection Chip Elow cel 12 deference subtraction Chip type CM5 Kinetics type Single cycle Multi cycle Injections in analysis cycle S Ss Flow Celli Flowcellz Sample
189. 20 500 1500 2000 Remove Selection Kinetics gt Biacore X100 Plus Package 76 3 F5 Kinetics Affinity Fit Kinetics Create Add Fit Modet 1 1 Binding Two State Reaction 1 1 Binding Bivalent Analyte Heterogeneous Analyte Heterogeneous Ligand Model 1 1 Binding 3 14 BE 3 14 B A 1 1 Binding A B AB Bivalent Analyte AB B A B AB
190. 3 50 50 50 120 Full Full Full Full Full Full Full Full Sample2 Sample3 Sample4 Sample5 Buffer Solvent correction1 Solvent correction2 Solvent correction3 Solvent correction4 Solvent corrections Solvent correction Solvent correction Solvent correction Full Deionized water Sample 5ample Sample Sample Startup Solvent correction buffer Solvent correction buffer Solvent correction buffer Solvent correction buffer Solvent correction buffer Solvent correction buffer Solvent correction buffer Solvent correction buffer Water jl Rack Positions Next gt Start Sqve Run Stop Run Standby standby 2 1 4 22 Biacore X100 Evaluation Software
191. 3 Assay Workflow ProteinA_m IgG Assay Overview Type of assay analyte Ligand name 9 ligand Selected chip Direct binding ProteinA CM5 Edit Assay Workflow Sensor Surface Preparation Overview FindImmobilization pH brescisizaten bulle 10 iN Acetate cH 5 Immobilize Overview Find Sample Conditions zz Bunto find out Enter known values Find Regeneration Conditions mi Bun to find out Enter known values RunKinetics Affiny Assay Results reference Online COMI Temperature 25 00 C Sensor chip CMS Running standby remaining time 7 0 days Find Immobilization pH B Overview Results reference Find Immobilization pH lt All results Biacore X100 Plus Package 34 3 3 3
192. 35 0 0202969 0 150391 34 0 113281 0 3125 33 32 33 0 104492 0 268555 32 0 154297 0 125977 31 0 106445 0 250586 30 0 0410156 0 291992 29 0 0449219 0 276367 28 0 139648 0 349609 27 0 108398 0 298828 26 0 121094 0 30957 25 24 25 0 146484 0 416992 24 0 355469 0 726563 23 0 455078 0 576102 2 0315324 0 294922 20 21 oo214844 0 023a375 20 0 191406 0 0244141 19 6 83594e 3 0 169922 18 0 60449 0 461914 37 0 283203 0 0546875 16 0 242188 0 380648 15 0 509766 a us j gt Export To other database T admin Biacore X100 Control Software File View Tools Help gre Create Assay Workflow Kinelics Affinit Ea Binding Analysis Other Options LJ Quick Filter Advanced Filter Filtered on Folder test E x Name Modified CreatedBy Type W 8 Users Modified Two Weeks Ago B A administrator E TU amp g lest Kinetios ariri 13 2009 61928 PM 1 9 15 2008 54828 PM adminisslor Evaluation biacorei gt 2 eed Kl Kineics Afiiy s 14 2009 526 10 PM 1 9 15 2009 32718 PM administstor Evaluation E Export To other database Kinetics or Results Search Keys To file I Kinetics Tu lor Results E Kinetics Affinity 9 13 2009 6 19 28 PM 9 15 2009 1 53 23 PM administrator Results Modified February 2009 a
193. 87 ee Te 66 71 Preferences eot lt lt lt lll lt lt lt lt 186 188 Prime 8 54 57 161 168 171 P ERR RR EDEN 23 Q Quality Control 77 Quick Filter e erre ete tete E OS Rae o n e EE M e re Ete e E e RT UR ed 3 4 R Rack Ejected e rr REUNIR ERN EAS ERE EBRE MARRE REMANERE RATEN SENE ES 10 Rack Illumination RE Ei Ei 10 radcklocked E 10 14 Rack Positions rice er Retos Reference line c aree moerore RR iunii de NONU T Ee RUNI 21 Reference Line 21 50 111 Reference subtraction 13 44 105 Regeneration Combmiarid rete t aR 50 111 Miu MicbroaciMe RelResp Report point Dil m Response Bound Response Find R s lt To Excel CE 2 X 9 Results reference ds ReuseChip i Din EE Rmax oooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooo S hum DP 23 Scale ARRIERE ORE 180 uc URDU 80 Biacore X100 Plus Package 192 SensoF el g esa D 5 6 Sensor Chip Maintenance EE SI 160 161 Sensorgram WindoW sssrinin RE 177 178 179 180 182 183 Set TemperatU Fe E OSS NN SIEGE 9 Show AlECUFVeS Eo
