【摘要】 从Tomlinson(I+J)噬菌体抗体库中筛选获得了特异性抗GST蛋白的单链抗体(scFv),并在E.coliHB2151进行表达,经SDS-PAGE和Western blot分析表明,scFv可在细菌周质腔中进行可溶性表达,其分子量约为30 kD,与理论预期值相符;ELISA鉴定结果显示,周质腔中的scFv具有抗原结合功能,而且这种结合可以被GST纯蛋白所抑制,其IC50值约为3.07 ng/ml,证明该scFv具有特异性结合GST蛋白的活性。本研究表明,利用噬菌体抗体库技术可以不经过免疫就能获得高特异性的人源化抗GST蛋白的scFv。更多还原

【Abstract】 A novel human anti-GST single-chain antibodies(scFv) was cloned from Tomlinson(I+J) phage antibody library,then expressed in the E.coli BL21(DE3).Results obtained from SDS-PAGE and Western blot assay demonstrated that scFv with molecular weight of 30 kD was expressed in the bacterial periplasm,and the analysis with ELISA showed that the titer of scFv in the inperiplasmic extracts was 1:16 and the scFv in bacterial periplasm could competitively combine with GST,where IC50 was 3.03 ng/ml.These results indicate that highly specific humanized anti-GST single antibody can be obtained without immunization by using a phage display antibody library technique.更多还原