A new matrix for affinity chromatography and its application in the separation of a human monoclonal antibody

A method for the preparation of a p-aminobenzene sulphonyl ethyl containing crosslinked Sepharose 4B (ABSE-Sepharose 4B-CL) is described. trypsin, bovine serum albumin (BSA) and concanavalin A (Con A) were immobilized onto this matrix by diazotization. Conditions for the coupling reaction were investigated. The activity of immobilized trypsin reached 1.4 × 104U/g, and 25 mg BSA can be coupled onto 1 g ABSE-Sepharose 4B-CL under the optimal conditions. An affinity medium with immobilized Con A as ligand was prepared by this method, and was used in the separation of a human monoclonal antibody.