194. Concentration 1 Ligand capture Sample Concentration 2 Sample 5 concentrations per cycle Sample Concentration 3 Sample Concentration 4 Sample Concentration 5 Regeneration Regeneration 5 5 Wizard 2 5 Settings for Sample 1 Solution Single cycle kinetics Contact time Dissocial lion time Flow rate Flow path IV i H Regeneration Biacore X100 Plus Package 68 3 V Kinetics Affinity System Preparation Prime Prime before run Prime Conditioning Hum conditioning cycle Startup Run startup cycles Solution Buffer
195. E H L 185 yid day c Pa B MM G 79 88 M N 134 SU HBTE ERU ne sis stet ete eti fedes etie tuti bertus be terti ote tein UE N O 135 137 nU E MU A E OEA 10 SE NN SG SY B C D E 17352 SR TOR ei Eo e ee OEE e AE EE 28 32 93 DJ 33S7 P SIRE E Eo JI ire SECTOR RURSUS RR M HIER EE N E DE a DADE T O E G N 78 UB DM IUE AAAA 35 38 VES BIA ees NEUEM RAE C n TEE S Los p d NORTE 20 es 11 12 44 53 56 91 93 98 99 102 114 117 123 124 141 PLU SB Me 11 Biacore X100 Plus Package XD cOU 3UIc a Hzc Bid 5 8989 22 5 8I18 amp sOSIL CU Sg BURU azt uq uli S 3 N C3 BIEN lise Ik
196. Extrapolate d Fa RR TO 1400 1200 1000 800 600 400 200 0 200 400 600 Response Ref RU Biacore X100 Plus Package 5 135 OK Evaluation Explorer 138 72 AE 5 2 Exclude point Exclude curve Show Sensorgrams Scale Copy Graph Export Curves Exclude point 15 800 600 400 200 0 200 400 600 800 1000 1200 Response Ref Biacore X100
197. GE Healthcare Biacore Biacore X100 Plus Package Ver 2 Instrument Handbook GE imagination at work EA i A Bidcote E 5 ZS A IERE ROLE T M A Is EEE RD B D I i pH EHIRLL ttt F I ABETERBBE oe ep etn Ree bl cem il2 G IV i 48EERERIXE COD IS 60 0D BEER steterit ttt ttssttsn G IV ii NV i amp 45589 7 7 24 F C OTBBTERERIE esent L N i EPEPTRIUIUE RE oet tea e t EO Rd band P 1 1 1 1 NESICUDPOTPOBB cente uu aS 1 1 1 1 E hi EEE Ek 1 1 1 2 Le__1 1 1 13 50 2 9 3 SEI QE W I eus E e eun 2 1 1 4 ee 3 A JATADI SS RS 5 1 2 1 IEE 5 1 2 2 k kk 8 152 3 9 1 2 4 Hc 7JIoiwvbECTOw J DB ee 10 11 2 w 13 IPS 13 E C r AVD rI EAE E AEE E E AAE 16 21 2 OR FS 4I DBI ttt oe o ete tended 20 235 MERO dn eoe hM oL RAS A A UE Ef 22 2 1 4 Standby RS S uaa E IA OM OM t A baa Fl e anas 22 Biacore X100 Plus Package
198. H Blank irr Flow cell 2 immobilize fow cel2 Method Capturing molecule el ligand solution Target level RU Wash solution 50 mM NaoH Blank irr Help Custom Methods Chip type Method be Surface thiol OAim for immobilized level Target level Wash solution 50 mM NaOH Specify contact time OBlank immobilization Next gt Biacore X100 Plus Package 4 103 Rack Position Next
199. M H Biacore X100 Plus Package 193 9 D II A VLC APUD J re ip RAR Ek lt i 2 kA ii LJ K O PRIBEGEERI e oto vetus cen c rn uL A DLL M ML D RA MC D DOE H d 3vdi mdps RR I 79 BERI EEA ki C G I J K 79 87 E h AAPEEE EEE EEEE OE CN MOS E ee Q R S U 152 lt EEE SARITA EASE IEE REST AAEE Y M PE G 179 151 f RR E NU UU MEN NS C P RN B F P 13 34 99 101 C 36 39 101 6 bl i VSZLULk_ ZcAAs Z L UC lbkb S E _ F _ i Z C H bI TM N E N NAON ONTE G H ga G 57 H 50 51 59 62 107 111 119 120 123 kcal NR D EE C Wolve ESSE OS M eo e e e e Eae dete e a e ete 3 l DINAR A EAE E A O AT OAA VEE N A ATE 78 80 0 H 66 71 ESEESE SAE E OE EAIA Nh SI NA RR l 67 72 0 J ERA C II 3 77 Biacore X100 Plu
200. RunBindingAnalysis Assay zi Run Help Online COMI Temperature 25 00 eC Sensor chip CMS Running standby remaining time 7 0 days Find Immobilization pH EJ Overview Results reference Find Immobilization pH All results Biacore X100 Plus Package 4 99 4 3 I i B Immobilize 2Uv2UzsS Immobilization Setup Chiptype CM5 Flow cell 1 E Immabilize fow cell 1 Method Aim for immobilized level Ligand solution 920220 Amine Flow cell 2 Immobiize fowcal2 Method X Amine Q Aim for
201. Sensorgram e B Ba al al al jB ence 3 Report Point Table rd Report Point Table Biacore X100 Plus Package 154 6 AE 6 5 Keyword Table Tools Keyword Table LL Keyword Table 10 Sample 11 Sample 12i Sample 13 Sample 14i Sample 15 Sample 16 Sample 17 Sample 18i Sample l L Concentration Analysis 7 5 1 v 7L i Calbrationfree Biacore X100 Plus Package 6 155 F5 Calibration free concentration Select Samples Create Expand all cycles Show original sensorgrams Sampl Dilution low Initial rate QC ratio Temp MW D mpe factor Cy prel RU s Da mz s ivi iv 9 Fc 2 1 antibeta2micro Sample 500 H 11800 1 362bE 10 1 08 245 1 07 2 16 Fc 2 1 antibeta2micro sample 0 962 25 11800 1 362b 10 5 33 11 3 5 34 View Sample Sample Curve Fc 2 1 mwe Show blank subtracted data RU 120 4 100 4 80 4 B0 4 40 4 Sample Fc 2 1 E Zoom lock 6 Flow 10 ul min
