Arora Sushrut, Ayyar B Vijayalakshmi, O'Kennedy Richard
National Centre for Sensor Research, Biomedical Diagnostics Institute, Dublin City University, Dublin, Ireland.
Methods Mol Biol. 2014;1129:497-516. doi: 10.1007/978-1-62703-977-2_35.
The availability of purified antibodies is prerequisite for many applications and the appropriate choice(s) of antibody-purification steps is crucial. Numerous methods have been developed for the purification of antibodies; however, affinity chromatography-based methods are the most extensively utilized. These methods are based on highly specific and reversible biological interactions between two molecules (e.g., between receptor and ligand or antibody and antigen). Affinity chromatography offers very high selectivity, involving minimal steps, providing simplicity of approach and rapidity. Implementing an effective protocol often requires meticulous planning and testing in order to achieve high purity and yields of desired antibody types/subtypes. This chapter describes the basic techniques for purification of monoclonal, polyclonal, and recombinant antibodies employing affinity chromatography.
获得纯化抗体是许多应用的前提条件，而正确选择抗体纯化步骤至关重要。已经开发出许多用于纯化抗体的方法；然而，基于亲和色谱的方法应用最为广泛。这些方法基于两个分子之间高度特异性和可逆的生物相互作用（例如，受体与配体之间或抗体与抗原之间）。亲和色谱具有非常高的选择性，步骤最少，方法简便且快速。要实施有效的方案，通常需要精心规划和测试，以实现所需抗体类型/亚型的高纯度和高产量。本章介绍了使用亲和色谱纯化单克隆抗体、多克隆抗体和重组抗体的基本技术。
