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Biacore X100 Plus Package 取扱説明書

Contents

1. Req vsC CO Req KA KD Req C x Rmax C Ko Req RU 4 Conc M Biqcore X100 XI 5 0
2. RI 0 RU Residual Residuals Y Residuals for a good fit Residuals for a poor fit RU RU Response 100 100 z200 300 400 s00 100 100 200 300 400 Time amp Time Z Chi U value ITENS C SNEDE E 15 25 SE Standard error Parameters SE SE
3. Sensor Chip Maintenance 25C BIA maintenance Kit BR 1006 66 BIAdesorb solution 1 90 ml BlAdesorb solution 2 90 ml BlAtest solution 65 ml BlAdisinfectant solution conc 10 ml BlAnormalizing solution 30 ml Sensor Chip Maintenance 1 BIAdesorb solution1 4C Biqcore X100 6 137 6 1 Tools gt Undock Chip Cl l Biacore X100 This will undock the sensor chip Help Undock Chip Cancel Undock Chip Sensor chip
4. Restore OK Cc Biacore X100 leg bud E ads ET din i ffIIHIEHII Users 1
5. 5 200 ug ml 10 mM Immobilization pH Scouting 50 mM NaOH BiacoreeX100 VII D D 1 THE TERBAWE OSEE S
6. Blank Subtracted Sensorgrams Remove Selection Kinetics gt BiacoreeX100 3 75 Kinetics Affinity Fit Kinetics Create Add Fit Model 1 1 Binding v Bivalent Analyte Heterogeneous Analyte Heterogeneous Ligand Two State Reaction ime s Model v ETOR 1 1 Binding 3 13 3 13 B A 1 1 Binding A B AB Bivalent Analyte AB
7. 3 4 1 Otheroptions b Manual Run 9 I v2d 2 Manual Eun Flow F Flow rate EI v imin Flom path Q Flow path 1 Q fed Flow path 2 Reference subtraction 5 Flow path 1 2 30ul min Flow path 1 2 Reference subtraction 2 1 Start Sqve BiacoreeX100 3 45 admin Biacore X100 Control Software Manual Run 10 27 2008 2 14 43 PM i 12 jep Li 10000 5 10 15 Fel Time Window AbsResp SD LRSD Slope RelResp Baseline ld E Keywords in cycle 1 Value l Flow 30 Flow path 1 2 Online COM1 Temperature 25 00 9C 1 2 2 1 3 7 1
8. 30 1 112 URBS 7 b62 U 78380 o INDUBS ESI 1 DAA CSI S S pH m NaCl lt 2M REESE 10 mM Gly HCI pH 1 5 HCI 100 mM Phosphoric acid 100 mM Formic acid 20 96 PILH Y RI 10mM Gly NaOH lt pH 12 NaOH 100 mM Ethanolamine 100 mM Ethanolamine HCI 1M EDTA lt 0 35 M Surfactant P 20 Tween 20 5 96 Triton X 100 lt 5 SDS 0 5 96 Octylglucoside 40 mM Acetonitrile lt 20 DMSO lt 8 Ethyleneglycol in HBS buffer lt 50 Ethanol lt 20 Formamide lt 40 Zr RI Guanidine HCI lt 5M Urea 8M BiacoreeX100
9. Final Bound NHS Final Bound Immobilization Result Close Close t g Hiaconr X108 Cinbo Be Wee un Tech yei Lenses D I T T Immobilize 69 Overview Z Results reference Biqcore X100 5 111 5 5
10. amp lt Y Biacore X100 3 47 23 24 FORD 2 1 3 Hep TRAINING Biacore X100 Contr I File Edit View Commands Run Tc m 43 EB Cycle 1 2 Heb Vialwell position Il Contact time s Vial well position Inject Val position 11 a Minimum required volume in vial For thie injection 100 pd Contact time 60 120 M D 1 VII OK Biqcore X100
11. RI Residuals Y Residuals for a poor fit Residuals for a good fit Response 100 100 200 300 400 sgg 100 1 z200 300 400 Time amp Time Z Biqcore X100 78 3 Report 97 Hualitu Control Report Residuals Parameters EE a 1 576E 6 0 003258 2 067
12. AJ Kinetics Affinity Biacore X100 All rz 4 4 1 Concentration Analysis Binding Analysis Kinetics Affinity Overview D 3 Xll Concentration Analysis Injection Sequence Detection Chip Elow cel Chp ape Injections in analysis cycle Flow Cell 1 Flow Cell 2 Cen sme Enhancement Regeneration Injection Sequence Detection Flow cell Chip Chip type Injections in analysis cycle Capture
13. crude Bulk Effect 5S ONG RB ng ml
14. Quick Filter Advanced Filters E Biqcore X100 4 1 1 3 Quick Filter xj 3 Biqcore X
15. Final Bound Immobilization Result Close Close Hacor X100 Enmhaml Be Wee un Tech yei Lenses D I T T Immobilize 69 Overview Z Results reference Biqcore X100 3 37 Immobilize Run Immobilization Setup x Chiptype Prime before run Prime How cell 1 lmmcpii Method Ligand solution Do Amine ow cdl2 Immobitize flow cell 2 Method
16. Users Administrators Biacore X100 Control Software Administrators Tools Preferences Users BiacoreeX100 160 9 Preferences Users Database Backup User name Full name Group Locked Disabled 2 admin administrator admin No 2 BIACOREIS biacoreis admin No a IS Instrument Service admin No 2 NORIO norio user No 8 TRAINING Training user No Password Properties Password Properties 2 U v 2d S
17. Next gt Concentration Analysis Rack Positions 125 m IgG Calibration 125 m IgG Calibration 125 m IgG Calibration 125 m IgG Calibration 125 m IgG Calibration 125 m IgG Calibration 125 m IgG Calibration 70 sample1 Sample 70 sample2 Sample 70 sample3 Sample 290 buffer Startup 855 10mM Gly HCl Regeneration Full Deionized water Water gt Rack Positions Next gt Biqcore X100 92 4 Start Biacore X 100 A Changes detected in wizard template Save As Dont 5ave Cancel Save As Don t Save Ssave 2U v 23g S GRE SERISIEUIEUSSI amp 4 1 E 4 1
18. Include A Conc Flow mers Time S 1 Next gt Biqcore X100 74 3 F3 Kinetics Affinity Select Data Create Curves Edit cycle qup PERS Time UR eI Blank Subtracted Sensorgrams E Zoom lock 0 0 3 12 3 12
19. Biacore X100 112 5 Rack Position Next gt Start Biacore X100 A Changes detected in wizard template Save As Don t Save Sqve 5 2 104 Standby O Biacore X100 5 113 Methods
20. OK Hidden OK Gridlines Gridlines m AMS Major Gridlines Minor Gridlines Y AMS Major Gridlines Minor Gridlines Major Gridlines Minor Gridlines OK BiacoreeX100 156 8 8 5 Sensorgram window Copy Graph Biacore Word Pad Paint Word Pad lg Adjusted Sen
21. 60 O a5 B S B 8Bila 2 FRP CRUS VN i EEBOIBNICIAU x o o iB J
22. Copy Custom Methods Methods Rm TY Xd Aldehyde x Surface thiol Method name Copy of Amine Command Solution Contact Time s Flow Rate l min P PRE CONC Specified in Immobilization Setup 6 EDCNHSINJECT EDC NHS 50 50 A WASH Ethanolamine L LIGANDINXECT Specified in Immobilization Setup 4 INECT Ethanolamine Method Method name Edit Custom Methods Copy of Amine 0 OE of Amine Ke Surface thiol Method name Copy of Amine Command Solution Contact Time s Flow Rate l min 79 PRE CONC Specified in Immobilization Setup 5 EDCNHSINJECT EDC NHS 50 50 A wash Ethanolamine D LIGANDINJECT Specified in Immobilization Setup INJECT Ethanolamine OK Set up Method BiacoreeX100 114 5
23. S S Biqcore X100 III Rmax 100RU Rmax x
24. pH Preconcentration binding is too slow Target Reached Preconcentration binding is too sow pH Biacore X100 3 41 3 6 1 Wizards Immobilization Ot
25. A Slop A Slop vs C B A A Conc udgml A A A A A B B A A A BiacoreeX100 1 1 1 1 1 1 1 1 gt gt gt gt
26. N A CV Calc Conc Calib Curve Finish Evaluaqtion Explorer n x Remove Edit e Sensorgram nd AI sensorgrams Binding stability E Report Point Table Biqcore X100 98 5 5 5 1 Create Assay Workflow Binding Analysis Create Assay Workflow Binding Analysis Ligand details Preview of recommended Assay Workflow Ligand name My ligand is Ligand details Ligand name My ligand is a biomolecule with a tag an antibod another protein peas unu gnngthinn else Continue Cancel Binding Analysis Create Assay Workflow Binding Analysis Ligand details Preview of recommended amp ssay workflow Ligand name Protein My ligand i
27. Next gt Start Sqve amp Uv IFD AE RASLE LENDSI Ctrl Break 3 2 30 i 3 Fc Tme Wedow Rep SO LRSD Sepe FRmwReop Base hkh 2 570 5 26y64 03 019 015 00 Yes 70 2 2 2 27113 045 0409 24 3349 No 1 Biacore X100 60 3 Regeneration Scouting Results Display 1stcycle Conditions BP Sample Response Baseline Result Standby Trend Chart Response Baseline Display Conditions Sensorgrqms Display Cycles BiacoreeX100 1st cycle 1
28. TE TRAINING amp Blacore X 100 Control Safi G FE Om Mabini Tr oir E Bee EE PM JE Da 3 FM Tg emu FPP 55 2007 2 756 DS PM w yd El Tlagenarabon Scouting 5 0297 1 90 08 M DURET AHM T ui aa an Geanina aj res pma E Pagm Londions Sampie SM 7007 T DGGES IPM YIAN AFM Tmeng 1 B abina 5 5 2007 112328 AM DANG T2 352PM Trang Pesos Foster mig Tha Suppr Haviegaz bo the ruht of tha main pope provicas context santino mppot ab a i hair x Washim pH Senna 552007 11 1717 OT 102 Tarn Pasa El marbi gH aang a AT 1 LT ATAM Tg 1 E inrchlizalas eH Scouting LT TLTT38 ET TO ESO AME Targ Hesta Er Ea A min iiic Tg TO AA Tis 7 i Frasin rdO e A AME ling A T L etenim reoereraten corten Sra HSEERAM Taning v d L 8 2007 847 21 AM t 4 2007 52820 AM SAT AAM f 5 5 2007 5 22 23 AM LEX ZI3TJ3IEM Th DETIT 22332 FM F GT a FM Tr Bir Aer PM DETIT 201 44 FM www DETO 1 58 07 FM Sauer TA PM Biacore X100
29. Rmax RU Rmax Rmax Rmax Biacore X100 3 81 3 16 F Kinetics Affinity Fit Kinetics Create Add Fit Current Fits Model 1 1 Binding v HEBETT Description Quality Control Report Residuals Parameters 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 1 641E 7 1 000E 9 30 00 5 100E4 7 0 1585 3
30. Enhancement lt Biqcore X100 Next gt Concentration Analysis System Preparation Prime Prime before run Conditioning Startup Run startup cycles Solution Number of cycles 5 Prime Prime before run Prime Startup Run startup cycles 3 Next gt Concentration Analysis Injection Parameters Sample Contact time s First regeneration Contact time s Stabilization period 60 s Injection Parameters Sqmple Contact time First regeneration Solution Contact time Biacore X100 90 4 Stabilization period
31. D 2 VII OK BiacoreeX100 118 5 Cancel Inject Load Samples 3 Hack Unlocked Rack Unlocked Insert the rack and click OK OK Injection command Biqcore X100 5 119 ES admin Biacore X100 Control Software anual Run 1027 2008 2 14 43 FA Ee Ed Wem Commands Bus Tooh eb sendet Ka heb F Lye 1 wm i 5ubiracted Fca TEFA iz Y Tman w ndew Ahes 5 1285 MIA 2 EET Lack scale BiacoreeX100 120 5 BERO
32. 1st cycle 1 Er Hige Cond ons E me Cr 131 Next gt Close Regeneration Scouting Save Settings v Use these settings in the workflow steps T IY J Cs 2 Stabilization period D 4 First regeneration Solution Contact time s Second regeneration Solution Contact time s Save Assay Workflow ProteinA_m IgG Ligand Attachment Overviow Type of assay Drrc hin nz wu Selected chip CMS v ligand name Fictmin d sgana Sensor Surface Preparation Find lewmnobilizzton pH mm irabon buller 10 mid Acetate pH 5 Overview Eg iz esse t En