202. Specified in Immobilization Setup EDCNHSINXECT EDC NHS 50 50 Solution Contact Time s Flow Rate l min WASH Ethanolamine LA LIGANDINJECT Specified in Immobiization Setup INJECT Ethanolamine Method Method name Edit V8 Custom Methods Surface thiol Method name Copy of Amine Command Solution Contact Time s Flow Rate ulmin 79 PRECONC Spedfied in Immobiization Setup EDONHSINJECT EDC NHS 50 50 WASH Ethanolamine L LIGANDINJECT Specified in Immobilization Setup INECT Ethanolamine OK Set up Method Biacore X100 Plus Package 44 3 3 4 3 4 1
203. abiy Online COMI ature 25 00 9C Running manual run Injection command 5 Biacore X100 Plus Package 20 2 2 1 2 RU AE 2 3 Fc Time Window
204. arameters M mm v 50 mouse IgG 50 mouse IgG 50 mouse IgG 50 mouse IgG Sample Sample Sample Sample Full Deionized water Water Hep Mens v Lond Samples Sample 1 Conc pg ml Rack Positions Next gt Biacore X100 Plus Package Start l 3 55 Squve 3 2 30 EE TRAINING Biacore X100 Control Software Assay Conditions Sample 5 11 2007 1 20 48 PM EA fe Edt View Run Tools Heb IE ee nled Start Pos1 inied Ready 10 pi Pinied Start Pos 2 pinied Ready 10 pi ined Start Pos 3 biniec Ready 10 pi Flow Start B Wear Ready Online COM1 Temperature 25 00 eC Running assay conditions sample V8 Assay Conditions Sample Results Relative R
205. aseline i Binding level ia Binding stability a Binding to reference E Bar Chart g Report Point Table Report Point Table Bar Chart a7 Conc ug ml Response Relative Response m Bar Chart amp Ctrl Evaluation Explorer ix Remove Edit p Sensorgram a All sensorgrams 5 Plot p Baseline ia Binding level Sample a Binding stability Sample B Binding to reference Sample g Report Point Table ui Binding Analysis Biacore X100 Plus Package 128 5 5
206. bration points Calibration Curve Calibration Curve Analyte name Calibration points Concentration 6 Next gt Biacore X100 Plus Package 144 6 V8 Concentration Analysis Samples Sample table tito factor sample1 1000 sample2 500 sample3 200 sample4 50 sample2 500 sample3 200 sample4 50 Samples Sample table Sample id Dilution factor sg
207. click OK OK BIO Injection command A amp Z2 Vialwell position I 8 Lo Contactime 60 cres Minimum required volume in vial well for this injection 50 ul Biacore X100 Plus Package 110 4 EA Ele Edt View Commands Run Tools Help Servke3 a x xd 79 c E 5 Cycle 1 Cuve Subtacted Fo 2 1 DEN RU E Lock scale 4000 x d z i 4 New Cycle 2021 E 9h Iniect 11120 3500 3000 E H 2500 a 2000 1500 g i 1000 t Ai k t 1 o EI 100 180 200 250 300 350 400 450 s Fc Time Window AbsResp SD LRSD Slope ReResp Baseine Id Keywordsincycle 1 Value 21 1760 5 12945 005 006 000 00 Yes baseine1 21 3010 5 37187 2176 015 11 63 24242 No binding 1 21 3160 5 3755 649 018 347 24810 No stebilly 1 Close UEECTUNE mcm Online COM1 Temperature 25 00 ec Running manual run Fc 2 1
208. d solutions from Maintenance Kit BlAdesorb solution 1 BlAdesorb solution 2 about 10 ml BlAdisinfectant solution about 15 ml Next gt 1 Desorb and Sanitize Step 1 Place 10 ml BlAdesorb Solution 1 on the left hand tray and insert the pump inlet tubes 2 BIAdesorb Solution 1 1 10 ml Start 8 Biacore X100 Plus Package 7 165 1 2 2 Desorb and Sanitize Step 2 Wipe the pump inlet tubes with a moist tissue Place 10 ml BlAdesorb Solution 2 on the left hand tray and insert the pump inlet tubes 2 BIAdesorb Solution 2 2 10 ml Start 7 2 3 3 Desorb and Sanitize Step 3 Wipe the pump inlet tubes with a moist tissue
209. e Single cycle kinetics 2 analytes File View Evaluation Tools Window Help P A cj Sensemram LA Plot p Bar Chart 3 Kinetics Affinity LA Concentration Analysis 1 E All sensorgrams BEIE Edit ET PE KI Curve Name Fc 2 1 me Cycle Purpose Overlay me Cycle Overlay e ensorgram All sensorgrams RU Sensorgram E Plot 450 B Baseline E Report Point Table rjj Report Point Table E Zoom Lock 1000 1500 2500 3000 3500 Thursday October 30 2008 Biacore X100 Evaluation Software A Kinetics Affinity 3 11 73 Biacore X100 Plus Package fip 3 11 Keyword Table Tools Keyword Table Fs Keyword Table Cycle Cycle Purpose Conc pg ml MW Da v v Y v 1g Sample 3 73 Biacore X100 Plus Package
