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Antigen-Antibody Interactions

Antigen-Antibody Interactions. Dr. Conrad Sept. 10,2008. Antigenicity. Requirements to be an immunogen Low mw antigens (haptens) Adjuvants Primary vs secondary responses. Concept of a Hapten. r/c = Kn-Kr. c = concentration of free antigen (hapten) K = K a = equilibrium constant

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Antigen-Antibody Interactions

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  1. Antigen-Antibody Interactions Dr. Conrad Sept. 10,2008

  2. Antigenicity • Requirements to be an immunogen • Low mw antigens (haptens) • Adjuvants • Primary vs secondary responses

  3. Concept of a Hapten

  4. r/c = Kn-Kr c = concentration of free antigen (hapten) K = Ka = equilibrium constant r = hapten bound per unit Ab n = Valence of Ab

  5. Polyclonal Ab

  6. 1/c = K(2) – K(1) 1/c = K0

  7. Factors which influence Ab affinity • Temp, pH, ionic strength • Polyclonal vs monoclonal • Ab class • Type of antigen

  8. Antibody Combining Site Location

  9. All interactions are non-covalent

  10. Evidence that Hv interacts with Ag 3 lines of evidence

  11. Affinity Labeling

  12. Comparison of Hv sequences of monoclonals binding the Same vs binding a different antigen epitope

  13. Affinity vs avidity – depends on valence Avidity – sum total or strength of binding of two molecules (or cells) to one another at multiple sites. Distinct from affinity which is the strength of binding of one site on a molecule to its ligand.

  14. IgM is the most avid of Ab classes

  15. Composition of immune ppts at different Ab/Ag ratios

  16. Gel Diffusion Analysis to determine relatedness of antigens

  17. Example of a determinant that depends on secondary structure

  18. ELISA RIA

  19. Affinity Chromatography

  20. Agglutination

  21. Immunoprecipitation

  22. Western Blotting

  23. Immunofluorescence

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