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Construction and diversity analysis of a murine IgE phage surface display library

机译:鼠IgE噬菌体表面展示文库的构建和多样性分析

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摘要

To make further investigation of the IgE antibody repertoire in Trichosanthin (TCS) allergic responses,a murine IgE phage surface display library was constructed (3.0×105 independent clones).We first constructed the Vε cDNA library (4.6×105 independent clones) and Vκ cDNA library (3.0×105 independent clones).Then,the Vε and Vκgene segments were amplified from both libraries by PCR respectively,and assembled into Fab fragment by SOE PCR.The phage library containing Fabs was thus constructed.The diversity of Vε from this library was analyzed and proved.Fab clones with high specificity to TCS have been screened out.
机译:为了进一步研究天花粉蛋白(TCS)过敏反应中的IgE抗体库,构建了一个小鼠IgE噬菌体表面展示文库(3.0×105个独立克隆)。我们首先构建了VεcDNA文库(4.6×105个独立克隆)和Vκ cDNA文库(3.0×105个独立克隆),然后分别通过PCR从两个文库中扩增出Vε和Vκ基因片段,并通过SOE PCR将其组装成Fab片段,从而构建了包含Fab的噬菌体文库。分析并证明了文库。筛选出对TCS具有高度特异性的Fab克隆。

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