210. e After Start of End of Inject Window Baseline baseline 10 Before Start of Sample 1 10 Before End of Sample 1 20 After End of Sample 1 X Next gt Biacore X100 Plus Package 132 5 KE Custom Assay Wizard Sample Table Binding Analysis Solvent correction Solvent correction Buffer Buffer Buffer buffer Sample 1 Sample2 Sample3 Sample4 f 0000000000 OO OO 0 0 0 0 OOOOOOQ9OOO oooooooooo OO OO O O O O O O undefined O Sample5 Sample 5 Next gt Biacore X100 Plus Package 5 V Custom Assay Wizard Rack Positions 13
211. erature 25 00 C Running manual run IMondav October 27 2008 Biacore X100 Plus Package 4 113 New Cycle iz Commands New Cycle 2 Y w New Cycle Flow am Flow rate ulmin Flow path O E Flow path 1 O IE Flow path2 Reference subtraction Flowpat 12 Flow path OK End manual run 3 Run gt Stop Run Biacore X100 2 This will end the manual run when the last queued command is completed OK Standby Biacore X100 Plus Package 114 4 4 4 2
212. esponse p mouse IgG Fc1 FC2 Fc2 1 v 9 504 EEJ P3 u a 3 0 r 50 t t t t t 1 0 200 400 600 800 1000 1200 1400 Time s v Align curves Fc Name Sample name Conc pg ml RelResp RU 1 binding mouse IgG 0 002 44 1 stability mouse IgG i 0 6 1 binding 2 mouse IgG 0 02 49 1 stability_2 mouse IgG i 0 4 1 binding 3 mouse IgG 02 51 1 stability 3 mouse IgG i 0 5 1 binding 4 mouse IgG 2 5 9 Back Next gt Biacore X100 Plus Package 56 3 Results Standby Next gt V Assay Conditions Sample Save Settings Use these settings in the workflow steps Sample Name mouse lgG Contact time 60 s Dissociation time 130 8 Max concentration 20 v v Regeneration is needed Help Save Settings Save FS TRAINING Biac
213. etics Affinity Fit Kinetics Create AddFit Model Heterogeneous Ligand 1 11 EA N m Parameters il Delete Components Report Residuals Parameters Curve kal 1 Ms kd1 1 s KD1 M ka2 1 Ms kd2 1 s KD2 M Rmax1 RU Rmax2 RU Conc M tc 8 868E 5 0 001174 1 324E 9 1 023E 6 0 003565 3 485E 9 17 09 23 56 1 766E 9 1 000E 9 3 500E 9 7 500E 9 1 500E 8 3 000E 8 lt Back Einish Cancel J Current Fits Current Fits Finish Biacore X100 Plus Package 3 83 F5 Kinetics Affinity Fit Kinetics Create Current Fits 1 1 1 Binding 500 0 500 1000 1500 2000 2500 Time Quality Control Report Residuals Parameters Curve ka 1 Ms kd 1 s KD M Rmax RU Conc M tc Flow ul min kt RU Ms RI RU 1 576E 6 0 003258 2 067E 9 40 00 1 641E 7 d 1 000E 9 30 00 5 100E amp 7
214. ftware Kinetics Affinity 5 14 2007 2 08 23 PM Support Navigator lt lt Print What do you want to do Evaluate kinetics affinity Evaluate binding analysis Evaluate concentration analysis Cycle Pupose Overlay w PH 6 Cycle lt Overiay gt Sensorgram Sensorgram window This window is displayed automatically when you open data for evaluation Click the Sensorgram button to create an additional sensorgram window Use the sensorgram window to examine the sensorgrams for interaction characteristics and data quality Exclude any sensorgrams that are clearly disturbed or deviate from the expected behavior Cycles that you exclude at this stage will not appear in subsequent evaluations Using the sensorgram window Screen regions and menus Using the Support Navigator Biacore X100 Plus Package BE 3 9 IE Kinetics Affinity Rack Positions 915 m IgG Sample 105 m IgG Sample 150000 105 m IgG Sample 150000 105 m IgG Sample 150000 105 m IgG Sample 150
215. g stability ia Binding to reference E Report Point Table Report Point Table Zoom Lock A Kinetics Affinity 3 11 73 FS Kinetics Affinity Select Curves Create Curves Sample LNFP 1 v Temperature 25 Curve Fc 24 me Cycles Sample aM rit 4 LNFP 1 0 30 5 LNFP 1 500 30 6 LNFP 1 250 30 7 LNFP 1 30 8 LNFP 1 30 9 LNFP 1 30 10 LNFP 1 30 11 LNFP 1 12 LNFP 1 n tact Time Diss Time S 60 0 KI KI K KI KI KI KI Kj Zoom lock 60 40 20 Show concentration series Show blank s Show average blank s Multiple Rmax Adjust Injection Start Biacore X100 Plus Package 86 3 3 12 74 lt Next gt F5 Kinetics Affinity Select Data Create Curves Conc Flow Contact Time Diss Time em eer ais uM ml min 6
