Chap.6–Antigen-Antibodyinteractions.pptVIP

Chap. 6 – Antigen-Antibody interactions Characterized as: Non-covalent interaction (similar to “lock and key” fit of enzyme-substrate) Does not lead to irreversible alteration of Ag or Ab This exact and specific interaction has led to many immunological assays used to: detect Ag or Ab diagnose disease measure magnitude of humoral IR identify molecules of bio and med interest Ag-Ab interactions Bonds: Hydrogen Ionic Hydrophobic interactions Van der Waals forces Each bond is weak; many are strong To “hold” they must be close ? requiring high amts of complementarity! Measuring affinity of Ab to Ag Ka determined by equilibrium dialysis Cross-reactivity Immunologic tests: Immunologic tests: Immunologic tests: 2. Immunoelectrophoresis: Incorp electrophoresis w/ double diffusion An Ag mixture is 1st separated by charge Then, “troughs” are cut ∥to direction of elec field and antisera is added to trough Ag’s and Ab’s diffuse towards each other to produce precipitin bands Used to detect: a)presence/absence of specific proteins or Ig classes b) immunodeficiency or immunoproliferative disorder Immunologic tests: Immunologic tests: 3) Agglutination reactions – simple, inexpensive, but sensitive! Several types exist: a) Hemagglutination of RBC’s b) Bacterial Agglutination c) Passive Agglutination d) Agglutination Inhibition Immunologic tests: 4) Radioimmunoassay (RIA)– very sensitive test; used for measuring hormones, serum proteins, drugs, etc. at low [C]’s (≤ 0.001ug/ml) measures “competitive binding” of radiolabelled Ag + unlabelled (test) Ag to high affinity Ab Immunologic tests: Immunologic tests: Types of ELISA’s… Immunologic tests:  6) Western Blot Used to id specific proteins in mixtures Proteins are separated on SDSProteins then transferred to membrane Membrane flooded w/ radio-labelled or enz-linked poly/monoclonal Ab’s specific for protein Immunologic tests: 7) Immunoprecipitation Provides a