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通过构建轻链二级库的方法 ,对人源抗 TNF-α单抗 Fab进行轻链置换 ,并筛选出具有更高亲和力的人源抗 TNF- α单抗的 Fab.首先用 RT- PCR技术扩增正常人全套抗体轻链基因 ,并与已获得的人源抗 TNF- α单抗的重链基因配对 ,构建人源抗 TNF- α噬菌体抗体轻链二级库 ,然后筛选与 TNF- α具有更高亲和力的克隆 .经过三轮的生物淘筛 ( biopanning) ,获得了比原来的人源抗 TNF-α单抗 Fab具有更高亲和力的人源抗体 Fab.轻链二级库筛选法能够有效使抗体的亲和力提高 ,为解决噬菌体抗体库法制备的人源抗体的亲和力较低这一难题提供了一种有效的方法 .
By constructing a light chain secondary library, the human anti-TNF-α monoclonal antibody Fab was subjected to light chain replacement and the Fab with higher affinity of the human anti-TNF- α monoclonal antibody was screened.First, RT-PCR The full-length light chain gene of normal human was amplified and paired with the obtained heavy chain gene of human anti-TNF-α monoclonal antibody to construct the secondary anti-TNF-α phage antibody light chain library. After three rounds of biopanning, a human antibody Fab with a higher affinity than the original human anti-TNF- [alpha] monoclonal antibody Fab was obtained The light chain secondary library screening method was able to Effectively increasing the affinity of the antibody provides an effective method to solve the problem of the low affinity of the human antibody prepared by the phage antibody library method.