33. save Ctrl Break 3 2 30 IE m m ES TRAIHING lacere X100 Control Software Assay Conditions Sample 5 11 2007 1 20 48 PM Gi dr wWew Hun Tok Pep fa i J Cyl 1 w Dune Lack scale reed reads 160 pl reed reads 160 pl Di k T a T T FJEETEEFEgQgEFEEE Ordine COME Temperature 25 30 9C Running Ms dy conditions samphe f Assay Conditions Sample Results mouse IgG Relative Response binding 5 stability 5 Conc pg ml RelResp RU binding mouse IgG stability mouse IgG binding 2 mouse IgG stability 2 mouse IgG binding 3 mouse IgG stability 3 mouse IgG binding 4 mouse IgG Biqcore X100 56 3 Results Standby Next gt hE Assay Conditions Sample Save Settings Use these settings in the workflow steps Sample Name mouse IgG Contact time 60 s Dissociation time 130 s Max concentration
34. 3 4 36 Immobilization Result Close Close KW TRAINING Biacore X100 Control Software File View Tools Help RS ME N Assay Workflow ProteinA_m IgG Ligand Attachment Overview Type of assay Direct binding analyte Selected chip CM5 B Ligand name ProteinA ligand Sensor Surface Preparation Overview Results reference Tivat Immobilization pH Scouting LE Find lmwmobakzalson pH Immobilization buffer 10 mM Acetate pH 5 x4 5112007 10 12 37 AM Edit Assay Workflow All results LA Run to foa o LA Run to foa o to find out Erter known vatues known Erter known values i Method in fc 2 Ami Immobilization 5 11 2007 1 Immobili e in fc ine mot s x i Immobilized in fc 2 451 3 RU 11 50 13 AM l Ba Ba Chip id 070511 0939 All results un Results reference Find Sample Conditions Find Regeneration Conditions Run Kinetics Affinity Assay Run S sss Online COM1 Temperature 25 00 oc Sensor chip CM5 Running standby remaining time 4 0 days Friday May 11 2007 Start os WE TRAINING Biacore immo 5 Paint fw first gc Immobilize EJ BAZ Overview Z
35. Sensor Chip Maintenance 9 New chip O Reuse chip Mew chip Chip type CMS V Chip id 070510 1014 Chekti Chip type C Maintenance Dock Chip 2U v 2d Dock 2955 5 d c El Standby l Tools Prime E e Shutdown Biacore X100 Evaluation Software Prime Standby BiacoreeX100 138 6 6 2 Tools More Tools l Tools Maintenance Tools s Desorb Desorb and Sanitize B E Test Tools System Check and Pump Calibration B Service Tools EE Software Problem Report J Flow System Wash Superdean hg Clean Needle and Needle Wash Station Calibrate Needle a Replace Needle pa Replace Peristaltic Pump Tube Cami This applicationprovides a standard farmulafor reparting software malfunction to Biacore Last run time N A Help zm Cancel Tools Biqcore X100
36. n Baseline ld 5 598026 258 023 138 227210 binding 1 2120 5 368737 016 014 0 05 1 93 H stability 1 Id baseline 1 10 binding _1 5 stability _1 10 baseline 1 RU 0 CD RU RelResp 0 0 ZB binding 1 stability 1 RelResp baseline_ 1 7225018 RU 2 boseline_2 binding_2 stability 2 RelResp baseline 2 RU Biqcore X100 5 368815 009 010 000 baseline 1 2 21 Reference line View Reference Line RE
37. Support Navigator Standby amp lt Y Biqcore X100 18 2 Cancel Inject Load Samples 3 Hack Unlocked Rack Unlocked Insert the rack and click OK
38. pH 0 5 2 pH pH 3 5 10 mM pH SA 100 ug ml 10 mM Borate 1 M NaCI pH 8 5 NHS pH 8 5 DMSO Instrument handbook Biqcore X100 VI C 2 pH C
39. biacore Windows LED 22727 power temperature 1 1 2 2 1 200 ml BERT a p YII MEN ou _ m er L BRNA 1 NEM Biqco
40. XD WSJAZSSUAS sSLL CES rine nad o Pae aA AE I i E IH IC fees D 9 Di on o m EILAN ORAAL 91 5 3U 977 0u 3 JALAN OHE HITA Ab IC
41. Standby EP Y D Biqcore X100 5 117 23 24 FORD 2 1 3 Hep TRAINING Biacore X100 Contr I File Edit View Commands Run Tc m 43 EB Cycle 1 2 Heb Vialwell position Il Contact time s Vial well position Inject Val position 11 a Minimum required volume in vial For thie injection 100 pd Contact time 60 120 M
42. Sensorgram window Ll95rY Tools Sensorgram Adjustment of A Report Point time 0 Q Injection Event time 0 Y Adjustment i O Report Point response 0 Injection Event response 0 X Adjustment IZ Report point time 0 baseline X Adjustment C Off A Report Point ime H Injection Event C aimee V Adjustment Report point response 0 baseline __9K Biqcore X100 8 153 8 3 4 amp Zoom Lock Adjusted sensorgram RU 1800 1600 1400 1200 1000 5 Beta2micra high2 amp dE a 800 Beta2micro high1 Beta2micro lovv2 600 Beta2micra Inwy1 400 200 0 200 150 200 250 300 350 400 s 50 50 100 Sensorgram window Time 0 baseline we Caption Cut Copy Graph Export Curves Legend
43. Biacore X100 Evaluation Software What do you want to do Exatuste kinetics affnit Evaluate binding analrsis Sensorgram window This window is displayed autormaticaly when you open data for evaluation Click the Sensorgram button to create an adGNonal S entorgram window Useme 5ensorgram window to examine me s ensorgrams for interaction characteristics this stage wil not appear in subsequent evaluations Using he ensorgram wingow Screen regions and menus Biqcore X100 5 135 BarChat admin Biacore X100 Evaluation Software Multi cycle kinetics Bar Chart m File View Evaluation Tools Window Help 9 kd 63 Sensemram Plot mi BarChat A3 Kinetics Affinity A Concentration Analysis Bar Chart x Remove F Sensorgram AJ ll sensorgrams e Plot Baseline Binding level 1 Binding stability Binding to reference Bar Chart i Bar Chart E gt Report Point Table mm Report Point T able E Am 3 3 o wn h co x E m vo a Cycle Number manam Cycle Purpose Sample Name Startup Sample i EN binding Sample D stability Sample Sample Sample Sample Sample Sample DO e XO C MN Response Tools v Relative
44. E Cycle Pupose Ovetays v P Cycle lt Overtay gt JJ ozs cta d 2 cded odes Ode Prose Sampie ame cone MM Yes _ 1istartup Buffer LEES sample Bim micro low ing cc cie ims Ctrl BiacoreeX100 152 8 8 3 2 Sensorgram window Tools Color By Sample Tools gt Report Point gt Id and Marker 8 3 3
45. standby Biacore X100 3 53 3 4 2 Assay Find Sample Conditions pM Find Regeneration Conditions cm Run Kinetics Afhinity Assay Find Sample Conditions Run to find out 2U v 2d S BE Enter known values Assay Conditions Sample Injection Sequence 12 Reference subtraction Chip Flow cell Chip type 5 Injections in analysis cycle Flow Cell1 1 2
46. BiacoreeX100 3 7 1 admin Biacore X100 Evaluation Software Single cycle kinetics 2 analytes Fie view Evaluation Tools Windaw Help ue A Sensogam Plot 25 Bar Chart A Kinetics Affinity A Concentration Analysis g x Remove Edit PA All sensorgranns es Curve Name Fc 2 1 P Cycle Purpose Overlay e Cycle Overlay e ensorgram sJ All sensorgrams RU Sensorgram P Plot 450 Zoom Lock Baseline Report Point Table m Report Point Table 1500 3000 3500 Thursday October 30 2008 Biacore X100 Evaluation Software A Kinetics Afinity 3 10 72 Biacore X100 72 3 3 10
47. IX D 2 1 1 7324 FID Ld Kp M 1 Kp KA 1 M A BSAB
48. Biacore X100 3 31 i 75356 A SU Ae IO LASO Eme eey ee 3o SE Do in XLI HE Tea 3 Zb 11 gu IE Fe 1 l Results PE Immobilization pH Scouting Results ProteinA 10 mM Acetate 5 5 10 mM Acetate 5 10 mM Acetate 4 5 10 mM Acetate 4 Relative Response 2 10 mM Acetate 5 5 ProteinA 2 10 mM Acetate 5 ProteinA 2 10 mM Acetate 4 5 ProteinA 2 10 mM Acetate 4 ProteinA pH pH5 0 BiacoreeX100 3 3 pH pH pH4 pH NHS
49. Results reference Jg Biacore X100 Aim for immobilized level NHS quu Chip CM5 Ligand Response Flow cell Procedure Method Final RW 1 Targetlevel Amine Protein N A No Preconcentration binding is too fast Preconcentration binding is too fast Target Reached Preconcentration binding is too fast
50. Q O O Quality Control Report Residuals Parameters o Kinetic constants are within instrument specificatians o Kinetic constants appear to be uniquely determined o Mo significant bulk contributions RT Found eo Go o Check that sensaragrams have sufficient curvature o Examine the residual plot Pay attention to systematic and non random deviations BiacoreeX100 3 77 AN V 2858 a 105 107 1 Ms ka 105 0 5 1 s ka kd Rmax ORDRE R RI
51. TEL 03 5331 9336 ERIT 9 00 17 30 WWwW gelifesciences co Jp e mail Web 2009 GE GE
52. Find Regeneration Step TTE Next gt BiacoreeX100 124 5 ssay Conditions Sample System Preparation Prime v Prime before run Conditioning COTIGTHOTHTRI Cy CIE Prime before run Prime Next gt ssay Conditions Sample Injection Parameters Sample is Contacttime 0 _ Me 120 180 WE Concentration unit pg ml sample Concentration pg ml M nM oa im amiami miata mama 50 mouse IgG Sample Position 1 50 mouse IgG Sample 50 mouse IgG Sample 50 mouse IgG Sample Full Deionized water Water Rack Positions Next gt Biqcore X100 5 7 amp O 88 0 125
53. Keyword Table Tools Keyword Table Keyword Table Conc ugiml MW Da Reset All Filters 1i Startup 2 Startup 3 Startup 1 Add Keyword 4 Startup 5 Startup Rename Keyw 6 Sample 7i Sample 8 Sample 9 Sample 10 Sample 11i Sample 12 Sample 13 Sample 14 Sample 2 0 ug ml 15 Sample 16 Sample 17i Sample 18 Sample Remove Keyword Concentration Unit Biqcore X100 E Concentration Analysis Create Calibration Samples Calibration Curve Settings How Cell Fc 2 i v Report Point stability Calibration Table Calibration Curve Curve 1 RU 5000 0 3178 0 6428 Relative Response 0 6418 1 263 1 262 0 08591 Rhi 1 11E 04 Rlo 8547 Al 25 3 A2 1 052 E 5 Chi 75 48 Response Type Relative Response v Fitting Function 4 Parameter w we m IgG 10 15 20 25 Concentration pg ml Calibration Curve Settings Flow cell Report Point stability
54. Add Report point E Edit gt Report point D Add Add Report Point E x Report Paint 1d Time 281 0 3 windo B Z O s M Baseline v Apply to all curves in this cycle Hep Canes ld Baseline 0 Biqcore X100 22 2 2 1 3 End manual run Run Stop Run PL Biacore X100 v This will end the manual run when the last queued command is completed Cancel OK standby Biacore X100 2 23 2 2 File Save eo 2 3 File
55. Rack Position Next gt Start Biacore X100 A Changes detected in wizard template Save As Don t Save l Sqve 3 2 30 Standby Biacore X100 3 43 Methods Copy Custom Methods Methods Rm TY Xd Aldehyde x Surface thiol Method nam
56. 169 0073 3 25 1 TEL 03 5331 9336 FAX 03 5331 9370 e mail Tech JPGge com ISO 9001 2000
57. Sample 4 Find Regeneration Step Next gt Biqcore X100 54 3 Assay Conditions Sample system Preparation Prime v Prime before run Conditioning Prime before run Prime Next gt ssay Conditions Sample Injection Parameters Sample Name Contacttime 60 Ms GW 120 180 Concentration unit pg ml sample Concentration pg ml M nM MNT ERE 50 mouse IgG Sample 50 mouse IgG Sample 50 mouse IgG Sample 50 mouse IgG Sample Full Deionized water Water Rack Positions Next gt BiacoreeX100 3 55 Start
