【摘要】 【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。更多还原

【Abstract】 【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.更多还原