天然鼠源噬菌体抗体库的构建及抗羊抑制素基因工程单抗的筛选

Construction of Naive Mouse Phage Antibody Library and Screening of Monoclonal Antibodies Specific for Sheep Inhibin

【目的】构建单链抗体(ScFv)噬菌体表面展示文库,从中筛选抗羊抑制素单抗并进行表达,建立制备羊抑制素单抗的新方法。【方法】用未经免疫的多种小鼠脾细胞作为基因来源,采用噬菌体抗体库技术,构建天然噬菌体表面抗体文库。用羊抑制素对其进行3轮吸附-洗脱-富集,筛选抗羊抑制素单抗,用ELISA检测其抗原结合活性。【结果】构建了天然鼠源噬菌体抗体库,为筛选和制备各种单链抗体提供了一个平台。用羊抑制素对其进行筛选,制备抗羊抑制素单抗,经ELISA检测,55/96克隆具有与羊抑制素结合活性,阳性率为57%。【结论】制备的羊抑制素可溶性ScFv及其表面展示噬菌体抗体有很好的抗原结合活性,为羊抑制素单抗的制备提供了一种新的方法。

[Objective] The purpose of this study is to construct the naive murine phage antibody library and acquire monoclonal antibodies against sheep inhibin by screening. [Method] The naive murine phage antibody library was constructed using phage display technology. The library was selected by using sheep inhibin as panning antigen by 3 rounds of binding-elution-enrichment procedure. The binding ability of expressed ScFv was detected by ELISA. [ Result] The naive murine phage antibody library was established. Monoclonal antibodies against sheep inhibin were acquired by screening. Fifty-five out of 96 randomized clones were demonstrated to have binding capacities to sheep inhibin, with the positive ratio as much as 57%. [Conclusion] The expressed ScFv had the ability to bind sheep inhibin. The production of genetic engineering antibody against sheep inhibin has provided a basis for further study. This strategy may provide an alternative newer approach to antibodies.