58. 30 60 ul min 3 3 57 10 s 10 30 Biacore X100 XI D 3 ESL LELAS A 450 3 B B C A i RU WE i A s RU vs C A
59. Er Hige Cond ons E me Cr 3 61 Next gt Close Regeneration Scouting Save Settings Use these settings in the workflow steps Stabilization period D 4 First regeneration Solution 77 Second regeneration Solution Contact time s Save Assay Workflow ProteinA_m IgG Ligand Attachment Overviow Type of assay Drrc hin nz wu Selected chip CMS v ligand name Fictmin d sgana Sensor Surface Preparation Fiod levmobikraton pl Eg iz esse t Entar known oc ge Find Sample Conditions kd ig Puto fec c Erter roue values J Find Regenerabon Condi ons m Run 15 fird aa Erim kroan valusa j Senso
60. Oracle Database DOS amp Stop Database FERSTELE TREE ERO LIT RES REM SE S 9 38 TITRE EGRE LITE EET UTR DTE E d The racle ServiceA E service is not starte More help is available by typing MET HELPHSG 3521 oraclexe sapp oracle product 1H 2 BH zervuer BIN Oracle Database 64383 BE Stop Database Biqacore X100 148 7 7 2 4 50 ml HBS EP
61. xS Da RU Da S 50 000 Da 1 000 RU 1 20 000 Da Rmax 20 000 x 1 000 50 000 x 1 400 RU 10 000RU RUvsC ka kd KD RN lE U P21 Fo1RIGZ23 2278 0RE RU RU 200x1 Sx RU 1000 x 1 S x S 50 kDa
62. 5 4 5 4 1 Other options Manual Run Manual Run Flow F Flow rate EI v imin Flom path Q Flow path 1 Q fed Flow path 2 Reference subtraction E Flow path 1 2 30 ul min Flow path 1 2 Reference subtraction 2 1 Start Save Biacore X100 5 115 admin Biacore X100 Control Software Manual Run 10 27 2008 2 14 43 PM i 12 jep Li 10000 5 10 15 Fel Time Window AbsResp SD LRSD Slope RelResp Baseline ld E Keywords in cycle 1 Value l Flow 30 F
63. Cancel Inject Load Samples 3 Hack Unlocked Rack Unlocked Insert the rack and click OK OK Injection command BiacoreeX100 3 49 EZ admin Biacore X100 Control Software Manual Run 10 27 2008 2 14 43 PR EG Ed em Cg Maa fedh pep sendet Ka heb Ep 1 wm Dune 5ubiracted Fr TEFA iz Y Tma Www AhaRiesn 5 1285 MIA 2 EET Lack scale Biqcore X100 50 3 BEROR Regeneraqtion Command Regeneration Wal position lg UK Donate 8 Minimum required volume in vial for thie injection Z0 pd Vlll l admin Biacore
64. Sensor Surface Preparation Find Immobilizahon pH Immobilize Find Immobilization pH Run to find out amp 2 U v 2d Enter known values Immobilization pH Scouting Setup Detection REDRA Sr TR CD pH Buffer Name 10 mM Acetate 10 mM Acetate 10 mM Acetate 10 mM Acetate fBTAR S5 d5 d O TRIBIAROD pH Next gt Biqcore X100 5 103 Immobilization pH Scouting Injection Parameters Prime Prime before run Prime Ligand Name Contact time s 60 Surface regeneration This surface wash will be run once at the end of each cycle 50 mM NaOH Next gt ES Immobilization pH Scouting Rack Positions 45 ProteinA Gioia 10 mM Acetate 5 45 ProteinA Sample 10 mM Acetate 4 5 45 ProteinA Sample 10 mM Acetate 4 135 50mM
65. All results BiacoreeX100 108 5 5 3 C lI Immobilize immobilization Setup ee Prime before run Prime Flow cell 1 Method Ligand solution Amine 2 Ez Flow call 2 E Immabilize tow cell 2 Method Amine v Aim for immobilized level Ligand solution Protein in 10 mh Acetate pH 5 Specify contact time Contact time z 420 1 2 7 Next gt Immobilization Rack Positions 126 Ethanolamine 75 Protein Immobilization 85 EDC Mix Fc 2 85 NHS Mix Fc 2 Empty EDC NHS min capacity 13 Ll Mix Fc 2 Full Deionized water Water
66. EDC NHS 10 ml 200yul 11 mm 20 1 4 200 u BiacoreeX100 V 1 BSA 10 mM 10 mM Borate 1 M NaCl pH 8 5 5 200 ug ml 10 mM
67. f Start Save amp 2U v 2d S MEERA LEDET Ctrl Break 5 2 104 VE TRAINING Biacore X100 Control Software Assay Conditions Sample 5 11 2007 1 20 48 PM EZ Edt Vew Run Took Heb Bie ree wo Cuve Sensorersm Fes RU L Lock scole 30000 pined Start 7082 bined Star Pos3 T i i ji EET T 9 8 fa ONE NONI iim v Somoe Sampie Sample Sorcte Sample Sorpe Sarpe Sample Sane Sample Sample mm i Ranning 5S by Conditions sample V Assay Conditions Sample Results mouse IgG Relative Response binding 5 siability 5 binding mouse IgG stability mouse IgG binding 2 mouse IgG stability 2 mouse IgG binding 3 mouse IgG stability 3 mouse IgG binding 4 mouse IgG BiacoreeX100 126 5 Results Standby Next gt Assay Conditions Sample Save Settings Use these settings in the workflow steps Samp
68. Next gt Concentration Analysis Calibration Curve Calibration Curve m Calibration points Calibration Curve Calibration Curve Analyte name Calibration points Concentration wasp 6 Next gt Biqcore X100 4 91 Concentration Analysis samples Sample table samplei 1000 sample 500 sample3 200 sample4 50 samplei 1000 sample2 500 sample3 200 sample4 5p Samples Sample table Sample id Dilution factor PX24bSSMB
69. Run gt Stop Run Biacore X100 A This will stop the run gm Ceen we Stop Run Run Stopped Finishing current cycle please wait Abort cycle by CtrlIBreakl Standby Ctrl Break Biacore X100 v You have aborted the run Do you want tn wash the system Recommended Yes Biacore X100 4 93 Standby Biacore X100 Evaluation Software Biacore X100 94 4 Biacore X100 Evaluation Software 7 Concentration Analysis 2c 7 w 2d 2 4 2 WE 4 2
70. Regeneration parameters 3 Solutions 1 Contact time Solutions Contact time 7 VIlll Number of cycles for each conditions Settings Regeneration solution Contact time s 2 2 Next gt Biqcore X100 5 Bae RD Regeneration solution1 2 Contact times 1 5 129 V Regeneration Scouting Rack Positions 105 10mM Gly HCl
71. 100 kDa 1 200 x 1 1 x 50 000 100 000 100RU 1000 x 1 1 x 50 000 100 000 500 RU 100 500RU Biqcore X100 IV C 1 N g CM NHS N NHS CONH COO COO CONHCH CH OH ONKS RU BR 1000 50 EDC N ethylI N 3 dimethylaminopropyl carbodiimide hydrochloride NHS N hydroxysuccinimide 1 M ethanolamine hydrochloride pH 8 5
72. EDC NHS Next gt Start Save Ctrl Break 3 2 30 TPE Immaobilirarion 5 11 2047 11 50 13 AM ES 4 A E EH A F E Crea m Cor Larzrriram Fea E p A FraoteinA E Lex cle AU EDC NHS amp B 2r 7 VN ie uu Aim for immobilized level Standby RESNA Immobilization Results Biqcore X100 Xe Immobilization Results Ligand Rasponse Final RU Target Reached Targetlevel Amine ProteinA 3 3 451 3 Yes I Response Bound Response Final RU
73. 8 1 1 2 2 Tools Prime E e Shutdown Biacore X100 Evaluation Software Prime Place a bottle with buffer on the left hand tray and insert the inlet tubes e Stqart Prime Prime Priming please wait Time left 5 56 Prime The Prime procedure is completed Close Standby 1 7 Prime Biqcore X100 1 2 3 Tools Set Temperature Tools Help 1 Prime Shutdown Biacore X100 Evaluation Software Stop Standby Load Samples wv Rack Illumination Undock Chip Set Temperature Change Password More Tools
74. Immobilize ligand covalently using Sensor Chip CM5 Create Assay Workflow Kinetics Affinity Choose ligand attachment approach Preview of recommended Assay Workflow Ligand details Ligand name Sensor Surface Preparation My ligand is Ligand attachment approach Recommended Q Capture using own antibody and Sensor Chip CM5 Immobilize ligand covalently using Sensor Chip CM5 Ligand attachment overview Type of assay Direct binding ds Selected chip CM5 4 Ligand attachment overview Preview of recommended Assay Workflow DRN Continue Save Assay Workflow As Save in folder Folder B nakayama E Users Name Modified 2 8 Training ProteinA mlgG2 5 9 2007 2 26 05 PM lg Group IJ ProteinA mlgG 5 9 2007 9 58 30 AM By murain iJ 070509 5 9 2007 9 27 23 AM EJ2 5 8 2007 2 31 33 PM Bg Sheny Yoshimi Ej5 5 8 2007 2 29 32 PM BR Sotwareexercses 4 5 8 2007 2 08 54 PM Bl system check Ej 1 5 8 2007 2 08 04 PM administrator Instrument Service
75. Standby 7 7 1 1 Standby Standby 7 200 ml Online COMI Temperature 25 00 c Sensor chip CMS Running standby remaining time 7 0 days 7 1 2 Tools Undock Chip b Biacore X100 This will undock the sensor chip Undock Chip Sensor chip Sensor Chip Maintenance Biacore X100 7 145 Dock Chip opi om giao mio
76. Fit BiacoreeX100 83 Current Fits Delete OK i Current Fits Biqcore X100 84 3 3 7 2 SER admin Biacore X100 Evaluation Software AAL_LNFP1 File View Evaluation Tools Window Help 9 kd ue Sensorgram Plot mi BarChatt 2 Kinetics Affinity A Concentration Analysis n W All sensnrgrams mB TE Curve Name Fc 2 1 e Cycle Purpose lt Hyerlaw gt Cycle Overlay ensorgram Sensorgram a 5 sensorgrams RU g IF Zoom Lock Baseline Binding level Binding stability Binding to reference E Report Point T able mm Report Point T able ix Remove Edit Sample Startup 3 10 72
77. OK Injection command Biqcore X100 2 19 Injection command l 2 2 E Biqcore X100 2 1 2 RU 2 3 Jg
78. B A B amp AB AB B AB2 Heterogeneous Analyte A1 B amp A1B A24B A B Heterogeneous Ligand P Xx24FlewucaxttosGe owoissu ut isoJHFbpb z24FHr8531 A B1 amp AB1 A B2 AB2 Two state Reaction conformation change A B AB AB Biqcore X100 76 3 Ft Kinetics Affinity Fit Kinetics Create Add Fit Current Fits Model 1 1 Binding v 1 1 1 Binding Description Quality Control Report Residuals Parameters 1 1 Binding Quality Control 7 tE 3 14 Quality Control 5
79. C 90 CM5 N se CM NHS N NHS S S
80. pH4 pH NHS NHS pH8 5 pH pH pH5 Immobilization pH Scouting RU 100ug ml Immobilization pH Scouting pH Next gt Save Settings immobilization pH Scouting Save Settings F3 m Use these settings in the workflow steps Buffer Commeni Buffer name pH E Ooo d fill 10 m4
81. Kp A B AB Biacore Ko ka ka k Ms k 1 ka A B AB Ko kal ka Ka Kal ka Biqcore X100 X Ko kaka R ka Rmax C N 4 Req
82. Selected Capture1 Sqmple1 Settings for Capture 1 Flow path Second Custom Assay Wizard Cycle Definition n Settings for Capture 1 Available E Capture solution capture varies by cycle Sample Sample 1 Contact time amp 0 s varies by cycle Enhancement Regeneration Flow rate 5 ul min Extra wash after injection with Stabilization period 0 Next gt Custom Assay Wizard Report Points Rame sec SeoreiAter saregewdor m baseline 10 Before Start of 1 Concentration 1 m binding 1 10 Before End of Sari 1 Concentration 1 stability 31 16 After End of Sample 1 Concentration 1 E binding 2 10 Before End of Sample 1 Concentration 2 16 After End of Sample 1 Concentration 2 10 Before End of Sample 1 Concentration 3 15 After End of Sample 1 Concentration 3 10 Before End af Sample 1 Concentration 4 Ar ALL p GE p Report points Next gt BiacoreeX100 3
