Immunoaffinity chromatography

Abstract The principle of immunoaffinity or immunoadsorption chromatography is based on the highly specific interaction of an antigen with its antibody (1). Immunoaffinity chromatography is a specialized form of affinity chromatography and as such utilizes a ligand (either antigen or antibody) immobilized onto a solid support matrix in a manner which retains its binding capacity. Although the technique includes the separation of antibodies using immobilized antigens (2-4), it is more commonly performed for the identification, quantification, or purification of antigens (Figure 1). The crude extract is pumped through the column and the unbound material washed clear prior to elution of the retained antigen by alterations to the mobile phase conditions which weaken the antibody-antigen interaction.