Antigen antibody interactions
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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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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 = Ka = equilibrium constant

r = hapten bound per unit Ab

n = Valence of Ab


Polyclonal Ab


1/c = K(2) – K(1) 1/c = K0


Factors which influence Ab affinity

  • Temp, pH, ionic strength

  • Polyclonal vs monoclonal

  • Ab class

  • Type of antigen


Antibody Combining Site

Location


All interactions are non-covalent


Evidence that Hv interacts with Ag

3 lines of evidence


Affinity Labeling


Comparison of Hv sequences of monoclonals binding the

Same vs binding a different antigen epitope


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.


IgM is the most avid of Ab classes


Composition of immune ppts at different Ab/Ag ratios


Gel Diffusion Analysis to determine relatedness

of antigens


Example of a determinant that depends on secondary structure


ELISA

RIA


Affinity Chromatography


Agglutination


Immunoprecipitation


Western Blotting


Immunofluorescence


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