Arora Sushrut, Saxena Vikas, Ayyar B Vijayalakshmi
Department of Pediatrics, Baylor College of Medicine, Houston, TX 77030, USA.
Center for Vascular and Inflammatory Diseases, School of Medicine, University of Maryland, Baltimore, MD 21201, USA.
Methods. 2017 Mar 1;116:84-94. doi: 10.1016/j.ymeth.2016.12.010. Epub 2016 Dec 22.
Antibodies continue to be extremely utilized entities in myriad applications including basic research, imaging, targeted delivery, chromatography, diagnostics, and therapeutics. At production stage, antibodies are generally present in complex matrices and most of their intended applications necessitate purification. Antibody purification has always been a major bottleneck in downstream processing of antibodies, due to the need of high quality products and associated high costs. Over the years, extensive research has focused on finding better purification methodologies to overcome this holdup. Among a plethora of different techniques, affinity chromatography is one of the most selective, rapid and easy method for antibody purification. This review aims to provide a detailed overview on affinity chromatography and the components involved in purification. An array of support matrices along with various classes of affinity ligands detailing their underlying working principles, together with the advantages and limitations of each system in purifying different types of antibodies, accompanying recent developments and important practical methodological considerations to optimize purification procedure are discussed.
抗体在众多应用中仍然是极为常用的物质，这些应用包括基础研究、成像、靶向递送、色谱分析、诊断和治疗。在生产阶段，抗体通常存在于复杂的基质中，并且它们的大多数预期应用都需要进行纯化。由于对高质量产品的需求以及相关的高成本，抗体纯化一直是抗体下游加工中的主要瓶颈。多年来，广泛的研究集中在寻找更好的纯化方法以克服这一阻碍。在众多不同的技术中，亲和色谱是抗体纯化最具选择性、快速且简便的方法之一。本综述旨在详细概述亲和色谱以及纯化过程中涉及的组件。讨论了一系列支持基质以及各类亲和配体，详述了它们的基本工作原理，以及每个系统在纯化不同类型抗体时的优缺点，同时还介绍了近期的进展以及优化纯化程序的重要实际方法学考量。
