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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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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 x 10~5 independent clones). We first constructed the Ve cDNA library (4.6 x 10~5 independent clones) and V kappa cDNA library (3.0 x 10~5 independent clones). Then, the V epsilon and V kappa gene 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 V epsilon 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,构建了鼠类IgE噬菌体表面展示文库(3.0 x 10〜5个独立的克隆)。我们首先构建了Ve cDNA文库(4.6 x 10〜5个独立克隆)和V kappa cDNA文库(3.0 x 10〜5个独立克隆)。然后,通过PCR分别从两个文库中扩增出V epsilon和V kappa基因片段,并通过SOE PCR组装成Fab片段。由此构建了包含Fab的噬菌体文库。分析并证明了该库中V epsilon的多样性。已筛选出对TCS具有高度特异性的Fab克隆。

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