216. gt Biacore X100 Plus Package 3 75 FS Kinetics Affinity Select Data Create Curves Flow Contact Time Diss Time Edit Cycle Curve ul min s s 30 1 80 0 1800 7 Blank Subtracted Sensorgrams 1500 Time csa 0 0 3 13 AE 3 13 1 Blank Subtracted Sensorgrams
217. he calculation of diffusion coefficients in Calibration Free Concentration Analysis assays Only for users of Biacore X100 Plus Package 2 0 or later valid product key required Access the on line tool here Diffusion Coefficient Calculator Tool here Diffusion Coefficient Calculator Converter This on line tool is designed to help you calculate diffusion coefficients for use in Calibration Free Concentration Analysis assays Itis accessible only via a valid product key associated with the appropriate types of Biacore software Calculate diffusion coefficient at 20 C Molecular weight 90000 Da Frictional ratio Q Choose molecular shape gt Globular 2 T Q Enter value 2 Viscosity relative 9 Use standard value 1 00 to water at 20 C c O Enter value 4 00 caLcuLare Diffusion coefficient at 20 C 6 03e 11 M m s Convert diffusion coefficient from temperature T to 20 C Temperature T 25 CC D at temperature T 4 00e 11 i ms convert Diffusion coefficient at 20 C 3 49e 11 m s Biacore X100 Plus Package U 20C Molecular weight Ddl Friction ratio FRK
218. i noo 1200 1 acetic acid i 1200 0 2 M sodium bicarbonate Reagent 600 6 M guanidine HCl or 50 DMSO Reagent 600 10 mM HCI or 10 DMSO Reagent 1 Full Deionized water Reagent 1 Help Menu v Load Samples lt Back Start Close 1 1 200 uL 0 2 M 1 200 uL 6 M 5096 DMSO 600 uL 10 mM 10 DMSO 600 uL Full Stat Superclean Standby 3 4 RARES DH Prime 3 OF Biacore X100 Plus Package 168 7 7 3 Desorb and Sanitize 35 cd
219. iacore condiyion sample 19 TRAINING Biacore Kinetics_ProteinA 1 Biacore X100 Plus Package 60 3 Regeneration Scouting Results Sensorgrams A Sample Response Baseline Result Standby Trend Chart 9 7 Sample Response Baseline Display 1st cycle 1 Conditions Sample Response Baseline RU 1 Sample Response Baseline 2 B25 4 1
220. iations lt Back Finish Cancel Ji 1 1 Binding Quality Control WE 3 15 Quality Control 5 Q Q Quality Control Report Residuals Parameters O Kinetic constants are within instrument specifications Kinetic constants appear to be uniquely determined eo No significant bulk contributions RI found o Check that sensorgrams have sufficient curvature 6006 0 8 O eo Examine the residual plot Pay attention to systematic and non random deviations Biacore X100 Plus Package 78 3
221. ind Sample Conditions gt Run to find out F T Enter known values Assay Conditions Sample Injection Sequence 1 2 Reference subtraction UE IP I Ee Flow Cell 1 Flow Cell 2 abd Ei AT 5 2 Find Regeneration Step Next gt Biacore X100 Plus Package 54 3 Assay Conditions Sample System Preparation Prime Prime before run Conditioning Run conditioning cycle Prime before run Prime Next gt V8 Assay Conditions Sample Injection P
222. ine X cal i is0 O0 U TET 15606 00 UJ E g FE HE y Ba M e x Reference Line X 156 0 s 0 0 RU 8 E ooo Jf 22500 J 7 22000 4 x m 21500 T 21000 4 20500 4 Nel 20000 t 1 t ni t 5 10 20 500 40 so 609 7m 909 100 Time Fc Time Window AbsResp SD LRSD Slope RelResp Baseline Id Keywords in cycle 1 Value 2 1760 5 21972285 007 007 00 00 Yes baseline 1 2 301 0 5 2439756 2183 015 1167 24240 No inding 1 2 3160 5 244582 6 63 013 354 24856 No stability 1 Hel TERTIA Temperature 25 00 oc Running manual run Online CoM1 View Baseline RU 0 T admin Biacore X100 Control Software Manual Run 10 27 2008 2 14 43 PM EA Fe Edt View Commands Run Tools Help Service3 Dyde Cuve Sensorgram Fc 2 xw Tert ise 2 Hep Reference Line X 344 0 s 41 5 RU Reference Li 344 0 s 41 5 RU E 200 400 800 AbsResp SD 219736 007 243976 21 83 24532 663 LRSD ReResp Baseline Id 24240 No 24856 No stabiity_1 Keywords in cycle 1 Value Flow 30 Flow path 1 2 Online COMI Temp
