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

Antigen-Antibody Interactions

Dr. Conrad

Sept. 10,2008


Antigenicity

Antigenicity

  • Requirements to be an immunogen

  • Low mw antigens (haptens)

  • Adjuvants

  • Primary vs secondary responses


Antigen antibody interactions

Concept of a Hapten


Antigen antibody interactions

r/c = Kn-Kr

c = concentration of free antigen (hapten)

K = Ka = equilibrium constant

r = hapten bound per unit Ab

n = Valence of Ab


Antigen antibody interactions

Polyclonal Ab


Antigen antibody interactions

1/c = K(2) – K(1) 1/c = K0


Factors which influence ab affinity

Factors which influence Ab affinity

  • Temp, pH, ionic strength

  • Polyclonal vs monoclonal

  • Ab class

  • Type of antigen


Antibody combining site

Antibody Combining Site

Location


Antigen antibody interactions

All interactions are non-covalent


Evidence that hv interacts with ag

Evidence that Hv interacts with Ag

3 lines of evidence


Antigen antibody interactions

Affinity Labeling


Antigen antibody interactions

Comparison of Hv sequences of monoclonals binding the

Same vs binding a different antigen epitope


Antigen antibody interactions

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.


Antigen antibody interactions

IgM is the most avid of Ab classes


Antigen antibody interactions

Composition of immune ppts at different Ab/Ag ratios


Antigen antibody interactions

Gel Diffusion Analysis to determine relatedness

of antigens


Antigen antibody interactions

Example of a determinant that depends on secondary structure


Antigen antibody interactions

ELISA

RIA


Antigen antibody interactions

Affinity Chromatography


Antigen antibody interactions

Agglutination


Antigen antibody interactions

Immunoprecipitation


Antigen antibody interactions

Western Blotting


Antigen antibody interactions

Immunofluorescence


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