83. AbsResp Binding level Baseline RelResp Binding stability Bqseline 50 RelResp Binding to reference Binding level El Report Point Tabe Work qreq Evaluation Explorer Biqcore X100 8 151 8 3 Evaluation explorer Sensorgram All sensorgrams amp Work area Sensorgram window 8 3 1 Sensorgram window Curve Name Fc 2 1 m H 4 Cycle Purose Overays 7 JP 4 Cycle Overtay h 6 Curve Name Fc 2 1 x Joy lt U eia nd 2Uv2L BSO20 t r Ctrl
84. Add User Password never expires Add Biacore X100 9 161 dd User Uzer name Full name Group Description User password Password User cannot change password Password never expires User must change password at next login User account Account is locked Account is disabled User name Full name Group Administrators 7e d Users Description User password Password L User cannot change password Password never expires L User must change password at next login XZrIvV2 amp AdTe
85. 3 Solutions 1 Contact time Solutions Contact time B 7 VIl Number of cycles for each conditions Settings Regeneration solution Contact time s 2 2 Next gt BiacoreeX100 5 Bae RD Regeneration solution1 2 Contact times 1 3 59 V Regeneration Scouting Rack Positions 105 10mM Gly HCl pH3 0 Regeneration 135 10mM Gly HCl pH2 0 Regeneration Full Deionized water Water Rack Positions
86. Keyword Table Tools Keyword Table Cycle Cycle Purpose Sample 1i Startup 2 Startup 3 Startup 4 Startup 5 Startup 6 Sample 7i Sample 8 Sample 9 Sample 10 Sample 11i Sample 12 Sample 13 Sample 14 Sample 15 Sample 16 Sample 17i Sample 18 Sample gt Keyword Table Conc ug ml MW Da Reset All Filters Concentration Unit ug ml BECL JYYVYILT ZPND Biacore X100 3 73 Kinetics 7 Affinity Select Curves Create Curves Sample 489 O w Temperature 25 i Cure Fc 2 1 Cyce Conc Flow ir Time ii ER D nM T B Zoom lack Show concentration series Show blanks Show average blank z sample 3 11 3 11
87. Standby Standby 3 4 ee Biacore X100 140 6 6 2 2 Desorb and Sanitize 1 1 1 20 C Maintenance Tools Desorb and Sanitize Start l 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 Next gt Desorb and Sanitize Required solutions from Maintenance Kit BlAdesorb solution 1 about 10 ml BlAdesorb solution 2 about 10 ml BlAdisinfectant solution about 15 ml Next gt ATW 1 Desorb and Sanitize Step 1 Place 10 ml Bl amp desorb Solution 1 on the left hand tray and insert the pump inlet tubes
88. 132 NN 4e Le nar zio AT CRECEN 156 6 1 kk 137 6 2 Saaliif 138 6 2 1 B 1 1 a o 2 139 6 2 2 Desorb and Sanitize ec 140 6 3 142 20 EE 144 7 1 kk 144 7 1 1 Standby 41 1AIIIIELEILIL ttt ttt ttttte itte ttt tatto ati0 144 7 1 2 Eea E 144 UN E APIS TEE 148 8 149 8 1 kk 149 Biacore X100 8 2 ecc 149 8 3 2 RE ER 151 8 3 1 EER 151 8 3 2 ec 152 8 3 3 2 152 8 3 4 entente 153 8 4 i B A 0 D 1 154 8 5 kk 156 8 6 JI 157 B eU uestes buch i 158 Manolo 159 Erie iC RN C 159 9 2 ku kk 162 Biacore X100
89. 4 3 Response Type Relative Response Fitting Function 4 Parameter Linear 4 Parameter Calibration Table Curve Cycles Conc Response RU pegs Calc Conc CV Calc Conc CDZe 8o p Biacore X100 4 3 Report Point stability 20 stability Custom Report Points Add 27v 2d S f Add Custom Report Point Repat paint OOOO Window 5 wt Position the report point 10 seconds afte Calculate response relative to report point Cycles Apply To Selected Cycle P
90. Req 18 RU Req 187320 v F Biqcore X100 86 3 Next gt Ea Kinetics Affinity Fit Affinity Create Add Fit Model Steady State Affinity cS 6e 4 Concentration Fit Affinity Model Steady State Affinity Fit Ea Kinetics Affinity Fit Affinity Create Add Fit Current Fits Model Steady State Affinity EE Lc oes 6e 4 Concentration bon on ee Report Parameters KD M Rmax RU offset RU Chi RU G Report Kp M Rmax RU Biqcore X100 3 87 offset RU bulk effect Chi RU G Parameters 9 7 Finish Evaluation Explorer Kinetics Affinity
91. 1 Wizards Immobilization Otheroptions iens Zj Wizards gt Immobilization New Immobilization Setup gt Chiptype SLE v How cell 1 O immobitize fow cett 1 Flow cell 2 O z immobitize fow cell 2 Hp Commeres l Prime before run Method Capturing molecule ligand solution Target level RU Wash solution Method Capturing molecule ligand solution Target level RU Wash solution Chip type Method Aim for immobilized level Wash solution Specify contact time OBlank immobilization Next gt Surface thiol Target level 50 mM NaOH
92. Commands 1 Selected Available Add gt 3 9 68 Setting for Sample 1 Single cycle kinetics Concentration per cycle 1 5 ERIS e CI N 5 Contact time PXx24 4 NJ GD 8120 180 Dissociation time REDE 120 180 Flow rate ul min 30 l min Flow path Both 12 Extra wash after injection with PX24 ANDUOOXUSODUUS112e 2w 3 amp l U0 tJU Biqcore X100 68 3 3 9 Cycle definition Commands Available Capture Add gt
93. Saqve Biacore X100 3 27 NE TRAINING Biacore X100 Control Software File View Tools Window Help Assay Workflow ProteinA m IgG i aehrneni Overview Type of assay Direct binding Selected chip CM5 Ligand name ProteinA Edit Assay Workflow Sensor Surface Preparation Overview Results reference Find Immobilization pH J2zFl biE boEESS Immobilize SS m Assay Overview Results reference Find Sample Conditions PT2FHBREOElHSS Find Regeneration Conditions BES RORIHRE ESINEORS Online COM1 Temperature 25 00 9C Sensor chip CM5 Running standby remaining time 4 0 days sf Start E V8 TRAINING G Biacore ProteinA 5 Paint fl My Pictures Run to find out L Id Run Overview Results reference Enter known values Overview Biacore X100 28 3
94. BiaqcoreX100 Evaluation Software Evaluoation 8 1 Windows Start Programs All Programs Biacore Biacore X100 Evaluation Software 8 2 I 5 File Open OK BiacoreeX100 150 8 ml rp am wj JU N z Cur DETE mia Eom Cim Pirpana cmn he EE EE d B mpra nu as i g m L i hj i naa 1 Sensorqram Window Evaluation explorer me Evaluation Explorer le Sensorgram I Plot Baseline
95. NHS pH8 5 pH pH pH5 RJS Immobilization pH Scouting RU Ql 100ug ml Immobilization pH Scouting pH Next gt Save Settings Immobilization pH Scouting Save Settings E setere treo intents Buffer Buffer name Ho T pH Save BiacoreeX100 TRAINING Biacore X100 Control Software Fille View Tools Help Assay Workflow
96. l FS Kinetics Affinity Select Curves Create Curves Sample NFP 1 v Temperature Curve Fc 2 1 Flow rueloeee seme o 9 ee v 0 30 500 30 250 30 125 30 62 5 30 31 25 30 30 30 30 Zoom lock Show concentration series Show blanks C Show average blanks Multiple Hmax Adjust Injection Start BiacoreeX100 3 85 3 11 73 Next gt YS Kinetics Affinity Select Data Create Curves CORE Hain a pl min s s 500 30 5 Fc 2 1 5 Fc 2 1 250 30 7 Fc 2 1 125 30 8 Fc 2 1 62 5 30 9 Fc 2 1 31 25 3 10 Fc 2 1 30 11 Fc 2 1 3 12 Fc 2 1 30 13 Fc 2 1 30 v v v v v wv v C Zoom lock 0 Affinity gt YS kinetics Affinity Select Affinity Data Create Settings Calculate response at position 4 seconds before injection stop with window 5 seconds 3e 4 Concentration TE s aai Bek jJ Mew gt J comes RU
97. Biacore Templates 7 v 7d 2 Biqcore X100 66 3 Select Biacore Template Detection Chip Flow cel PE Chip type CM5 Purpose Evaluation purpose Kinetics Affinity he Prime Conditioning Frime before run System Preparation Detection Flow cell 1 2 Reference subtraction 2 1 Chip e e e Chip type Purpose Evaluation purpose Id Kinetics Affinity Prime before run Prime Biacore X100 3 67 Custom Assay Wizard Cycle Definition Commands Settings for Sample 1 Available Selected Solution eE S E E varies by cycle qum Eus Single cvcle kinetics Concentrations per cycle e d Contact time 120 s varies by cycle Dissociation time 800 s varies by cycle Flow rate ao ulmin Elow path Boh vw Estra wash after injection with bo Cycle Definition
98. H2O 4 ml Start Hiri Sysberm Check and Pap Colibratssn SA 71 108 57 AM EL Ti Cancel File name Saqve System Check and Pump Calibration System Check and Pump Calibration Hama Date Run Daie 1M Temperature 2580707 Fila System Check and Furmp cal ecadian 51062007 11 08 51 All T Feristaltic Pump Calibration Pass Raespaonaa Pass Injections Pass Noise Pass System Check and Pump Calibration Late Mersion 3 0 REZ 4 Pass Fail Fail BiacoreeX100 144 7 7 7 1 2 D Standby 7
99. 2 BIAdesorb Solution 17h 10 m Start 8 l 1 2 Biqcore X100 6 141 2 Desorb and Sanitize Step 2 ine 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 m Start 7 l 2 3 ATW 3 Desorb and Sanitize Step 3 Wipe the pump inlet tubes with a moist tissue Place 15 ml diluted Bl amp disinfectant Solution on the left hand tray and insert the pump inlet tubes 4d LBI oDL2 2Z2 9xGuHB d4 7Lwkcai 272 ARLI BlAdisinfectant Solution 1 5 ml 20 m Start 23 l 3 4 RT YV J 4 Desorb and Sanit
100. 30 ul min Flow path Flow path1 1 Flow path2 2 Flow path1 2 1 2 Reference subtraction 2 1 Biqcore X100 14 2 Load Samples Rack Unlocked Rack Unlacked Insert the rack and click OK rack locked OK Manuql Run Start Save Results As Save in folder Users Assay Conditions Sample 4 3 2007 1 33 49 PM 4 24 2007 7 15 mdr Kinetics Affinity 4 3 2007 2 51 26 PM 4 24 2007 7 15 Single cycle kinetics 2 analytes 4 24 2007 7 16 1 Assay Conditions Sample 1 4 24 2007 7 16 Binding Analysis b2u 4 24 2007 7 164 Binding Analysis Sandwich Assay 4 24 2007 7 16 E Concentratio
101. TRAINING Biacore X100 Control Software Create Assay Workflow Other Options kie JE Ji 2 BiacoreX100 Results References Overview
102. IV UL RCRUM lI IV UY uta dm T J vn 19 23 I Ci CG GCC VI 927 dio acd OR 4 25 28 53 65 68 88 92 99 101 102 1235 149 E E REED 9 RI CE X 81 RE VII XI 76 77 79 81 82 DALT A P NT RR RE T E TE IX X XI SE 2 222 71 IX 78 III VII IX X XI 24 78 86 e A eS m oerte Merci reti rendant 29 30 35 38 42 55 59 64 70 92 103 104 109 112 125 129 134 ZA P RR E TM VII Biacore X100 roses ic AES ME RN RR IX 78 dr NN R eEE S Ii Vis L HI HI IV V VI VII XII 6 7 13 25 28 31 32 34 35 36 37 38 39 40 41 43 70 88 99 102 105 106 108 109 110 111 115 156 148 lid A E A E A N E E E E III 32 106 c zh RR RR RR RR ONERE 79 80 81 82 VII VIII IX 79 80 82 L2 m VIII 17 46 50 51 58 59 61 67 89 90 116 120 121 128 129 131 PAS ll ad E E A T RON RUND SERNA ESAE TERRE ER AEEA IIl 80 L Eo 142 NR IX 65 71 wIDAU eA 7 NEPOTUM MEER IIl VII cpu need emu MdL DC II ETC CID M OCT III UOI T D OM C ET LM E DII C ACT EDO E DUIS 76 EIOS IU 77 poU IUUD Eu 137 de eeu 2100 IE TERR BE Sette ieeteniece
103. sensor chip Standby Standby 7 3 Dock Dock Undock sensor chip Gp sensor chip 1 5 Reuse chip Reuses id Dock Chip Chip id D070426 1110 Chiplot no 1170296 l Details Chip Properties Chip id Chip lot no 070426 1110 117
