LEC04PH - 1 Immunology 2011 Lecture 4 Antibody Structure II...

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Unformatted text preview: 1 Immunology 2011 Lecture 4 Antibody Structure II 22 September Different epitopes -- > different Abs (heterogeneity of Abs & epitopes) Principles of Ag/Ab Reactions Binding is reversible (equilibrium) Antibodies bind epitopes (=antigenic determinants) Definitions gamma-globulin electrophoretic mobility immunoglobulin (Ig) chain & domain structure antibody (Ab) binds Ag Measuring Antibody/Antigen Reactions Binding is invisible: how to detect & quantitate ? 1) Precipitation: Ouchterlony 2) Solid-state binding, labelled antibody: RIA/ELISA 4) Consumption of complement: Complement Fixation (read-out is hemolysis) 3) Liquid-phase binding: Equilibrium Dialysis Measuring Antibody/Antigen Reactions Binding is invisible: how to detect & quantitate ? 1) Precipitation: Ouchterlony 2) Solid-state binding, labelled antibody: RIA/ELISA 4) Consumption of complement: Complement Fixation (read-out is hemolysis) 3) Liquid-phase binding: Equilibrium Dialysis ...see simulator on web site. HTf HTf HSA BSA Ab complete identity non- identity partial identity X X Y HSA BSA Ouchterlony Analysis: 2-Dimensional Precipitation in Agarose Appendix 3 Ab = -whole human serum Ab HSA Ab HSA Ab HSA Ab HSA Ab HSA Ab HSA Ouchterlony: Double Diffusion in Agarose Ab = rabbit anti-HSA Reversible binding; lines may move by dissolving and re-precipitating Ouchterlony: Double Diffusion in Agarose HSA Ab BSA HSA Ab BSA Ab HSA BSA Ab HSA BSA Ab HSA BSA Ab HSA BSA X, Y Y X X Y HSA BSA Ab = rabbit anti-HSA...
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This note was uploaded on 01/16/2012 for the course BI 200 taught by Professor Potter during the Fall '11 term at Montgomery College.

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LEC04PH - 1 Immunology 2011 Lecture 4 Antibody Structure II...

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