SDS-PAGE after micro-affinity adsorption for analysis of heterogeneous antigen polypeptides in individual sera

A method was developed for the analysis of heterogeneity in antigen polypeptides in individual sera. Polypeptides in sera were adsorbed by polystyrene beads coated with antibody in wells of a microplate. They were dissociated with a small volume of elutant, and transferred to slots on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Polypeptides separated on gel were then immunoblotted with antibodies labeled with horseradish peroxidase. The method was applied to analyze different populations of hepatitis B surface and e antigen polypeptides in sera from carriers of hepatitis B virus. Applicability to mass-scale and high sensitivity of the method would allow surveys of heterogeneous antigen polypeptides in serum for any biological significance.