Electroelution of antigens immobilized on antibody-linked affinity matrices.
An electrophoretic elution procedure for desorption of antigens from antibody-linked gel matrices is described which is performed in a commercially available elution device. The technique presented does not involve denaturing conditions such as chaotropic reagents or low and high pH elution buffers. Comparison of electroelution and conventional elution with acidic buffer reveals that the techniques is especially suitable for monoclonal antibodies with high affinity for their ligands. Recovery of antigens after electroelution is significantly higher than by desorption with glycine-HCl, pH 2.5. There is no loss of antigens during post-elution procedures. Proteins are obtained in small elution volumes and can be desalted without sample transfer.
Schulze-Osthoff
K
K
Institute of Experimental Dermatology, University of Münster, West Germany.
Michels
E
E
Overwien
B
B
Sorg
C
C
D043925:Angiogenesis Inducing Agents / Q000276:immunology / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000941:Antigens / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006133:Growth Substances / Q000302:isolation & purification*; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D013997:Time Factors
D043925:Angiogenesis Inducing Agents; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000941:Antigens; D006133:Growth Substances