The identification of antigens in antibody/antigen complexes using high performance liquid chromatography

Abstract

A method for the identification of antigens in a complex mixture of proteins is described. It exploits the effect which addition of a monoclonal antibody and resultant formation of antibody/antigen complexes has on the apparent retention time of the antigen on a size exclusion column. An antigen in the mixture can be identified as that component whose retention time is decreased by addition of its antibody. The principle of the method is demonstrated using an HPLC size exclusion column (DuPont GF450) and three MoAbs, PD7/26, DA6 147 and W6/32 which are directed against three antigens of the lymphocyte surface, the leucocyte-common antigen, class II MHC and class I MHC respectively.