Müller-Späth Thomas, Morbidelli Massimo
ETH Zurich, Institute for Chemical and Bioengineering, Zurich, Switzerland.
Methods Mol Biol. 2014;1060:331-51. doi: 10.1007/978-1-62703-586-6_17.
This chapter summarizes the most common chromatographic mAb and mAb fragment purification methods, starting by elucidating the relevant properties of the compounds and introducing the various chromatography modes that are available and useful for this application. A focus is put on the capture step affinity and ion exchange chromatography. Aspects of scalability play an important role in judging the suitability of the methods. The chapter introduces also analytical chromatographic methods that can be utilized for quantification and purity control of the product. In the case of mAbs, for most purposes the purity obtained using an affinity capture step is sufficient. Polishing steps are required if material of particularly high purity needs to be generated. For mAb fragments, affinity chromatography is not yet fully established, and the capture step potentially may not provide material of high purity. Therefore, the available polishing techniques are touched upon briefly. In the case of mAb isoform and bispecific antibody purification, countercurrent chromatography techniques have been proven to be very useful and a part of this chapter has been dedicated to them, paying tribute to the rising interest in these antibody formats in research and industry.
本章总结了最常见的单克隆抗体（mAb）和单克隆抗体片段纯化方法，首先阐述了这些化合物的相关特性，并介绍了适用于此应用的各种色谱模式。重点放在捕获步骤的亲和色谱和离子交换色谱上。可扩展性方面在判断这些方法的适用性中起着重要作用。本章还介绍了可用于产品定量和纯度控制的分析色谱方法。对于单克隆抗体，在大多数情况下，使用亲和捕获步骤获得的纯度就足够了。如果需要生成特别高纯度的材料，则需要进行精制步骤。对于单克隆抗体片段，亲和色谱尚未完全确立，捕获步骤可能无法提供高纯度的材料。因此，简要介绍了可用的精制技术。在单克隆抗体异构体和双特异性抗体纯化的情况下，逆流色谱技术已被证明非常有用，本章的一部分专门介绍了这些技术，以回应研究和工业界对这些抗体形式日益增长的兴趣。
