用单克隆抗体亲和层析法纯化重组人肿瘤坏死因子α  

Purification of Recombinant Human Tumor Necrosis Factor Alpha by Affinity Chromatography with Monoclonal Antibodies

作　　者：许辉[1] 安献禄[2] 刘雪松[1] 金伯泉[1] 王俊龙[2] 智刚[2] 黄传书[1] 

机构地区：[1]第四军医大学免疫学教研室,西安710032 [2]第四军医大学药物研究所

出　　处：《免疫学杂志》1992年第2期87-89,共3页Immunological Journal

摘　　要：用抗rHuTNF-α单抗(G_(11)、D_(12))与溴化氰活化的Sepharose4B偶联制成亲和层析柱,纯化了rHuTNF-α。结果表明,经G_(11)-Sepharose4B柱纯化的TNFα比活性比纯化前提高2.71～4.51倍;经D_(12)-Sepharose4B柱纯化的TNF-α比活性提高2.3～3.4倍。G_(11)-Sepha-rose4B柱的蛋白回收率高于D_(12)-Sepharose4B柱。纯化产物经SDS-PAGE确定达电泳纯,其分子量约为17000道尔顿。The recombinant human tomur necrosis factor Alpha (rHuTNF-α) was purified with affinity chromatography colunms which were prepared by binding monoclonal antibodies (G_(11), D_(12)) against rHuTNF-α to CNBr-activated Sepharose 4B. It was showed that the biological activitiesof G_(11)-Sepharose 4B-purified rHuTNF-α and D_(12)-Se pharose 4B-purified that increased 2.71~4.51 and 2.3~3.4 fold respectively in comparison with pre-affinity chromatography crude lysates. The rHuTNF-α recovery rate of Gu-Sepharose 4B was higher than that of D_(12)-Sepharose 4B and the purified rHuTNF-α was showed as a single band with a 17KD molecular weight on SDS-PAGE.

关 键 词：肿瘤坏死因子-A 亲和层析 单克隆抗体 

分 类 号：R730.3[医药卫生—肿瘤]