223. is E 0 DOM Manual Run How mm Flow rate 10 j umin How path Flow path 1 O Flow path 2 Reference E3 m subtraction O E Flow path 12 21 Manual Run Flow rate 5 10 30 ul min 10 min 30 ul min Flow path Flow path1 1 Flow path2 2 Flow path1 2 1 2 Reference subtraction 2 1 Biacore X100 Plus Package 14 2 Load Samples Rack Unlocked Rack Unlocked Insert the rack and click OK rack locked OK Man
224. l SE SE E Avg 23 sample2 27 sample2 0 03213 Avg 24 sample3 28 sample3 Avg 25 sample4 29 sample4 0 01305 Relative Response Avg 1 Sample Table Cycles Sample id Dil Fact ResponselRU Calc Conc CV Calib Curve Finish Biacore X100 Plus Package N A Calc Conc Evaluation Explorer n x Remove v Edit In Sensorgram p Baseline a Binding stability p Report Point Table rjj Report Point Table 6 151 6 2 Other options 1 wizards Concentration Calibration Free CFCAA V Concentration CFCA Injection Sequence Detection Chip Flow cell M Reference subtraction Chip type Injections in analysis cycle Flow
225. lized level RU 34 36 Immobilize Run Immobilization Setup Chiptype Ci 7 Pme before run Prime How cell 1 immobilize fiov cell 1 Method Aim for immobilized level Ligand solution Amine gt rcontact time Flow cell 2 z immobitize fow cel 2 Method E Amine v 1 2 7 Next gt Immobilization Rack Positions 70 50 mM NaOH Immobilization 126 Ethanolamine Immobilization 85 EDC MixFc2 85 NHS Mix Fc 2 Empty EDC NHS min capacity 130ul Mix Fc 2 Full Deionized water Water Rack Positions
226. luation Name Save Biacore X100 Plus Package 10 185 10 10 1 Biacore X100 Control Software Administrators Administrators Users 1 Administrator Username admin Password administrator Administrators Users Administrators
227. ly HCI pH2 0 Contact ime30s Stabilization ime 0 s t3 E Run to find out Enter known values m Run Kinelics Afinity Assay zi aun Help Online coM1 Friday May 11 2007 3 30PM Assay Find Regeneration Conditions Run to find out Overview Biacore X100 Plus Package 3 63 3 4 1 2 Run Kinetics Affinity Assay Run Chp n Chip type CM5 Single cycle Mulicyele Injections in analysis cycle Flow Cell 1 Flow Cell 2 Injection Sequence 9 Kinetics type C Multi cycle Next gt V4 Kinetics Affinity System Preparation Prime Prime before run Prime Conditioning Run conditio Startup v Run startup cycles so Next gt Number of cycles Biacore X100 Plus Package
228. n Software Concentration Analysis 6 2 BE 6 2 727 24 FI SSODEREB Keyword Table Tools Keyword Table fF Keyword Table 18i Sample Biacore X100 Plus Package 148 6 _ o E Concentration Analysis Create Calibration Samples Calibration Curve Settings How Cel Fc 2 Report Point stability 2 4 Parameter v Calibration Table Curve Cycle Conc pg m 4 i Response Calc Conc CV Ru pg ml 9e 0 i DH 21 N A 20 N A MA 118 5 03182 118 2 0 3173 118 3 0 3178 0 1884 236 4 0 6428 235 7 0 6408 236 0 0 6418 0 2220 Relative Response 462 6 1 263 462 0 1 261 462 3 1 262 0 08591 Parameters for curve 1 Rhi 1 11E 04 Rio 8547 A1 253 A2 1 052 Chi 75 48 Calibration Curve Settings Flow cell
229. oasis 149 Evaluation Explorer eter isi 79 88 127 150 176 184 Excl de point a e IEEE NR EE 135 Export s Te 611 137 F OoWipath p p 13 44 52 105 113 FIOWirate iin EE 13 G icr nm cs 181 H Hete rogerieous Analyte RS 76 Heterogeneous Ligand P 76 I Immobilization pH Scouting le F 32 97 Immobilization Results c rre rote tret beata i bna Lettre eR PINE beet XU TR e eee I edes 35 38 Immobilize 3 5 cette terret EE kh tiem Ei 34 37 39 91 99 101 Injection comirridrd rrr teet hss 16 18 19 47 48 108 109 Biacore X100 Plus Package Keyword Table tee nee EI 73 124 147 154 PATATE NNUS 75 88 Kinetics Affinity L Legend eee e rm HER DARE E ERREUR ED et EQ eset N An rat 181 Eodd Sarnples tetto oeei URINE MICE SRI E E aaas 10 14 18 48 109 M Manual RUN EE RR EH EAR EA DRE ISDN ERAT UR CERO EO DO ORE RESTE AUS CRT eu 13 14 44 105 N NES niens ERREUR ERU SON fe nn UE NR TOSS ERRARE OI B D E 32 35 38 100 O Oracle Datab se s interit hai ie te hi tete tete te e bed 171 172 173 188 Other Opti nS coercet tin kk 41 44 105 128 141 151 OVEVI W BRANO 27 33 36 39 56 62 92 98 101 117 125 141 P PadssSWoFd ERREUR 185 186 1
