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使用丝状噬菌体展示进行亲和选择。

Affinity selection using filamentous phage display.

作者信息

Kulseth Mari Ann, Fagerlund Annette, Myrset Astrid Hilde

机构信息

Department of Medical Genetics, Rikshospitalet, Oslo University Hospital, Oslo, Norway.

出版信息

Methods Mol Biol. 2014;1088:67-80. doi: 10.1007/978-1-62703-673-3_5.

Abstract

Display of peptides on filamentous phage, phage display, is an in vitro selection technique well suited for identification of therapeutic peptide binders for a huge variety of protein targets. The peptides are identified in a process where phage libraries are subjected to affinity selection towards a particular protein target. A successful outcome of an affinity selection is dependent on proper surveillance of the phage life cycle, to make sure that the selection is based on affinity for the target, not on bias in phage propagation rate. In this chapter we present two approaches for protein target presentation and a protocol for phage rescue and propagation, which includes several controls to ensure that all phages initially eluted from the protein target are given equal conditions during the following amplification and selection steps.

摘要

在丝状噬菌体上展示肽,即噬菌体展示,是一种体外筛选技术,非常适合用于鉴定针对多种蛋白质靶标的治疗性肽结合物。在噬菌体文库针对特定蛋白质靶标进行亲和选择的过程中鉴定这些肽。亲和选择的成功结果取决于对噬菌体生命周期的适当监测,以确保选择基于对靶标的亲和力,而不是基于噬菌体繁殖速率的偏差。在本章中,我们介绍了两种蛋白质靶标呈递方法以及噬菌体拯救和繁殖方案,其中包括几个对照,以确保从蛋白质靶标最初洗脱的所有噬菌体在后续扩增和选择步骤中获得平等条件。

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