含有E tag的抗结肠癌相关抗原单链抗体的研制

Construction and Activity Analysis of Anticolonic Cancer scFv Fragment Containing E tag

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。

In order to detect the activity of anticolonic cancer scFv fragment, a primer containing E tag was designed for PCR amplification. The fusion gene scFv-E tag was cloned to plasmid pET-22b ( + )and was expressed in E. coli and the inclusion body of the main expression products was denatured and renatured. Activity analysis showed that the scFv fragment had the similar antigen affinity and specificity to the parent monoclonal antibody by using ELISA and immunochemi-cal methods.