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亲和标签在蛋白质纯化和肽富集中的应用:概述。

Affinity Tags in Protein Purification and Peptide Enrichment: An Overview.

机构信息

UCIBIO, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Caparica, Portugal.

Nanoscience Centre, Department of Engineering, University of Cambridge, Cambridge, UK.

出版信息

Methods Mol Biol. 2021;2178:107-132. doi: 10.1007/978-1-0716-0775-6_10.

Abstract

The reversible interaction between an affinity ligand and a complementary receptor has been widely explored in purification systems for several biomolecules. The development of tailored affinity ligands highly specific toward particular target biomolecules is one of the options in affinity purification systems. However, both genetic and chemical modifications in proteins and peptides widen the application of affinity ligand-tag receptors pairs toward universal capture and purification strategies. In particular, this chapter will focus on two case studies highly relevant for biotechnology and biomedical areas, namely the affinity tags and receptors employed on the production of recombinant fusion proteins, and the chemical modification of phosphate groups on proteins and peptides and the subsequent specific capture and enrichment, a mandatory step before further proteomic analysis.

摘要

亲和配体与互补受体之间的可逆相互作用已在多种生物分子的纯化系统中得到广泛探索。针对特定靶生物分子的定制亲和配体的开发是亲和纯化系统的选择之一。然而,蛋白质和肽的遗传和化学修饰拓宽了亲和配体-标签受体对的应用范围,使其成为通用捕获和纯化策略的选择。特别是,本章将重点介绍两个在生物技术和生物医学领域非常相关的案例研究,即用于重组融合蛋白生产的亲和标签和受体,以及蛋白质和肽上磷酸基团的化学修饰及其随后的特异性捕获和富集,这是进一步进行蛋白质组学分析之前的必需步骤。

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