104. 34 3 3 3 C lI Immobilize immobilization Setup ee Prime before run Prime Flow cell 1 Method Amine 7 zz Flow cell E Immabilize tow cell 2 Method E Amine v Aim for immobilized level Ligand solution Poe in 10 mh Acetate pH 5 I Specify contact time Contact time 420 1 2 7 Next gt Immobilization Rack Positions 126 Ethanolamine 75 Protein Immobilization 85 EDC Mix Fc 2 85 NHS Mix Fc 2 Empty EDC NHS min capacity 13 Ll Mix Fc 2 Full Deionized water Water Rack Positions
105. Administrator Users export import Administrators T Administrators O O O O uses gt O Jo JO Users import Biqcore X100 165 A Add RepDoFE OIL o odo Nii EI i 21 AOPBISEEOEOES sec er ono ita E d d edu M MM EE 159 161 162 164 A 24 66 85 86 87 88 Al Ti dor Jdmmobilized leve 2 eni an eet NE i D etii 38 40 41 111 B Dur Cabs eut ud aM E 135 DOSEN S 20 152 BOSGIINE nerenin Y 21 51 60 121 130 150 BIGCORE Tempat oS e fii 65 DINAIG Si nii 20 40 Bilcllhg ADdIVSIS eia i i ii ik 88 98 132 Bivatent Anglyte NS 75 Blank Intmobllizatiohi assis ttes co ie en RRA NR 41 111 Bulk Elect i osos oan Dd d Ec D i E VII C Calibr aton bU Ben ioenos rem omdi tontonn6oEItxosd umbo dindo n 90 95 E i ape PP KC P MeL iO Gi i 79 gel 6 8 36 39 61 110 131 139 141 Concentration Analysis estetica etta insti i 88 CODEC DIT On UA pent Mo vxo e UNDA Gi 16 47 58 67 89 117 128 CODCOD e coire on dedi nuditate de aseo deuda bur pis cu tUe 156 GIHde cet Ed t E
106. Biacore X100 5 123 5 4 2 Assay Find Sample Conditions TEM Find Regeneration Conditions ES a Find Sample Conditions Run to find out 2 Y v 2d S BE Enter known values Assay Conditions Sample Injection Sequence 12 RE Reference subtraction Chip Chip type Injections in analysis cycle Flow Cell 1 Flow Cell 2 Sample 1 Capture 5 1 2
107. ug ml J KD 1 10 10 ug ml Biqcore X100 VI
108. EE 16 2 1 2 ee 20 BL nib NIRE 22 2 2 P 0 23 2 3 kkk 23 3 24 21 i 24 3 2 pH ee 28 3 3 E V L RODIE E a 34 3 4 RISORA ROBER RR eeeeeetenttetteetentnnte 44 3 4 1 eee 44 3 4 2 LL kk 53 3 5 I 62 3 6 DIA Vaa a EI AV AOE E a 65 3 7 71 3 71 DART 1 2 BERT saa dut reati A A i 71 3 7 2 kk 84 dsl 88 4 1 EDADE aE AO S 88 5 ee 98 Biqacore X100 5 1 E E A a E O D F 98 5 2 pH BROR oooeeeneeeesee1esoeseereesssseerrssseeeeeesssseorrsssseetnressnsterrssssenenrsssnees 102 5 3 4k 108 5 4 RISORA ck O Bi ESEUFOD RR eeeeeeeteetteteentettnnnenns 114 5 4 1 aa A V E Eo r E 114 5 4 2 Lz 123 5 5
109. Firad 5 amid erdt m Pin io fd ou Erin imn Biqcore X100 12 1 Open New Wizard Template eed j Concentration Analysis j Kinetics Affinity 4j Binding Analysis Custom Assay Wizard NIN E Biqcore X100 2 13 2 2 1 Other Options Manual Run Manual Run How F Flow rate 10 v ulmin How path Flow path 1 O Flowpath 2 Reference subtraction O Flow path 1 2 Manual Run Flow rate l 5 10 30 ul min 10ul min
110. Biqcore X100 F Tene Wie Ahea 5D LASD Gks Ref Bashin E 5 EM nM 2 ae 5 Velit D Dip du l ag 8 ei 493 Me 5 105 E T in ana kj Keyeec ds rs cele 1 LT bee Lele Sampi bing C ocleT qd DH pigum ie rame 10 h e Spie o aya Par Se 1 Sample Frater Tong m 1 REDKI FAE Results ES Immobilization pH Scouting Results Relative Response 2 10mM Acetate 2 10 mM Acetate 2 10mM Acetate 2 10mM Acetate ProteinA 10 mM Acetate 5 5 10 mM Acetate 5 10 mM Acetate 4 5 10 mM Acetate 4 d Reesp RU 5 5 Protein 5 Protein 4 5 Protein 4 ProteinA pH pH5 0 BiacoreeX100 106 5 E 5 3 pH pH
111. Cut l T Adjusted sangorgram 7 om Lack ax T Ju E 4 T ee M 300 F 2i xut 250 f i H E ox Betazmicro Figh2 Betazmiru Fighi E b j Beta om crn ma a 130 j PE 3 E 5 Bpetazmicru hrw 3 H Tu su di Ej aet u en B Leu n id Eri a i 1 i D i 1 D x i b i 1 Zn 5n 1n 15n 206 5n mn 5n AD Time f hasrdinge Biqacore X100 154 8 8 4 Sensorgram window Scale Auto w Caption Undo Cut Scale Copy Graph Export Curves Gridlines Legend Scale Y Scale Auta Auta Logarithmic Lagarithmic Min Max OK Biqcore X100 A Scale 1 Scale amp uto Auta Logarithmic _ Logarithmic c 8 155 Legend Legend Fosition 2 Hidden 2 Q Top Right C2 Bottom Right
112. 10 SS RI RI RI 0 Constant Biacore X100 80 3 1 ka ka Rmax lt
113. 3 2 pH JHAY KARRO pH C 2 VI Sensor Surface Preparation Sensor Surface Preparation Find Immobilization pH Find Immobilization pH Run to find out 2 Y v 72d Enter known values Immobilization pH Scouting Setup Detection REDRA Sr TR CD pH Buffer Name 10 mM Acetate 10 mM Acetate 10 mM Acetate 10 mM Acetate Next gt BiacoreeX100 3 29 Immobilization pH Scouting Injection Parameters Prime Prime before run Prime Ligand Name Contact time 180 H 60 Aie Surface regeneration This surface wash will be run once at the end of each cycle 50 mM NaOH Next gt Immobilization pH Scouting Rack Positions 45 ProteinA 10 mM Acetate 5 45 ProteinA Sample 10 mM Acetate 4 5
114. Print Printer Printer MENEE Sensorgram C None amp Current Cycle Range O AI cycles File Properties Wizard Template Wizard Results Include event log for cycles OK File Properties Wizard Template Wizard Results Sensorgram None Current Cycle Range All cycles Include event log for cycles Biqcore X100 24 3 3 3 1 Create Assay Workflow CD Kinetics Affinity Pries of recommended Aena Works Ligand details Ligand name My ligand is biomolecule with a tag an antibo another rotein poenis something else Kinetics Affinity Crea
115. 2 1 1 2 1 3 Hep TRAINING Biacore X100 Contr I File Edit View Commands Run Tc mj de E Cycle 1 Lf 24249 a ME Injection command amp l Vialwell position i Contact time s Vial well position Vial well position 1 e Contact time 3 Minimum required volume in vial well for this injection 50 ul Contact time 0 OK Biqcore X100 2 17 fig 2 1 Ex AL 5 10 30 l min Detection1 2
116. Injection Sequence Injections in analysis cycle Flow Cell 1 Flow Cell 2 RA 2 Next gt Assay Conditions Sample System Preparation Pnme Prime before run Conditioning Prime before run Prime Next gt Xe Regeneration Scouting Injection Parameters Sample Name n Concentration 20 pg ml Contact time len s Injection Parameters Sample Find Sample Conditions Find Sample Conditions Enter known values 2 I v 2L C ATE AS S Next gt BiacoreeX100 128 5 Regeneration Scouting Experimental Parameters Regeneration parameters Stabilization period 0 s Experimental design Number of conditions Number of cycles for each condition v Settings 2 v Lock C Solutions Contact times Condition Regeneration solution Contact time s 1 10mM Gly HCI pH3 0 2 10mM Gly HCI pH2 0 Stabilization period Experimental design Lock Number of conditions 30 TE Experimental Parameters
117. 105 Sample 105 Sample 105 Sample 105 Sample 105 Sample 465 Buffer Full Deionized water Water C gk we mes Rack Positions Next gt Start Biacore X 100 AN Changes detected in wizard template Help Dave As Dont Save Cancel Sqve As B amp D Don t Save l Sqave Ctrl Break 3 2 30 l Standby Biacore X100 Evaluation Software
118. 69 Custom hssay Wizard Sample Table Cycle Purpose Cycle type 1 Conc 1 Conc 2 Conc 3 Conc 4 Conc 5 nM nM nM nM nM Buffer 5 Sample 5 Sample 5 Sample Sample Table Concentration unit Cycle Purpose Sample DF 0 1 Startup Undefined Add Row Conc 1nM 1 uM 5 5 2 4 12 60 300 1500 nM D 2 Next gt Biqcore X100 70 3 Custom Assay Wizard Rack Positions E Tre samples gia 915 Sample Sample 0
119. 7224HF amp amp s3 1 2 n 2 1 Kp Ko BO 1 10 10 5 0 2 1nM 1uM MAT 5 5 Ko 10 0
120. ProteinA m IgG Ligand Attachment Overview Type of assay Direct binding analyte Selected chip CM5 T Ligand name ProteinA ligand Sensor Surface Preparation Find Immobilization pH iz Reto foo iz Reto foo Overview Enter known values known values Find Sample Conditions cm Find Regeneration Conditions LA Run tod on LA Run tod on to find out Enter known values known values BN Online COM1 Temperature 25 00 9C Sensor chip CM5 Running standby remaining time 4 0 days 5 ProteinA 16 2 Paint I first TET E s Overview V8 TRAINING Biacore Results reference C 3 33 Edit Assay Workflow Results reference Immobilization buffer 10 mM Acetate pH 5 re 5 11 2007 10 14 37 AM All results Results reference O Find Immobilization pH 2 Find Immobilization pH All results Biacore X100
121. 23 H0 another tag ii e an antibody My antibody is d mouse antibody Mouse Antibody Capture Kit BR 1008 38 Q human antibody Human Antibody Capture Kit BR 1008 39 another antibody ud e another protein Immobilize 1 2 1 2 BiacoreeX100 26 3
122. 500E 9 0 4129 7 500E 9 0 1520 1 500E 8 0 5975 3 000E 8 0 1167 Finish Add Fit Model Fit P Kinetics Affinity Fit Kinetics Create Add Fit Cyrf nt Fits Model Heterogeneous Ligand i PR E Description Coe Ce Report Residuals Parameters ka1 1 Ms kd1 1 s KD1 M ka2 1 Ms kd2 1 s KD2 M Rmax1 RU Rmax2 RU Conc M 8 868E 5 0 001174 1 324E 9 1 023E 6 X Q 003565 3 485E 9 1 766E 9 1 000E 9 3 500E 9 7 500E 9 1 500E 8 3 000E 8 STU ISgd Ncoiss amp id REELT Current Fits v 2223 S Current Fits Finish Biqcore X100 82 3 LA Kinetics Affinity Fit Kinetics Create Add Fit Current Fits Model 1 1 Binding EBBBEICY Description Delete ka 1 Ms kd 1 s KD M Rmax RU Conc M c
123. 6 139 6 2 1 Desorb 1 1 25 I 20 Maintenance Tools Desorb Stqrt l Desorb i UNI A A E IF 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 Next gt 1000 BIAdesorb solution2 Full Deionized water BlAdesorb solution 1 BIAdesorb solution 2 1 5m 1 000 ul 15mm 4 ml H2O Start Desorb
124. 74 87 88 Keuc A SUBIRE t DTP RERUE 62 L H eap 155 Load Samples SR ue IRI Pos HL I ud M MI D EUM M E 10 14 18 48 118 M ee 13 14 44 114 N NS i i Tm Il IV V 32 35 38 106 109 O oracle PODS Ri 145 146 147 162 Other options ii Ni i i i 41 44 88 111 114 OAE BYT V NNN 27 33 36 39 56 61 88 101 107 110 126 131 P GSSWUOE 159 160 161 PEGHBEGNIGBSSONMIO DI fl 159 162 PERIBUNT 8 54 57 66 89 124 127 137 142 145 PARI UU Tc ARN 23 Q Quah CODLTO i m etit er eiiim erri mH HEC EHE ite iU emu p ET 76 6 3 4 R Rack Ejected RRR A 10 ROCK IMINO GO i hl Gi 10 FOCK IOCRE O SR kk 10 14 ROCK POSION S maras A UL CU ID d LIE 29 34 37 54 59 64 70 91 103 108 124 129 133 Relerence GC GG if 21 Pelerence ES iS kk 21 50 120 Relerence SUBDEIFOUCHOD oii i i iii iii 13 44 66 114 Regeneration Command sie sic o eni C t i 50 120 REIRES o RENI eem 20 150 iz eje muorer ER Po 21 68 152 ise pM o X Residudis PE OAE AA EEA SA DESA A ESEA A MA AM 0h 77 78 79 RGS EE 157 Results TOTEN Gncios ed n ati LL A AL Li LE M Lt IA IE LAE 27 53 56 39 101 107 110 EAEE T APAE PEER EN EE E E E EN N N AEREE EN A NAE LE ATE 5 6 RNN AS IOI E IN EAS ISS R EAI ES M M i c E CMS 77 79
