[Three-step purification of preparative-scale antiCD20 (Fab')2]

To establish a three-step purification method of preparative-scale antiCD20 (Fab')2 using AKTA prime.AntiCD20 (Fab')2 was extracted by hyperosmotic solution and then purified by CM sepharose FF, phenyl sepharose FF, and protein G sepharose FF.Around 8 mg anti-CD20 (Fab')2, whose purification was 96.678%, was purified. The antigen-binding activity of antiCD20 (Fab')2 was similar to that of antiCD20 (Fab')2 purified by protein G sepharose FF and S-100.The three-step purification method can obtain high-purity preparative-scale antiCD20 (Fab')2 in a simple way.