Rapid optimization of immunoadsorbent characteristics

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Abstract
Immunoaffinity chromatography is employed in many research areas. An assay system was developed that overcomes some of the tediousness and uncertainty in dealing with immunoadsorbents (IA). The preparation of IA and the effect of various procedures on the dissociation of antigen-antibody complex and the regeneration of IA are rapidly screened by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulfate. Non-covalently bound proteins are dissociated and separated from IA beads during the electrophoresis. A wide variety of associative and dissociative conditions can be tested on small amounts of IA. Information about biospecifically and non-biospecifically adsorbed proteins can also be obtained. By using relatively small volumes of media containing either an antigen or another biospecific molecule, the optimal parameters for affinity chromatography (specificity, binding capacity, efficiency of solvents in dissociation of the complex and their effect on the adsorbent), or even for ion-exchange chromatography, can be determined without first performing several time- and material-consuming chromatographic experiments.