[小鼠可变免疫球蛋白基因免疫组合cDNA文库的构建与鉴定]

[Construction and characterization of immune combinatorial cDNA library of mouse variable immunoglobuline genes].

作者信息

Pavlova M V, Hil'chuk P V, Pokholenko Ia O, Nikolaiev Iu S, Kordium V A

出版信息

Tsitol Genet. 2008 Mar-Apr;42(2):10-5.

Abstract

A cDNA combinatorial antibody library of mouse variable immunoglobulin fragments has been constructed from mice immunized with rhIFN-beta1b. For this purpose, cDNAS of immunoglobulin variable heavy (V(H)) and variable light (V(L)) chains genes amplified from splenocytes were joined with linker DNA to form ScFv's (single-chain Fv-antibodies). The obtained ScFv-DNA pool was cloned into a phagemid vector and used for Esherichia coli transformation. Using the phage display technique, bacterial clones producing single-chain antibodies specific to rhIFN-beta1b were selected. The following characteristics of the combinatorial library were determined in this work: abundance, functional size, and the initial ScFv-DNA diversity in the library constructed. High specificity of interaction between phage displayed ScFv's and rhIFN-beta1b has been demonstrated.

摘要

用重组人干扰素β1b免疫小鼠后构建了小鼠可变免疫球蛋白片段的cDNA组合抗体文库。为此，从脾细胞中扩增的免疫球蛋白重链可变区（V(H)）和轻链可变区（V(L)）基因的cDNA与接头DNA连接形成单链抗体（ScFv）。将获得的ScFv-DNA文库克隆到噬菌粒载体中并用于大肠杆菌转化。利用噬菌体展示技术，筛选出产生对重组人干扰素β1b具有特异性的单链抗体的细菌克隆。在本研究中确定了该组合文库的以下特性：丰度、功能大小以及所构建文库中初始ScFv-DNA的多样性。已证明噬菌体展示的ScFv与重组人干扰素β1b之间具有高度特异性的相互作用。