Muronets V I, Cherednikova T V, Haggadova N K
Biokhimiia. 1976 Nov;41(11):2081-5.
A method of Fab fragments preparation by enzymatic splitting of antibodies bound to specific antigen immobilized on an insoluble support is described. The complex of rat muscle glyceraldehyde-3-phosphate dehydrogenase (GAPD), immobilized on Sepharose 4B, with anti-rat GAPD rabbit antibodies was digested with papain. The antigen was inaccessible to proteolysis under conditions employed. After 4 hrs of incubation with papain the antibody was completely split into non-precipitating fragments. The products of proteolysis not bound to Sepharose, were eluted with 0.1 M givcine buffer pH 2.5, and shown to correspond to Fab fragments.
描述了一种通过酶解固定在不溶性支持物上的与特定抗原结合的抗体来制备Fab片段的方法。固定在琼脂糖4B上的大鼠肌肉甘油醛-3-磷酸脱氢酶(GAPD)与抗大鼠GAPD兔抗体的复合物用木瓜蛋白酶消化。在所采用的条件下,抗原不能被蛋白酶解。用木瓜蛋白酶孵育4小时后,抗体完全裂解为非沉淀性片段。未与琼脂糖结合的蛋白水解产物用pH 2.5的0.1M甘氨酸缓冲液洗脱,结果表明其对应于Fab片段。