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在已构建的抗结肠癌相关抗原单链抗体基因的基础上,为方便其活性测定,设计了含有E tag的引物进行PCR扩增,以scFv-E tag融合基因的形式构建在质粒pET-22b(+)中,并在大肠杆菌中进行了表达。对表达产物的主要成分包涵体进行了变性、复性,ELISA及免疫组化SABC法对该单链抗体进行活性测定,结果表明它具有与原代单克隆抗体相似的抗原结合活性及组织特异性。
Based on the established single-chain antibody against colon cancer antigen gene, an E-tagged primer was designed for PCR amplification and the scFv-E tag fusion gene was constructed for expression in the plasmid pET-22b (+) And expressed in E. coli. The inclusion bodies of the major components of the expressed product were denatured and renatured. The activity of the single chain antibody was determined by ELISA and immunohistochemical SABC. The results showed that the inclusion body had similar antigen-binding activity and tissue specificity .