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 x 10(4) U/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.
Guo
W
W
Dalian Institute of Chemical Physics, Academia Sinica, China.
Shang
Z
Z
Yu
Y
Y
Zhou
L
L
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D003208:Concanavalin A; D004800:Enzymes, Immobilized; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D012685:Sepharose; D012710:Serum Albumin, Bovine; D013424:Sulfanilamides; D014357:Trypsin
D000911:Antibodies, Monoclonal; D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D013424:Sulfanilamides; D003208:Concanavalin A; C089193:4-((2-sulfatoethyl)sulfonyl)aniline; D012710:Serum Albumin, Bovine; D012685:Sepharose; D014357:Trypsin