125. 82 RUN Sn NR Ill X 77 80 82 S OO 154 SE oo NS E 79 SeNSORE ND i i i i Nii i i 5 6 SeINSOIRGNIRIMGNMNRGINOINGG oo b oi n Id dab eu UE QUON e ade 136 137 144 Sensor oran WITIOOW ii 151 152 155 154 156 157 Set Temperat US D P 1 9 Biacore9X100 Single cycle KIDeticS eiii ai ERU HUE ME IE M 66 Specify contact tITIe oe odas e oda iamen li 1 41 111 SOD mc 20 95 96 150 Standard err Or TO D ARD UN 79 Standby 6 8 17 22 30 35 38 42 46 52 56 60 64 70 92 93 104 109 112 116 122 126 130 134 137 139 141 144 Stop DataDgseis oO bana ERU EI I Um ME 145 146 147 SOP RUP oed Eid co Ha i c ER x TNT E GE E 22 30 52 92 104 122 T temperature iiia NENA ERNIO M 1 9 Two state Reaction conformation change ssesessessessessesesssessesesseosessesseseosessessenseseoseosessesseseessensosesseseoseosesseseeseoseosessese 75 4 e e el TT 6 137 144 145 INTO MR T NEIN P O i 159 US 14 159 161 164 UV So RR 79 V dol VA VI i Meyeri dol NETT ORE 16 47 117 W WS ci er 111 WOk dE RR TEE TET 150 151 WORI ON ERROR E 24 26 98 100 PA I II VII VIII IX X XI 77 79 80 RR uam E 47 72 94 117 VOU Meo il IX X XI ROO DIRE
126. Acetate pH 5 5 10 mM Acetate pH 5 N 10 mh Acetate pH24 5 10 mM Acetate pH 4 D5 HELE pH Save Biqcore X100 5 107 admin Biacore X100 Control Software Fie view Run Tools Help Service3 GM Fe le 7 amp Assay Workflow ProteinA mlgG Assay Overview Type of assay Direct binding A analyte Selected chip CM5 Y Ligand name Protein amp ligand Sensor Surface Preparation Overview Find Immobilization pH Immobilization buffer 10 mM Acetate pH 5 D aeree Immobilize verview Find S ample Conditions ETE Find Regeneration Conditions RunBinding Analysis Assay Ce Online COM1 Temperature 25 00 9C Sensor chip CMS Running standby remaining time 7 0 days Edit Assay Workflow Results reference a Immobilization pH Scouting 10 27 2008 11 14 12 AM All results Results reference Find Immobilization pH Overview Results reference C Find Immobilization pH C
127. Chip type C Maintenance Dock Chip Tools Prime Biacore X100 Evaluation Software Prime Undock BiacoreX100 control software Exit Start AlIl Programs Oracle Database 10g Express Edition Stop Database HP Solution Center amp Install Altiris Local Recovery Microsoft Update SetProgram Access and Defaults Administr HP ProtectTools Security Manager Internet Explorer Ww MSN UA Outlook Express eo Remote Assistance B Roxio Easy Media Creator 7 Basic Edition Biacore X100 146 7 7 1 Stop Database Gm o TD Administrator Stop Database All Programs Oracle cy My Documents Stop Database EO ste My Recent Documents Database 10g Express Edition Stop Database Inte met Ej ree E Outlook E Express e My Music Keg tem Control Favorites amp n d E si E Qu Biacore X100 Evaluation 3 So 9 P Control Panel Ls
128. DB Amine Jl e S Am for immobilized level Ligand solution ProtenA in 10 mM Acetate pH 5 O Specify contact time Targetlevel RU Wash solution 50 mM NaOH ij 50mM NaOH 1 2 7 Next gt Immobilization Rack Positions 70 50 mM NaOH Immobilization 126 Ethanolamine Immobilization 85 EDC Mix Fc 2 85 NHS Mix Fc 2 Empty EDC NHS min capacity 130pl Mix Fc 2 Full Deionized water Water Rack Positions BiacoreeX100 38 3 Ligand 175 pl 11 mm 50mM NaOH 70 ul 11 mm Ethanolamine 126 ul 11 mm EDC 85 pl 11 mm NHS 85 pl 11 mm NHS EDC 11 mm
129. 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 100E 7 0 1585 3 500E 9 0 4129 7 500E 9 0 1520 1 500E 8 0 5975 3 000E 8 0 1167 Rmax RI Add Fit Parameters Parameter Settings Fit Initial value Fit global 185 Default Fit global le 3 Default Fit global YMax Default Fit global leg Default Fit local YMaxj5 Default Rmax Rmax Fit Fit local 0 RI RI Fit Constant Initial value 0 Do Parameter Setting Ok
130. Ligand name Protein Edit Assay Workflow Sensor Surface Preparation Overview Results reference Find Immobilization pH Immobilize verview Results reference Find S ample Conditions ZA Bun to find out Find Regeneration Conditions Army Sm RunBinding Analysis Assay I Ce Online COM1 Current temp 24 92 9C Sensor chip CMS Running standby remaining time 7 0 days Run to find out Run Overview Results reference Enter known values Overview Biacore X100 102 5 5 2 pH pH C 2 VI Sensor Surface Preparation
131. Rack Positions Biqcore X100 5 109 Ethanolamine 126 ul 11 mm Ligand 75 ul 11 mm EDC 85 ul 11 mm NHS 85 ul 11 mm NHS EDC 11 mm EDC NHS Next gt fete Start Sqve Ctrl Break 5 2 104 admin Biacnre X100 Control Software Immobilization 10 27 2008 11 56 13 AM Ez File Edit View Run Tools Help Service3 s W eN F zy Cycle 1 v Curve Sensorgram Fc 2 ProteinA Immobilized T Z o a w 15000 500 2000 Fe Time Window AbsResp SD Slope RelResp Bas
132. View Show Only Current Curve View Show All Curves View Show Curves of Same Type Biqcore X100 GARS 10 30 ul min ur Detection1 2 P Nr E ko Support Navigator 3 leq Standby REZAD HM lb
133. autormmaticaity when you open data for evaluation Click the bution to create an 3dG0onal entorgram window Use me 5ensorgram window to examine he Sensorgrams for interaction characterisscs and data quality Exciude any sensorgrams that are ear amp sturbed or deviate irom the expected behavior Cydes Mal you exdude at this stage will not appear in subsequent evaluations Using the ensororam window BiacoreeX100 3 65 3 6 TRAINING Biacore X100 Control Software Create Assay Workflow LJ Kinetics Affinity Binding Analysis a S Other Options gt Wizards Open New Wizard Template t Surface Preparation l4 Immobilization pH Scouting bz Immobilization L7 Assay Development l4 Assay Conditions Capture l4 Assay Conditions Sample z Regeneration Scouting Concentration Analysis Kinetics Mfinity Binding Analysis idi Custom Assay Wizard Mame Modified Created By Type Workflow L A c
134. dud td Mud VII Current FIS dais decore dae E Ha edad Op d dE RENI IRI MED da d a RN EE DERE 81 Customidetliods xv vesc ras e DECR Le i DN D SNO E D I MD i 42 112 CUSIONM REDOrGROINTS RU SLUT EE E 96 Cvele PUDO SOn noiae M M 69 D DEE OD a EEN O TUER 139 DesSorB dnd ONZE A iN i 140 142 DNUS US Ni fl V VII OGK NE ui Du DUM cM MM D E 5 137 145 E EDC e M IV 35 38 109 EndimanuUd ON T E M E 52 122 Ev dludtiorEexDIOFG ed nd t OH dt eic o Ota de m 150 151 Evaluation EXPO el aue e DUE CO de eam kk ue aU Ut dE 78 87 97 135 150 157 ERDOM T 157 F OW DO ERE NN 13 44 52 67 68 114 122 FOW TOTO iapa AN i Ni AA EM fedt 13 G GANONG Surroi i m O A I 155 H Heterogeneous ANGIVE a RN RS 75 Heterogeneous B e o ajo o SNR EIER 75 Imimobihzation DHS COUINO se edo i pb ei i oen i en HE VI 32 106 MODI acea nei ad aou db add aate ciim da tmn 25 26 34 36 37 39 99 100 108 110 Mecon COMANA seen vederlo diu ne eto buds dtu dette en Mrd tium ee one LUE 16 18 19 47 48 117 118 BiacoreeX100 K da RR RR Hl IX X 77 78 80 4 Tr c RM Hl IX X 77 78 80 KDou d i ics E hcc xL x Ui mu Hl VII IX X XI Keyword TAPIE e 72 94 IHE S ouod E A EINO E AAE I E IM uM IA MUR 24 66
135. pH3 0 135 10mM Gly HCl pH2 0 Full Deionized water Rack Positions Next gt Start Save amp 2Uv IFD AE RASLE LENDSI Ctrl Break 5 2 104 Fc Tme Wow Rep SO LRSD Sepe FRmwReop Base hkh 5 269764 03 0019 015 00 Yes 2 570 2 239 26713 04 009 2024 3349 No Temperature 25 00 lt Running regeneration scouting 1 Biacore X100 130 5 Regeneration Scouting Results Display 1stcycle Conditions BP Sample Response Baseline Result Standby Trend Chart Response Baseline Display Conditions Sensorgrqms Display Cycles Biqcore X100
136. 0296 Chip IFC type IFC5 Close Biqcore X100 1 7 1 6 Biacore X100 Sensor Chip CM5 certified Sensor Chip CM5 research grade Sensor Chip CM5 research 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 3 BR 1000 12 3 BR 1000 14 1 BR 1003 99 3 BR 1005 39 3 BR 1005 41 3 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 BR 1000 34 1 BR 1004 07 3 BR 1005 42 Biqcore X100
137. 06 1493 0 No Immobilized Ligand Protein Method Amine j Procedure TimeandFlow 25 00 C Sensor chip CMS Running standby remaining time 6 9 days Standby RESNA Immobilization Results Biqcore X100 igand FT Response flow cell Procedure Method ound RU Final RU Time and Flow Amine Protein i n 1493 0 Response Bound Response Final RU tE 3 4 Response Bound Final 2 Bound Final NHS EDC Final Bound NHS
138. 100 1 5 1 2 1 2 1 Dock Chip Dock Chip 9 Mew chip Q Reuse chip Mew chip Cip type Chip id 07051041957 emeeeeee New chip Q Reuse chip E 1 4 Chip type Chip id AFE DEELT Chip lot no sensor chip aA d dd D Dock Chip Dock Chip t Biacore X100 Control Software Biqcore X100 6 1 Dock D I
139. 20 w ug ml Regeneration is needed Save Settings Save Ligand Attachment Overview Type el mag Lireer basm ETT Selected chip CHeereiew megk butter 10 mM Eee pH 5 mhr as de Grena lrerebihr ed m de 2 451 3 RU Chap id 2051 120318 F ind Sampln Crastina m Bur T E prise Mi reo mem Vind Regeneration Conditor tal Runde bed out Eii tian values Run KineticsiMleity Aaaar e Ter aerature 2600 x enger dip CHE Running rimdbsy remaining bme 4 2 days Assay Find Sample Conditions Run to find out Overview BiacoreeX100 3 57 Find Regeneration Conditions Run to find out Enter known vaqlues Regeneration Scouting Injection Sequence Flow cell Injections in analysis cycle Flow Cell 1 Flow Cell 2 R
140. 3 Help Next gt Biqcore X100 5 133 Binding Analysis Injection Parameters First sample Contact time s Dissociation time s First regeneration Solution 1 mM Gly HCI pH1 5 Contact time s Stabilization period s Injection Parameters Next gt PE Binding Analysis Samples Sample table 05 inmiM ab HCl aH Full Delanized wer Rack Positions Next gt BiacoreeX100 134 5 Start Save Ctrl Break 5 2 104 Standby
141. 36 2643 192 N A 13 Cycle 2 2 4E 09 30 11695736 2 114536 14 1 2E 08 9 153725 15 6E 08 58 73426 16 3E 07 219 0913 17 1 5E 06 274 0177 18 19 20 BiacoreeX100 158 8 8 7 File Save ASs Save in folder Folder Lr nakayama a e Users Name Modified Crea Type S E Training Geval 5 23 2007 2 40 09PM_ Training Evaluation B Group B3 murain E Nakano id ly Norio Group E Shemy Yoshimi E Software exercises E system check E Yoshitani administrator amp Instrument Service ss Cere Name Sqve Biqcore X100 9 159 9 9 1 Administrators Users 1 Administrator Username admin Password administrator User Administrators
