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目的制备抗重组GST的单克隆抗体(mAb), 并用来纯化重组GST融合蛋白。方法用含重组GST融合蛋白基因的pGEX4T -1质粒转化E.coli BL21，IPTG诱导GST融合蛋白表达，亲和层析和凝胶过滤法分离表达的重组GST融合蛋白。以此蛋白作为抗原，免疫Balb/c小鼠，按传统的杂交瘤技术制备mAb。将抗GST mAb经Protein A纯化后，与Sepharose 4B偶联。结果经3次亚克隆后，获得两株分泌抗GST载体特异性mAb的杂交瘤。采用该mAb对两种不同的GST融合蛋白进行亲和层析纯化后，经SDS-PAGE鉴定达到了商品化Glutathione-Resin的亲和层析纯化效果。结论用抗GST蛋白特异性mAb亲和层析纯化融合蛋白是一种经济、实用的方法，且可用于Glutathione-Resin亲和层析纯化后的二次纯化。
Objective To prepare monoclonal antibodies against recombinant GST (mAbs) and to purify recombinant GST fusion proteins. Methods E. coli BL21 was transformed with pGEX4T -1 plasmid containing recombinant GST fusion protein. The expression of GST fusion protein was induced by IPTG. The recombinant GST fusion protein was isolated by affinity chromatography and gel filtration. Using this protein as an antigen, Balb / c mice were immunized and mAb was prepared according to a conventional hybridoma technique. Anti-GST mAbs were purified by Protein A and coupled to Sepharose 4B. Results After three subclones, two hybridomas secreting anti-GST vector-specific mAbs were obtained. After purified by affinity chromatography of two different GST fusion proteins by this mAb, the affinity purification purification of commercial Glutathione-Resin was achieved by SDS-PAGE. Conclusion The purification of the fusion protein by affinity chromatography with anti-GST protein-specific mAb is an economical and practical method and can be used for the second purification after purified by Glutathione-Resin affinity chromatography.