摘要
以Sepharose CL-4B-Pro A吸附胃癌单克隆抗体(McAb)PD4,继之以交联剂二甲基庚二亚胺二盐酸盐(dimethyl pimelimidate dihydrochloride)处理,形成亲和介质(柱Ⅰ)。该亲和介质用于纯化抗PD4独特型抗体(aIdAb)具有明显的优点。与Sepharose CL-4B-PD4(柱Ⅱ)相比,前者的结合容量为后者的1.78～1.94倍。采用柱Ⅰ纯化的aIdAb,当其与McAb PD4的分子比分别为1:1及4:1时可50％或100％地抑制McAb PD4与靶细胞的结合,但采用柱Ⅱ获得的aIdAb,只有当分子比达到2:1及8:1时,才能达到同等的抑制效应。这可能是由于Pro A与McAb PD4的Fc片断结合,使后者的Fab端得以充分暴露,因而有更多的机会与aIdAb结合。
Monoclonal antibody (McAb) PD4 against gastric cancer was absorbed on Sepharose CL-4B-Pro A and then treated with crosslinking agent dimethyl pimelimi date dihydrochloride to form affinity medieum Sepharose CL-4B-Pro A-PD4 (columnⅠ). Column I had obvious advantages in Purifing anti-PD4-idiotypic antibody (αIdAb) . over the Sepharoae CL-4B-PD4 column (column Ⅱ).The binding capacity of columnⅠ with αIdAb was 1.78-1.94 times of that of column n .And the αIdAb purified with column Ⅰ had higher ability to inhibit in combination of PD4 with target cells than that of column Ⅱ . When the ratio of aldAb from column I to McAb PD4 was 1:1 and 4:1, the inhibition could reach 50% and 100% respectively, whereas for aldAb purified with column Ⅱ, the ratio of 2:1 and 8:1 was needed to exhibit same inhibition effects. Because of the protein A may combine with the Fc fragments of McAb PD4 to expose the PD4 Fab fragmets, therefore it is possible to provide PD4 with more opportunity for combining with aldAb.
基金
国家自然科学基金
关键词
抗独特型抗体
亲和层析
Anti-idiotypic antibody, Affinity chromatography
