兔单克隆抗体-了解兔单克隆抗体的精彩课件.ppt

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1、2022-12-61Rabbit Monoclonal Antibodies-Better Tools for Better Science2022-12-6 Advantages of RabMAb Technology Over Mouse MAb TechnologyHigher quality High affinity and specificityBroad applications High success rate in meeting customers criteriaNovel Epitopes Recognition of mouse orthologs with hi

2、gh homology to human antigensHigher throughputStable hybridomasMultiplexHigh humanization probabilityHigher homology to human germline sequence than mouseA in vitro positive proof-of-concept study2022-12-6Rabbits Recognize More EpitopesMouseRabbit1.Molecular-weight marker2.Fibronectin3.Protease dige

3、sted fibronectin1 2 3 1 2 35 mice and 3 rabbitsWestern AnalysisPolyseraImmunized with human fibronectin2022-12-6Supernatant From Rabbit Hybridoma Is Highly Active In Western blotABCD(A=1:5;B=1:50;C=1:500;D=1:1000)K8Anti-Human Keratin 8 MAb2022-12-6A Random Collection Of RabMAbs Have Extremely High A

4、ffinitiesAntigen MAbKon(1/M.sec)Koff(1/sec)Kd(M)EREPIer392.58x1052.45x10-39.5x10-9EREPIer142.03x1059.28x10-54.57x10-10EREPIer261.72x1053.37x10-52.19x10-10EREPIer382.87x1057.79x10-42.71x10-9PREPIpr421.18x1061.63x10-21.38x10-8PREPIpr333.22x1064.13x10-61.28x10-12PREPIpr884.46x1052.51x10-45.63x10-10C29E

5、PIc29027.61x1042.88x10-33.78x10-8C29EPIc29033.26x1043.12x10-69.57x10-11C29EPIc29051.36x1054.96x10-43.65x10-9C29EPIc29063.43x1044.57x10-51.33x10-9Herceptin Kd(M)=5.0 x 10-9$300MRituxan8.0 x 10-9$1.1BSynagis1.0 x 10-9$600MRemicade2.0 x 10-10$270M2022-12-6RabMAb For Better DiagnosticsMouse 1A6RabMAbHea

6、t treateduntreated2022-12-6Rabbit MAbMouse MAbPhage MAbNon Primary AbRabMAb:More Accurate Results-Comparison of rabbit MAb,mouse MAb and phage MAb50%false positive15%false positive5%false positive2022-12-6Fibrin Degradation ProductsFibrin(Blood clot)PlasminPlasminogenEndothelia CellPAI-1tPA Property

7、ApplicationtPA RabMAb clone 8BindDiagnosticsclone15InhibitTherapeuticsclone22StimulateTherapeuticsclone29Strong stimulateTherapeuticsclone34BindDiagnostics PAI-1 RabMAb clone2Inhibit PAI-1/tPATherapeuticsclone7BindDiagnosticsclone10Inhibit PAI-1/tPATherapeuticsclone12Inhibit PAI-1/tPATherapeuticsclo

8、ne14BindDiagnosticsOne Process:Multiple RabMAb ProductsPAI-1tPA2022-12-6PathoMAb to Identify Therapeutic MAb and Disease TargetTumor cellEmbryonic cellInflammatory tissueDegenerative tissueMembrane fractionNuclear fractiongenerates up to 10,000 MAbs from disease tissues directly YYYYYAntigensBioacti

9、vityTherapeutic MAbcandidatesDiseasetargetsCell-based assays2022-12-6MAbs Against Mouse Mammary Tumors Raised From A Single Rabbit7000 hybridoma clones generated:157 of 1152 antibodies are useful for immunohistochemistry80 antibodies preferentially recognize tumor57 antibodies react with epithelium

10、cells 2 antibodies bind specific for necrotic areas of tumor11 antibodies react with stroma2 antibodies stain cell-cell junctions 5 antibodies detect secreted molecules10 antibodies react with myoepithelial cells 2022-12-600.040.080.120.160.2C O N TR O LC O N TR O L18A18A22C 122C 122C 222C 241H41H15

11、E15ER abbi t B l ocki ng M A bsR abbi t B l ocki ng M A bs570nm O.D.570nm O.D.Blocking MAbs for Tumor Cell Adhesion2022-12-6Primary tumor(20%expression)Lung metastasis(100%expression)RabMAb To A Novel Ion Channel Tumor Antigen-identified directly from immunization of cancer tissues2022-12-6Using Sub

12、tractive PathoMAb to Identify MAbs Specific to Disease TissuesImmunize Rabbit withLung Cancer CellsScreen Hybridoma Against Lung Cancer Cells Back-Screen Positives Against Normal Lung Cells 100 MAbs5 MAbs2022-12-6A Kidney Specific Antibody Generated by PathoMAbBrainLungLiverKidney2022-12-6Proof Of P

13、rinciple For Humanized RabMAb-from 75%to 95%human sequenceHzdMAbRabMAb293 b b-6 transfectants293 untransfected2022-12-6Market Demand For RabMAbsSmall MoleculesPeptidesMTMProteinsMouse MAbs Unavailable-Novel Research reagents or leads for research,drug discovery and diagnostics.Mouse MAbs Available-O

14、pportunity for better quality follow-on antibodiesHigher AffinityDiagnosticsNovel EpitopesHigher AffinitiesTherapeuticsAntibody PairsResearch ReagentAntibody Arrays2022-12-6Improve The Probability Of Success For Drug DiscoveryRabMAbTechnologyTarget ID&ValidationProtein level,Clinical-directedtarget

15、validationMultiplex,Companion diagnosticsNovel Epitope,High affinityTherapeutic leadsDiagnostic leads2022-12-6Proteins27%DNAs2%Small Molecules1%Peptides70%ProteinsPeptidesSmall MoleculesDNAsSuccess Rate Of RabMAb GenerationAntigensTotal AntigensAntigen-Binding mAbs Desirable mAbs Overall Success Rat

16、eProteins34323191%Peptides96898083%Small Molecules22150%DNAs62233%Total138125114Finished projects138On-going projects177Total projects3152022-12-6 2022-12-6Summary RabMAb is a superior technology platform that is widely accepted by the industry Epitomics enjoys rapid growth in customers for research,diagnostic and therapeutic fields Rabbit MAbs have high affinity;Rabbit hybridomas are stable;The success rate to obtain“desirable”MAbs is high;The throughput can be high.

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