Haba S, Nisonoff A
Department of Biology, Brandeis University, Waltham, MA 02254.
J Immunol Methods. 1991 Apr 8;138(1):15-23. doi: 10.1016/0022-1759(91)90059-o.
Conditions are described for the preparation of F(ab')2 and Fab fragments of mouse IgE. Papain, pepsin or trypsin each produced F(ab')2 fragments with Mr approximately equal to 130,000 which yielded Fab fragments on further digestion. The release of Fab fragments from F(ab')2 resulted from further cleavage of the H chain. Pepsin, and especially trypsin appear more suitable for the preparation of F(ab')2 because of the difficulty of separating a 93 kDa by-product from the F(ab')2 produced by papain. The best yields of purified Fab were obtained with papain. Rates of digestion were in the order, pepsin approximately equal to trypsin much greater than papain.
描述了制备小鼠IgE的F(ab')2和Fab片段的条件。木瓜蛋白酶、胃蛋白酶或胰蛋白酶均可产生Mr约为130,000的F(ab')2片段,进一步消化后可产生Fab片段。F(ab')2中Fab片段的释放是由于重链的进一步裂解。由于难以从木瓜蛋白酶产生的F(ab')2中分离出93 kDa的副产物,胃蛋白酶,尤其是胰蛋白酶似乎更适合制备F(ab')2。用木瓜蛋白酶可获得最高产量的纯化Fab。消化速率依次为:胃蛋白酶≈胰蛋白酶>>木瓜蛋白酶。