230. ore Word Pad Paint Word Pad 6 REO RAO 0 A470 Dag aj selel Ms PI apo BS J s z v e amp a Ea a e T deemed Fru Adjusted ensorgram 3 AL 200 E B E m s ka 150 s E I E y a J 5 E F1 gt 72 Biacore X100 Plus Package 9 183 9 6 Sensorgram window Export Curves txt Microsoft Excel
231. ore X100 Control Software Assay Workflow ProteinA m IgG Ligand Attachment Overview Type of assay Direct binding ans Selected chip cms ligand name ProteinA ugana Sensor Surface Preparation Overview Results reference i llization pH ng FRR Immobilization buffer 10 mM Acetate pH 5 Be Enotilestion pH Sau ud Run to ind out Method in fc 2 Amine Immobilization 5 11 2007 Immobilized in fc 2 451 3 RU NIAM Chip id 070511 0939 Assay Conditions Sample Find Sample Conditions 5 11 2007 1 20 48 PM E Run to ind out Enter known values Find Regeneration Conditions Run to ind out Run Kinetics Afinity Assay Temperstue 25009 Running standby remaining time 4 0 days Assoy Find Sample Conditions Run to find out Overview Biacore X100 Plus Package 3 57 Find Regeneration Conditions Run to find out Enter known values
232. s Package AR 78 LEDA ian TA ODYS NN NYS NANAvSSNYYAA l b kC h O 161 P B 29 30 70 P UE VN A EE REUNIR NEED 3 a 2 DIL MN CM 32 ESN C P 141 Bc M 10 ASSAI M N TE C G J Q ee 76 82 ASS FG EADE rbd Ra E E E TA A A TA B S EE Cu PP AA NORA AEEA ISE EEI SER EIRO ESE OAE LEOA EEEE EN Cj SIE SR AED E EEAO RE MED ALT CN LK ED NM FU MEX CIAR DX A BE HERR HM J 140 IMMER rei eR A URS RN NE ERN RN SERENA RE IRAE H TB TE EI ee etse eie TR RR ER ERU E RR DER EH eR UE RERUM N ew pm 168 160 Epo R Biacore X100 Plus Package 195 Q R C Q R 156 ro dani IET PS cid SRI RECENTE 11 KK 44 72 3 OO
233. s Window Help gt gt Print Sensooam Plot Kinetics Affinity ai Binding Analysis L Concentration Analysis What do you want to do Evaluate kinetics affinity Cycle Purpose Ovetay gt v Evaluate binding analysis Sensorgram Evaluate concentration analysis Dm sensorgrams x Remove A Edit Sensorgram window This window is displayed automatically when you open data for evaluation Click the ram button to create an additional sensorgram window Use the sensorgram window to examine the sensorgrams for interaction characteristics and data quality Exclude any sensorgrams that are clearly disturbed or deviate from the expected behavior Cycles that you exclude at this stage will not appear in subsequent evaluations Using the sensorgram window Screen regions and menus Using the Support Navigator V8 TRAINING Biac TRAINING Biac Biacore X100 Plus Package 4 jBarChat admin Biacore X100 Evaluation Software Multi cycle kinetics Bar Chart Fle View Evaluation Tools Window Help S e A Sensomram Plot ai Bar Chat A Kinetics Affinity LA Concentration Analysis E Sensorgram A All sensorgrams fg Plot B
234. se V amp S TRAINING Biacore X100 Contrd EA Fie Edit View Commands Run Tc Injection command Vialwell position I 8 ok Contact time s ie Minimum required volume in vial well for this injection 50 ul Vial well positionxD f amp 2 U vy 2U 74 24 F amp t v LEHEL v OA EEU Inject Vial position 11 6 Minimum required volume in vial for this injection 100 ul Contact time 60 120 W IV i G OK Biacore X100 Plus Package 4 109 Cqncel Inject Load Samples P1 IYL Rack Unlocked Rack Unlocked Insert the rack and
235. sition 1 position Position 15 HzO 1 Position H20 Rack position 1 15 Rubber caps type 2 BR 1004 11 1 5 ml Plastic Vials 15 mm Plastic Vials BR 1002 87 BR 1006 54 Biacore X100 Plus Package 1 11 1 3 Biacore X100 3 TRAINING Biacore X100 Control Software Create Assay Workflow Other Options Kinetics Affinity Binding Analysis IZ Wizards Manual Run 2
236. tions Sample Save Settings Use these settings in the workflow steps Sample Name mouse lgG Contact time 60 s Dissociation time 130 8 Max concentration 20 v v Regeneration is needed Help Save Settings Save T TRAINING Biacore X100 Control Software Fie View Tools Help WO F Assay Workflow ProteinA m IgG Ligand Attachment Overview Type of assay Direct binding 0 ame Selected chip CM5 Ligand name ProteinA hana Sensor Surface Preparation Overview Results reference Immobil buffer Acetate Immobilization pH Scouting Find Immobilization pH AR 5512007 101437AM All results gd Run to find out Method in fc 2 Amine Immobilization 5 11 2007 Immobilized in fc 2 451 3 RU DS 150 134M Chip id 070511 0939 Find Sample Conditions Run to find ou Enter known values Dissociation time 130 s Find Regeneration Conditions Run to find out Enter known values Run Kinetics Afinity Assay re Help Online COM1 Temperature 25 00 oC Sensor chip CMS Running standb
