Generation of a Phage Display Chicken Single-Chain Variable Fragment Library.

Cold Spring Harbor protocols Pub Date : 2024-08-16 DOI:10.1101/pdb.prot108213

Hyunji Yang, Jisu Chae, Hyori Kim, Jinsung Noh, Junho Chung

引用次数: 0

Abstract

Phage-displayed antibody fragment libraries can be constructed using essentially any species that is easily immunized, as long as the immunoglobulin variable region gene sequences are known. This protocol describes the procedures for the generation of a phage-displayed chicken single-chain variable fragment (scFv) library after immunization with a target antigen. Briefly, the rearranged heavy chain variable region (V _H ) genes and the λ light chain variable region (V _λ ) genes are amplified separately and are linked through two separate PCR steps to give the final scFv genes. The genes are then cloned into pComb3XSS to generate the phage display chicken scFv library, which can then be used for test and final library ligations.

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噬菌体展示鸡单链可变片段库的生成。

只要知道免疫球蛋白可变区基因序列，噬菌体显示抗体片段文库基本上可以用任何容易免疫的物种来构建。本方案介绍了用目标抗原免疫后生成噬菌体显示的鸡单链可变片段（scFv）文库的程序。简言之，分别扩增重链可变区（V H）基因和 λ 轻链可变区（V λ）基因，并通过两个独立的 PCR 步骤连接，得到最终的 scFv 基因。然后将这些基因克隆到 pComb3XSS 中，生成噬菌体展示鸡 scFv 文库，该文库可用于测试和最终文库连接。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cold Spring Harbor protocols Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)

CiteScore

3.00

自引率

0.00%

发文量

163

期刊介绍： Cold Spring Harbor Laboratory is renowned for its teaching of biomedical research techniques. For decades, participants in its celebrated, hands-on courses and users of its laboratory manuals have gained access to the most authoritative and reliable methods in molecular and cellular biology. Now that access has moved online. Cold Spring Harbor Protocols is an interdisciplinary journal providing a definitive source of research methods in cell, developmental and molecular biology, genetics, bioinformatics, protein science, computational biology, immunology, neuroscience and imaging. Each monthly issue details multiple essential methods—a mix of cutting-edge and well-established techniques.