Rapid optimization of immunoadsorbent characteristics.
Immunoaffinity chromatography is employed in many research areas. We have developed an assay system 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 sulphate. Non-covalently bound proteins are dissociated and separated during the electrophoresis from IA beads. A wide variety of associative and dissociative conditions can be tested on small amounts of IA. Information about biospecifially 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.
D000936:Antigen-Antibody Complex; D002846:Chromatography, Affinity / Q000379:methods*; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007163:Immunosorbent Techniques; D007164:Immunosorbents; D009994:Osmolar Concentration
D000936:Antigen-Antibody Complex; D003902:Detergents; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007164:Immunosorbents