142. 4 es E E7tvk amp Huco k amp ik tnU RERE CRI S S ER de Biqcore X100 8 149 8
143. 45 ProteinA Sample 10 mM Acetate 4 135 50mM NaOH Regeneration Full Deionized water Water Rack Positions Next gt Start Save 3 2 30 Biacore X100 30 3 E 3 2 Run gt Stop Run Biacore X100 A This will stop the run mp Lsewwl core Stop Run Run Stopped Finishing current cycle please wait Abort cycle by Ctrl Breakl Standby Ctrl Break Biacore X100 2 You have aborted the run Do you want to wash the system Recommended CeCe Yes
144. A Biacore Biacore surface plasmon resonance SPR Biacore B ORENT Biqcore X100 Il
145. A 2 Next gt Assay Conditions Sample System Preparation Prime Prime before run Conditioning Prime before run Prime Next gt Concentration 20 pg ml Injection Parameters Find Sample Conditions Find Sample Conditions Enter known vaqlues Next gt Biqcore X100 58 3 Regeneration Scouting Experimental Parameters Regeneration parameters Stabilization period 0 s Experimental design Number of conditions Number of cycles for each condition v Settings 2 v Lock C Solutions Contact times Condition Regeneration solution Contact time s 1 10mM Gly HCI pH3 0 2 10mM Gly HCI pH2 0 Stabilization period Experimental design Lock Number of conditions 30 TE Experimental Parameters Regeneration parameters
146. Biqcore X100 3 35 Ethanolamine 126 ul 11 mm Ligand 75 ul 11 mm EDC 85 ul 11 mm NHS 85 ul 11 mm NHS EDC 11 mm EDC NHS Next gt Start Saqve 2U v 2S S Ee sERISIEUEUMSSla Ctrl Break 3 2 30 admin Biacore X100 Control Software Immobilization 10 27 2008 11 56 13 AM Ez File Edit View Run Tools Help Service3 s W eN F zy Cycle 1 v Curve Sensorgram Fc 2 ProteinA Immobilized T Z o a w 15000 500 2000 Fe Time Window AbsResp SD Slope RelResp Baseline Id Keywords in cycle 1 Value 204634 0 05 0 01 0 0 Yes Baseline Chip CM5 5 206225 0 16 0 06 159 1 No EDC NHS ContactT ime 420 5 218185 058 0 31 14551 No Ligan 5 5 gand FlowRate 219564 011 0
147. E OANA HE SP e c6REOXS e5X33J2 QU e o Pe OFF
148. E 9 641ET7 1 000E 9 30 00 S 100E 7 0 1585 3 500E 9 0 4129 S DE 9 0 1520 1 5 E 8 0 5975 3 DE 8 0 1167 ka 1 Ms ka 1 s Ko M Rmax RU RI RU bulk effect Chi2 RU2 Residuqls G Parameters 3 16 81 Finish Evaluation Explorer x Remove Ei 7 Sensorgram J All sensorgrams Plot a Baseline Report Point Table ri Report Paint T able q gt Kinetics Affinity AJ z488 AJ Kinetics Affinity Biacore X100 3 79 3 15
149. GE Healthcare Biacore Biacore X100 Plus Package Instrument Handbook GE imagination at work 71 3190 31 kk A Biacore ee 7 NEED C KA CSE cc Gc II ChP anaa ana IV C 2 pH R Lettere tenete tette ttt tt ttt ttt totis VI D ee VII D 1 I kd3 pilu E A52 OB VII D 2 Lssseeenntententnntnntentnntettnettenttentnes IX D 3 Ek XII PELO EIU S 1 1 1 kk 1 iii EROTS ET aen a R 1 1 1 2 ouusentennttnnennttnnttnnttnnnttnnttnnnes 1 1 1 3 ee 2 1 1 4 kk 3 1 2 4 5 1 2 1 p 9 39 7 BAN oA A i 5 1 2 2 27 7 RIC d 3EIGBHE io etetrettenttnettrntteetentttnntnnctis 8 1 2 3 DIM E Ehou secte uM EAM a M A AMA AU I 9 1 2 4 89 JJ boo tow 62V DJODRUOTCB o cito t cera ER ties 10 1 3 aiANAl i ll i 11 2 i 13 Biacore X100 2 1 ae 13 2 1a
150. Hacnrn X100 Contrel Tada tal ur TAA RU ed PMA Ee pde Wee Commands Run Took jip Savie Eis 1 re Geriona Fes eremralmnm 7M 3 pg 415 RU N Reference Line 344 0 s 41 5 AU 200 an mmigw Abe SOO LRSD Siye ReRep Haw 5 hik dar m tt d mem 1 5 KAS NF QI Me Hl Me 1 1 Iw 553 MI JA MDE Mo Hiahmr 1 amp Biqcore X100 52 3 New Cycle ic Commands gt New Cycle Hew Cycle FI DIA Ok F Flow rate pl rin FI th ow pa Help E Flow path 1 Q Ed Flow path z Reference subtractian Flow path 1 2 2 v Flow path OK End manual run dp Run gt Stop Run Biacnre X100 v This will end the manual run when the last queued command is completed OK
151. M5 CM CM 0 5 pH pH Immobilization pH Scouting Immobilization pH Scouting AIRO pH pH
152. NaOH Regeneration Full Deionized water Water Rack Positions Next gt Start Save 5 2 104 BiacoreeX100 104 5 E 5 2 Run gt Stop Run Biacore X100 AN This will stop the run Stop Run Run Stopped Finishing current cycle please wait Abort cycle by Ctrl Break Standby ctl Break Biacore X 100 3 You have aborted the run Do you want to wash the system Recommended Yes
153. R Regeneraqtion Command Regeneration Wal position lg UK Contact time 6 Minimum required volume in vial for thie injection Z0 pd D 1 admin Biacore X100 Control Software Manual Run 10 27 2008 2 14 43 PM File Edit view Commands Run Tools Help Service3 m B Cycle 1 Curve Sensorgram Fc 2 RR hebp PP iO L inding 1 f stability 1 Response f baseline 1 0 100 200 300 400 600 Fc Time Window AbsResp SD LRSD Slope RelResp Baseline Id Keywords in cycle 1 Value 2 1750 5 219736 nd 007 0O01 0 0 Yes baseline 1 2 30 0 5 2438976 2183 015 11 67 24240 No binding 1 2 350 5 244582 663 013 3 54 24855 No stability 1 Flow 30 Flow path 1 2 Online COM1 Temperature 25 00 C Running manual run View Reference Line 3 l Biqcore X100 5 121 XM To P AF bw 1 v a E LLEN Wu Reference Line 156 0 s 0 0 AU Fc Tme Window Abfle p SD 5 Slpe Refleap Baseline Id 5 ANNE o0 00 Yer baseline 1 5 3976 2183 24240 No bris 1 5 245382 66 24856 No stab
154. Response v Bar Chart amp Ctrl oe smpswaws Buffer Buffer Buffer sample1 sample2 sample3 sample4 sample5 sample6 sample7 sample8 sample8 1 2 3 4 5 6 7 8 9 10 11 12 l Evaluation Explorer c Sensorgram hm rd All sensorgrams 5 Plot 4 Binding level Sample gi Binding stability Sample 4 Binding to reference Sample E gt Report Point Table mm Repert Peint Table C Binding Analysis M a Binding Analysis Biacore X100 136 6 6 Tools gt More Tools Maintenance Tools
155. X100 Control Software Manual Run 10 27 2008 2 14 43 PM File Edit view Commands Run Tools Help Service3 m B Cycle 1 Curve Sensorgram Fc 2 RR hebp B ok ee iO L inding 1 f stability 1 Response f baseline 1 0 100 200 300 400 600 Fc Time Window AbsResp SD LRSD Slope RelResp Baseline Id Keywords in cycle 1 Value 2 1750 5 219736 nd 007 0O01 0 0 Yes baseline 1 2 30 0 5 2438976 2183 015 11 67 24240 No binding 1 2 350 5 244582 663 013 3 54 24855 No stability 1 Flow 30 Flow path 1 2 Online COM1 Temperature 25 00 C Running manual run View Reference Line 3 l Biacore X100 3 51 dmin lscnrm X100 Control Software Manual Run 10 77 72000 7 14 43 PH Gpe Ea ye Commands Eun Toos Hep Sene g 70 Frp gt pe 1 Ope Seniogram F c 2 Reference Line 156 0 s 0 0 AU mm Fc Tme Window Ab esp SD LASD Skpe Refleap Baneine Id EE ncyde Value 5 nwi6 0y 007 00 Yet baseime 1 5 3976 2183 015 24240 No bris 1 5 24532 663 013 24056 No stablity 1 U2TPUZA24vof i 4h9702Uv2 amp HF2v7Z7cpzo2d4j9huAmmiI ct8 ct View Baseline 0 l ES admin i
156. c 1 9 583mB02 Yy LERZ 272 KOF User account L Account is locked Account is disabled OK Quilt BiacoreeX100 162 9 9 2 Biacore X100 Control Y Biacore X100 Evaluation Oracle Database 10g Express Edition 23 Administrators 2GB Biacore X100 Control Software Tools Preferences Database Backup Preferences Schedule C Monday Tucsday Wednesdap Thursday Enida Saturday F Sunday Stat time ng Backup rw Backup l
157. cei ee Lee C AME E ee eie AERE EE 6 i ondas 3 5 7 88 pus Ais LD A RR ETT 5 e M IIl 88 A e AE EN IV 10 16 18 35 38 47 48 109 117 118 139 143 IESE dica ERR IER ET NR ERR E A EE Il VII IX X 75 79 81 zz DACIA TNT RT X zz Ecl CE NRI uu VI VII arci MMC 79 KS PAV TA A aaa a 77 79 80 DAUT IIe T O naa E O EAR A RO 79 N 612 X 84 MAAA E BEES om by 142 Biacore X100 c EME RERO 44 114 ER E T E IX ch V VIII acc e 0 2 159 ck eee VII 77 78 87 10 14 17 46 116 PI IIP EEE a o TE REO ER DDR 8 ee 8 I aik II HI IV V VI VII VIII IX 79 80 82 DE aD d pone RE V VI 28 32 102 106 939 o CIRCO DH OS US ciues etos de oo eb tdi sU Lee EA De Ae CU EEDE VI 3 NT Agr aer aed REDI RT PETRI VII 77 Xl 35 38 109 oo IX 4 11 12 24 25 26 28 33 34 36 37 39 41 44 53 56 61 62 63 65 88 98 99 100 102 107 108 110 111 114 125 126 151 132 BiacoreeX100
158. cm ui Notepad Q irem Access and vicc TFAU Cd C02 TT WW Help and Support Search 7 Run HP Support All Programs D Log Off Turn Off Computer pos Oracle Database The OracleServiceXE service was stopped successfully DOS amp Stop Database RAI aclexe app orac le pr nu 1 guETKHBTNPnEt stop Orac EET UTR DE E The racle ervice E service is stopping E The racle eryvice E service was stopped successfully Ci oraclexer app rorac lesnproductsi z EE BI Windows Shutdown d 2 Biacore X100 PC Oracle Database 7228829 g 2 Biqcore X100 7 147 E 7 2 Oracle Database Stop Database Oracle Database
159. e Copy of Amine Command Solution Contact Time s Flow Rate l min P PRE CONC Specified in Immobilization Setup 6 EDCNHSINJECT EDC NHS 50 50 A wasH Ethanolamine L LIGANDINXECT Specified in Immobilization Setup 4 maEcT Ethanolamine Method Method name Edit Custom Methods Copy of Amine 0 OE of Amine Ke Surface thiol Method name Copy of Amine Command Solution Contact Time s Flow Rate l min 79 PRE CONC Specified in Immobilization Setup 5 EDCNHSINJECT EDC NHS 50 50 A wash Ethanolamine D LIGANDINJECT Specified in Immobilization Setup INJECT Ethanolamine OK Set up Method Biqcore X100 44 3 3 4
160. e rack and click OK Time until automatic standby 4 52 Rack ea OK 13 rqck Iocked Tools Rack Illumination 1 9 Position HO 15 _Position 1 position 1 position 15 X position 15 2 Milig position H20 1 Rack position 1 a45 0 e caps type a BR 1004 11 1 5ml Plastic Vials 15mm Plastic Vials BR 1002 87 BR 1006 54 Biqcore X100 1 11 1 3 Biacore X100 3
161. eline Id Keywords in cycle 1 Value 204634 0 0 5 0 01 0 0 Yes Baseline Chip CM5 5 205225 0 16 14 0 05 1591 No EDC NHS ContactTime 420 5 219185 0 58 i 0 31 1455 1 No Ligand FlowRate 5 5 219554 011 0 06 X 14930 No Immobilized Ligand Protein Method Amine j Procedure TimeandFlow 25 00 C Sensor chip CMS Running standby remaining time 6 9 days Standby RESNA Immobilization Results Biqcore X100 110 5 Immobilization Results Chip CM5 igand FT Response flow cell Procedure Method ound RU Final RU Time and Flow Amine Protein i n 1493 0 Response Bound Response Final RU tE 5 4 Response Bound Final 2 Bound Final NHS EDC
