Generation of Antibody Libraries for Phage Display: Preparation of Helper Phage.

Cold Spring Harbor protocols Pub Date : 2024-10-15 DOI:10.1101/pdb.prot108600

Haiyong Peng, Christoph Rader

引用次数: 0

Abstract

The generation and selection of antibody libraries by phagemid-based phage display requires three components; namely, phagemid library, host bacterial cells, and helper phage. The use of helper phage is necessary for the selection of phagemid libraries by phage display because it provides all genes needed for production of infectious phage particles. Here, we describe the generation of high-titer helper phage preparations suitable for phagemid-based phage display. The approach is based on helper phage VCSM13, which includes a gene for kanamycin resistance and a mutated packaging signal that, in the presence of a phagemid with an unmutated packaging signal, favors the production of infectious phage particles with phagemid phenotype and genotype.

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生成用于噬菌体展示的抗体库：制备辅助噬菌体

利用基于噬菌体的噬菌体展示技术生成和筛选抗体文库需要三个组成部分，即噬菌体文库、宿主细菌细胞和辅助噬菌体。使用辅助噬菌体是通过噬菌体展示筛选噬菌体文库的必要条件，因为辅助噬菌体提供了产生感染性噬菌体颗粒所需的所有基因。在此，我们介绍了适合噬菌体展示的高滴度辅助噬菌体制备方法。这种方法以辅助噬菌体 VCSM13 为基础，VCSM13 包括一个卡那霉素抗性基因和一个变异的包装信号，在带有未变异包装信号的噬菌体存在的情况下，有利于产生具有噬菌体表型和基因型的传染性噬菌体颗粒。

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.

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