Question

can immunoelectrophoresis provide an indication of the relative concentration of antigen-specific antibodies? why and why not ?

Answer 1

Immunoelectrophoresis is a powerful qualitative technique for the characterization of any antibody. In this method one antigen mixture is electrophoresed in an agarose gel that allows the separation of its different components based on their charge along the gel side, followed by the lateral diffusion of the serum or monoclonal antibody within the gel. Antibodies specific to the antigens form white precipitation arcs which can be seen against a dark background. This technique is useful in determining the blood levels of immunoglobins.

The antigen molecules migrate at different rates through the electric current because of differing electric charges at a specific pH value. The most common procedures in food analysis rely upon the migration of antigenic proteins through or into an antibody- containing gel. Immunoelectrophoresis provide an indication by RID assay is similar to the Ouchteroomy assay but is used to precisely quantify antigen concentration rather than to compare different antigens. The square of the diameter of the zone of precipitation made by the antigen antibody interact is directly proportional to the concentration of antigen. By measuring the zone of precipitation produced by samples of known concentration , we can prepare a standard curve for determining the concentration of specific antigen-antibody solution.

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