兔单克隆抗体了解兔单克隆抗体的精彩课件

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1、2023-2-31Rabbit Monoclonal Antibodies-Better Tools for Better Science2023-2-3 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 high

2、 homology to human antigensHigher throughputStable hybridomasMultiplexHigh humanization probabilityHigher homology to human germline sequence than mouseA in vitro positive proof-of-concept study2023-2-3Rabbits Recognize More EpitopesMouseRabbit1.Molecular-weight marker2.Fibronectin3.Protease digeste

3、d fibronectin1 2 3 1 2 35 mice and 3 rabbitsWestern AnalysisPolyseraImmunized with human fibronectin2023-2-3Supernatant 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 MAb2023-2-3A Random Collection Of RabMAbs Have Extremely High Affini

4、tiesAntigen MAbKon(1/M.sec)Koff(1/sec)Kd(M)EREPIer392.58x1052.45x10-39.5x10-9EREPIer142.03x1059.28x10-54.57x10-10EREPIer261.72x1053.37x10-52.19x10-10EREPIer382.87x1057.79x10-42.71x10-9PREPIpr421.18x1061.63x10-21.38x10-8PREPIpr333.22x1064.13x10-61.28x10-12PREPIpr884.46x1052.51x10-45.63x10-10C29EPIc29

5、027.61x1042.88x10-33.78x10-8C29EPIc29033.26x1043.12x10-69.57x10-11C29EPIc29051.36x1054.96x10-43.65x10-9C29EPIc29063.43x1044.57x10-51.33x10-9Herceptin Kd(M)=5.0 x 10-9$300MRituxan8.0 x 10-9$1.1BSynagis1.0 x 10-9$600MRemicade2.0 x 10-10$270M2023-2-3RabMAb For Better DiagnosticsMouse 1A6RabMAbHeat trea

6、teduntreated2023-2-3Rabbit MAbMouse MAbPhage MAbNon Primary AbRabMAb:More Accurate Results-Comparison of rabbit MAb,mouse MAb and phage MAb50%false positive15%false positive5%false positive2023-2-3Fibrin Degradation ProductsFibrin(Blood clot)PlasminPlasminogenEndothelia CellPAI-1tPA PropertyApplicat

7、iontPA RabMAb clone 8BindDiagnosticsclone15InhibitTherapeuticsclone22StimulateTherapeuticsclone29Strong stimulateTherapeuticsclone34BindDiagnostics PAI-1 RabMAb clone2Inhibit PAI-1/tPATherapeuticsclone7BindDiagnosticsclone10Inhibit PAI-1/tPATherapeuticsclone12Inhibit PAI-1/tPATherapeuticsclone14Bind

8、DiagnosticsOne Process:Multiple RabMAb ProductsPAI-1tPA2023-2-3PathoMAb to Identify Therapeutic MAb and Disease TargetTumor cellEmbryonic cellInflammatory tissueDegenerative tissueMembrane fractionNuclear fractiongenerates up to 10,000 MAbs from disease tissues directly YYYYYAntigensBioactivityThera

9、peutic MAbcandidatesDiseasetargetsCell-based assays2023-2-3MAbs 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 cells 2 an

10、tibodies 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 2023-2-300.040.080.120.160.2C O N TR O LC O N TR O L18A18A22C 122C 122C 222C 241H41H15E15ER abbi

11、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 Adhesion2023-2-3Primary tumor(20%expression)Lung metastasis(100%expression)RabMAb To A Novel Ion Channel Tumor Antigen-identified directly from immunization of cancer tissues2023-2-3Using Subtractive Path

12、oMAb 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 MAbs2023-2-3A Kidney Specific Antibody Generated by PathoMAbBrainLungLiverKidney2023-2-3Proof Of Principle For Hu

13、manized RabMAb-from 75%to 95%human sequenceHzdMAbRabMAb293 b b-6 transfectants293 untransfected2023-2-3Market Demand For RabMAbsSmall MoleculesPeptidesMTMProteinsMouse MAbs Unavailable-Novel Research reagents or leads for research,drug discovery and diagnostics.Mouse MAbs Available-Opportunity for b

14、etter quality follow-on antibodiesHigher AffinityDiagnosticsNovel EpitopesHigher AffinitiesTherapeuticsAntibody PairsResearch ReagentAntibody Arrays2023-2-3Improve The Probability Of Success For Drug DiscoveryRabMAbTechnologyTarget ID&ValidationProtein level,Clinical-directedtarget validationMultipl

15、ex,Companion diagnosticsNovel Epitope,High affinityTherapeutic leadsDiagnostic leads2023-2-3Proteins27%DNAs2%Small Molecules1%Peptides70%ProteinsPeptidesSmall MoleculesDNAsSuccess Rate Of RabMAb GenerationAntigensTotal AntigensAntigen-Binding mAbs Desirable mAbs Overall Success RateProteins34323191%

16、Peptides96898083%Small Molecules22150%DNAs62233%Total138125114Finished projects138On-going projects177Total projects3152023-2-3 2023-2-3Summary 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.