162. ffinity Injection Parameters Sample Contact time i20 s Dissociation time 2 3 First regeneration Cantact time 30 s Stabilizationperiod s Help Injection Parameters Next gt Kinetics Affinity samples Samples mouse IgG mouse Ig mouse IgG mouse Ig mouse IgG mouse Ig mouse IgG mouse IgG 150000 150000 150000 150000 150000 150000 150000 150000 Concentration Concentration 8 33E 3 16 7E 3 333E 3 66 7E 3 0 133 3323E 3 Sample Sample id Concentration 2 5 2 1 2 D 2 IX Samples MW Dqa Da Biacore X100 64 3 V Kine
163. heroptions Wizards Immobilization New Immobilization Setup gt Chiptype SLE v How cell 1 O immobitize fow cett 1 Flow cell 2 O z immobitize fow cell 2 Hp Csom meroa l Prime before run Method Capturing molecule ligand solution Target level RU Wash solution Method Capturing molecule ligand solution Target level RU Wash solution Chip type Method Aim for immobilized level Wash solution Specify contact time OBlank immobilization Next amp 2 v 2d S Surface thiol Target level 50mM NaOH Biacore X100 42 3
164. i 4 C 40 Cos88B CagiEL C OK set Temperature Status bar temperature 277 B TS amp 2 gd D temperature lt 1 9 Biacore X100 10 1 1 2 4 Load Samples j rack locked d rack locked Load Samples Hack Ejected Insert th
165. ize Step 4 Wipe the pump inlet tubes with a moist tissue Place water on the left hand tray and insert the pump inlet tubes ABI DLL L2Z Q amp b amp FRAT7UvkXcai 225 KRKY kg Start MERA 13 l 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 performing a run 4 Standby Stqndby 3 4 Close BiacoreeX100 142 6 6 3 Desorb and Sanitize 35 BIAtest solution e HBS EP 10X BR 1008 26 Sensor chip CM5 HBS EP CM5 D
166. le Name mouse lqG Contact time 60 s Dissociation time 130 s Max concentration 20 v ug ml v Regeneration is needed Save Settings Save ES TRAINIMG a Dincore X100 Control Software Pie View Tenis Hein zi t C Assay Workflow ProteinA m IgG I igand Attacnmanli Cerne Type el tity anale elected chup ligari name i gm La TPehirahrm hoffer 20 mM Fri pH Method m lc z arae bmmobikred in de z 451 3 ARU Chip x amp 70511 0223 et tt chip CS Ennis standby remaining be 40 dins EE TANING Emmie neinn sump 7 Er TAMK 0 Bamwr i Assay Find Sample Conditions Run to find out Overview Biqcore X100 5 127 Find Regeneration Conditions Run to find out Enter known vaqlues Regeneration Scouting
167. lity 1 View Baseline 0 l ES admin i Hacnrn X100 Contrel Tada tal ur TAA RU ed PMA Ee pde Wee Commands Run Took jip Savie Eis 1 re Geriona Fes Reference Line 344 0 s 41 5 AU fe Time Wride Abe SO LASO Sigs i 3 hie a n i 5 KAS NF QI Me Hl L 5 Iwan 553 BES y Biacore X100 122 5 Flow F Flow rate Flow path Help E Flow path 1 Q Ed Flow path z Reference subtractian Flow path 1 2 2 v Flow path OK End manual run dp Run gt Stop Run Biacnre X100 v This will end the manual run when the last queued command is completed OK standby
168. low path 1 2 Online COM1 Temperature 25 00 9C 1 2 2 1 1 View Show Only Current Curve View Show All Curves View Show Curves of Same Type BiacoreeX100 5 Ne E YE JAER Detection1 2 5 10 30 ul min Support Navigator
169. n Analysis 4 24 2007 7 161 Immobilization Conc 4 24 2007 7 16 E Immobilization pH Scouting 5 9 2007 10 17 39 AM 5 9 2007 10 32 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 26 Immobilization 5 9 2007 11 28 28 AM 5 9 2007 12 13 Immobilization 4 25 2007 11 04 44 AM 4 25 2007 11 22 Name Manual Run 5 10 2007 1 30 44 PM 9 Save Biqcore X100 ES TRAINING Biacore X100 Control Software Manual Run 5 10 2007 1 30 44 PM EzjFie Edit View Commands Run Tools Help 2 15 inj OLJA a M 0 5 10 15 Fc Tie Window AbsResp SD LRSD Slope RelResp Baseline ud 20 C Lock scale 25 30 Time R Keywords in cyde 1 Value F Flow 10 Flow path 1 online COM1 Temperature 25 00 oC Running manual run 0 V8 TRAINING Biacore i 7 Paint Biqcore X100 16 2
170. ocation Backup folder CFrogram FilssVBiacore Biacore 100 Dalabassh C Enable second copy Folder Humber of saved backups 7 5 Aesioe Avalable backups for restore based om backup Ioided backup crastad 4 1 2007 11 00 55 PW Bk Cancel Schedule Start time Backup location Backup folder C ProgramFiles Biacore BiacoreX100 Database Enable second copy Hga Biqcore X100 9 163 Number of saved backups 7 Cc Restore
171. ock Ls Prime Tools More Tools l 7 Desorb l Desorb and Sanitize Test Tools System Check and Pump Calibration B E Service Tools 1 Software Problem Report ad Flow System Wash 4i Supercean j Clean Needle and Needle Wash Station Calibrate Needle Replace Needle 4y Replace Peristaltic Pump Tube aM Deb Cami This procedure checks forleakage 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 EP4 Last run time N A Test Tools gt System Check and Pump Calibration Start 2 v 2d 2 System Check and Pump Calibration Ensure that you have HBS EF as running buffer the analysis temperature is 25 C and a new Sensor Chip CM5 is docked HBS EP HBS EP 10X 10 Next Biqcore X100 6 143 System Check and Pump Calibration Rack Positions Mem lt lead omaes me f sw Se BIAtest Solution 1 5 ml 123 ul 1
172. r chip CM Overview mm irabon buller 10 mid Acetate pH 5 Method in fc 2 Amme kmenolslued in ic 2 451 3 RU Chip id 070511 0933 Rynning standby remainies tme 4 0 days 2 220 2 HFk40X Assay Find Regeneration Conditions Run to find out LJ Overview Biqcore X100 62 3 3 5 Run Kinetics Affinity Assay Run Kinetics Affinity Injection Sequence Detection Chip Flow cell Chip type Injections in analysis cycle FlowCell Flowcell2 4 Injection Sequence Next gt Kinetics Affinity System Preparation x p Prime Prime before run prime Conditioning Startup w Run startup cycles D Buffer Number of cycles INI v Solution 3 Next gt BiacoreeX100 3 63 Kinetics A
173. re X100 2 1 1 1 3 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 eFRoBIe EKT 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 eFRoBIe EKT 10 HBS N 10X 50 ml 4 BR 1008 28 0 1 M HEPES 1 5 M NaCI gt 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 NaCl 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 RREZEN RERO CHS 0 22 um Biqcore X100 1 3 1 1 4 Start Programs Biacore BIACORE X100 Control Software
174. reate Assay Workflow Binding Analysis Ligand details Preview of recommended amp ssay Workflow Ligand name Protein My ligand is another protein 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 Direct binding analyte Selected chip CM5 Na Ligand attachment overview Preview of recommended Assay Workflow DRN Continue Save Assay Workflow As Save in folder Folder G nakayama Users Name Modified 3 8 Training ProteinA mlgG2 5 9 2007 2 26 05 PM i ProteinA mlgG 5 9 2007 9 58 30 AM j 070509 5 9 2007 9 27 23 AM 2 5 8 2007 2 31 33 PM i5 5 8 2007 2 29 32 PM i4 5 8 2007 2 08 54 PM ji 5 8 2007 2 08 04 PM Biacore X100 5 101 KW admin Biacore X100 Control Software File view Tools Window Help Assay Workflow ProteinA_mlqgG 77 4 Y Type of assay Direct binding Selected chip CM5
175. s another protein v Ligand attachment approach Recommended O Capture using own antibody and Sensor Chip CM5 O Immobilize ligand covalently using Sensor Chip CM5 Continue Cancel BiacoreeX100 5 99 fig 5 1 Ligand details My ligand is Ligand attachement qpproach a biomolecule with a tag My ligand is tagged with biotin Sensor Chip SA GST GST kit BR 1002 23 another tag A e an antibody My antibod
176. sorgram 300 Biqcore X100 8 157 8 6 Sensorgram window Export Curves txt Microsoft Excel File Export Result To Excel Evaluation Explorer xls Microsoft Excel xls Microsoft Excel nau B EvaluationsKinetics Affinity 2 Name Sample 3 4 Sample Sample 5 Temperat 25 6 Curve Fc 2 1 z 8 Model Bivalent Analyte 9 Description 10 11 Curve kal 1 Ms kdi 1 5 ka2 1 RUskd2 1 5 Rmax RU Conc M tc Flow ul mkt RU MS RI RU Chi RU2 U value 12 28071 45 0 018845 0 000287 0 06355 15688 84 37640
177. tar known Method in fc 2 Amme kmenolslued in ic 2 451 3 RU Chip id 070511 0933 oc ge Find Sample Conditions kd ig Puto fec c Erter roue values J Find Regenerabon Condi ons m Run 15 fird aa Erim kroan valusa j Sensor chip CM Rynning standby remainies tme 4 0 days 2 220 2 Hk40X Assay Find Regeneration Conditions Run to find out LJ Overview BiacoreeX100 132 5 5 5 Run Binding Analysis Assay Run Binding Analysis Injection Sequence Detection Flow cell Chip type Injections in analysis cycle FlowCelli Flow Cell2 ce wg Enhancement v Regeneration 1 w Injection Sequence 91 D 75 Xineti 27 220 CZSEUJSZEUE D BISIDERR GS Next gt Binding Analysis System Preparation Prime Prime before run prime Conditioning Startup Run startup cycles Solution Buffer YZ YTRER APA Number of cycles 3
178. te Assay Workflow Kinetics Affinity Choose ligand attachment approach Preview of recommended Assay Workflow Ligand details Ligand name ProteinA My ligand is Loherpan Recommended Q Capture using own antibody and Sensor Chip CM5 Q Immobilize ligand covalently using Sensor Chip CM5 Emus ca BiacoreeX100 3 25 3 1 Ligand details My ligand is Ligand attachement qpproach a biomolecule with a tag My ligand is tagged with biotin Sensor Chip SA GST GST m kit BR 1002
179. tics Affinity Rack Positions 105 mouse IgG 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 2 3 4 5 6 ri 8 H gt E EE Rack Positions Next gt l Start l Suve Ctrl Break 3 2 30 l Standby Biacore X100 Evaluation Software we TRAINING Biacore X100 Evaluation Software Kinetics Affinity 3 14 2007 2 09 23 PM Support Navigator Pie View Evsuston Toos Window Help lt lt Prnt gt hi gt Semyan jAk A Knescs Minty Brang Analysis A Concentraton Analyss What do you want to do Evaluate kinelics affnit Evaluate tinding analysis Evatuste concentration anahrsis Sensorgram window This window i5 displayed
180. urpose Pe Startup Q Apply To Selected Cycles Calibration Sample Id Cycles 4 4 Calibration Table Exclude Cycle Import Cycle Biqcore X100 Samples Ws Concentration Analysis Create Calibration Samples Sample Flot RU 2000 22 samplei 1000 i 5173 26 sampnle1 1000 1800 5172 1600 23 sample2 1108 27 sample2 1400 NENNEN 1108 1200 24 sample3 187 4 28 sample3 1000 Relative Response 800 25 sample4 29 sample4 600 400 200 1 5 2 2 5 3 3 5 4 5 5 5 5 Measured Concentration pg ml Sample Table Cycle Sample id Dil Fact T AES AE Response RU efek Calc Conc
181. y is d mouse antibody Mouse Antibody Capture Kit BR 1008 38 Q human antibody Human Antibody Capture Kit BR 1008 39 another antibody po e another protein Immobilize 1 2 HJI 1 2 Biqcore X100 100 5 Immobilize ligand covalently using Sensor Chip CM5 C

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