Purification of Human Monoclonal Antibodies and Their Fragments.
Purification of Human Monoclonal Antibodies and Their Fragments.
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人单克隆抗体及其片段的纯化。
DOI：
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发表时间：
2018
影响因子：
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通讯作者：
M. Morbidelli
中科院分区：
文献类型：
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作者：
Nicole Ulmer;Sebastian Vogg;T. Müller;M. Morbidelli
本章总结了最常见的色谱mAb和mAb片段纯化方法，首先阐明化合物的相关性质，并介绍了可用于此应用的各种色谱模式。重点放在捕获步骤亲和和离子交换色谱。可扩展性方面在判断方法的适用性方面起着重要作用。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。在mAb的情况下，对于大多数目的，使用亲和捕获步骤获得的纯度是足够的。如果需要产生特别高纯度的材料，则需要抛光步骤。对于mAb片段，亲和色谱尚未完全建立，捕获步骤可能无法提供高纯度的材料。因此，对现有的抛光技术作了简要的介绍。就mAb亚型和双特异性抗体纯化而言，逆流色谱技术已被证明非常有用，本章的一部分专门讨论了它们，向研究和工业中对这些抗体形式日益增长的兴趣致敬。
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 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.