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链霉亲和素包被固相萃取(SPE) tips 用于抗体噬菌体展示生物淘选。

Streptavidin-Coated Solid-Phase Extraction (SPE) Tips for Antibody Phage Display Biopanning.

机构信息

Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Penang, Malaysia.

出版信息

Methods Mol Biol. 2023;2702:275-290. doi: 10.1007/978-1-0716-3381-6_14.

Abstract

Phage display is a technique that allows the presentation of unique proteins on the surface of bacteriophages. The phage particles are usually screened via repetitive rounds of antigen-guided selection and phage amplification. The main advantage of this approach lies in the physical linkage between phenotype and genotype. This feature allows the isolation of single unique clones from a panning campaign consisting of a highly diverse population of clones. Due to the high-throughput nature of this technique, different approaches have been developed to assist phage display selections. One of which involves utilizing a streptavidin-coated solid-phase extraction (SPE) tip that is mounted to an electronically controlled motorized multichannel pipette. In this chapter, we will entail the procedures involved in the adaptation of a commercial SPE tip (MSIA™ streptavidin D.A.R.T's) as the solid phase. This protocol is an updated version of a previous protocol with some minor refinements.

摘要

噬菌体展示是一种将独特蛋白质展示在噬菌体表面的技术。噬菌体颗粒通常通过抗原导向的选择和噬菌体扩增的重复轮次进行筛选。这种方法的主要优点在于表型和基因型之间的物理联系。该特征允许从由高度多样化的克隆群体组成的淘选活动中分离单个独特的克隆。由于该技术具有高通量的特点,因此已经开发出不同的方法来辅助噬菌体展示选择。其中一种方法涉及使用涂有链霉亲和素的固相萃取 (SPE) 吸头,该吸头安装在电子控制的多通道移液器上。在本章中,我们将详述将商用 SPE 吸头(MSIA™链霉亲和素 D.A.R.T's)适作为固相的过程。该方案是对之前方案的更新版本,有一些小的改进。

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