237. ual Run Start Save Results As Save in folder Folder Users Name B Assay Conditions Sample 4 3 2007 1 33 49 PM 4 24 2007 7 15 administrator Kinetics Affinity 4 3 2007 2 51 26 PM 4 24 2007 7 15 Instrument Service Il Singe cycle kinetics 2 analytes 4 24 2007 7164 Assay Conditions Sample 1 4 24 2007 7 16 Binding Analysis b2u 4 24 2007 7164 E Binding Analysis Sandwich Assay 4 24 2007 7 161 Concentration Analysis 4 24 2007 7 164 Immobilization Conc 4 24 2007 7 161 E Immobilization pH Scouting 5 9 2007 10 17 39 AM 5 9 2007 10 324 E immobilization pH Scouting 5 9 2007 10 47 29 AM 5 9 2007 11 03 Immobilization pH Scouting 5 9 2007 11 16 37 AM 5 9 2007 11 261 E Immobilization 5 9 2007 11 28 28 AM 5 9 2007 12 13 IE Immobilization 4 25 2007 11 04 44 AM 4 25 2007 11 22 t Manual Run 5 10 2007 1 30 44 PM save Caneel Save Biacore X100 Plus Package 2 15 EA Fe Edit View Commands Run Tools Help EHE D FPO FE code Cuve Sensorgram Fe s 8 i t boe Lock scale 24960 4 ic Y
238. ues a eren Jem Immobilize Find Immobilization pH Run to find out Enter known values V Immobilization pH Scouting Setup pH 10 mM Acetate 10 mM Acetate 10 mM Acetate Next gt Biacore X100 Plus Package 3 29 PE Immobilization pH Scouting Injection Parameters Prime Prime before run Prime Liqand Name E 60 Surface regeneration This surface wash will be run once at the end of each cycle 50 mM NaOH Next gt V8 Immobilization pH Scouting Rack Positions 45 ProteinA Sample 10 mM Acetate 5 45 ProteinA Sample 10 mM Acetate 4 5 45 ProteinA Sample 110 mM Acetate 4 135 50mM NaOH Regeneration Full Deionized water Water Rack Positions
239. uments amp Pte ae X Biacore X100 Control ye Favorites Software 1d My computer Biacore X100 Evaluation Software B Lacs m e a 9 Help and Support Search HP support B Log Off Turn Off Computer pos Oracle Database The OracleServiceXE service was stopped successfully DOS amp Stop Database REM BN E oppin s SS LS as stoppe C Norac lexe app orac TeNproductN1 2 8NserverNBIN gt Windows Shutdown Biacore X100 PC Oracle Database Biacore X100 Plus Package 8
240. unt is disabled User name Full name Group Administrators Users Description User password Password User cannot change password Password never expires User must change password at next login 1 9 528IBDO2Z 4UfisEIE VA72 FOZSB aqU dg 007 User account LlAccount is locked Account is disabled OK Biacore X100 Plus Package 188 10 10 2 Biacore X100 Control V Biacore X100 Evaluation
241. y remaining time 4 0 days E EA TRAINING Biacore condiyionsample9 TRAINING Biacore Assay Find Sample Conditions Run to find out EJ Overview Biacore X100 Plus Package 118 4 Find Regeneration Conditions Run to find out Enter known values V8 Regeneration Scouting Injection Sequence Detection Flow cell Injections in analysis cycle FowCelli Fowcemz 2 Next gt V8 Assay Conditions Sample System Preparation Prime Prime before run Conditioning Run conditioning cycle Prime before run Prime Next gt Injection Parameters Sample Find Sample Conditions
242. z E x B i PEE p ji eg T5 g E Bii i S Bir i 99 E H Sei i EH g HH zai 00 00 1500 2500 Sensor chip CMS Running standby remaining time 4 0 days Aim for immobilized level Standby Standby 2 1 4 22 Immobilization Results Biqcore X100 Plus Package Immobilization Results Flow cell Procedure Method 3 39 2 Targetlevel Amine Yes ProteinA Response RU Final RU Target Reached 451 3 i Response Bound Response Final RU 3 4 36 Immobilization Result Close Close jw I admin Biacore X100 Control Software File View Run Tools Window Help Service3 SGuY X Ee 28 Assay Workflow ProteinA_mlgG Assay Overview Type of assay

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