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混合模式树脂在抗体纯化中的应用。

Application of mixed mode resins for the purification of antibodies.

机构信息

Institute for Chemical and Bioengineering, Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland.

出版信息

J Chromatogr A. 2010 Sep 10;1217(37):5753-60. doi: 10.1016/j.chroma.2010.06.047. Epub 2010 Jun 25.

Abstract

The downstream processing of monoclonal antibodies from cell culture supernatant is usually done by a number of chromatographic and non-chromatographic steps. Efforts are taken to reduce the costs associated to those steps, while maintaining a high product purity. A possibility to reach this goal is the reduction of the number of chromatographic steps using mixed mode resins that offer more than one functionality in one chromatographic step. In this work, a commercially available mixed mode resin was evaluated systematically with respect to the adsorption of proteins. The Henry coefficient, which quantifies the adsorption strength, was measured for the full working range of the stationary phase as a function of the salt concentration and the pH. The results were compared to a conventional anion exchange and a hydrophobic interaction resin. Furthermore, the resin was applied for the polishing step of an antibody from an industrial clarified cell culture supernatant.

摘要

单克隆抗体的细胞培养上清液下游处理通常需要经过多步色谱和非色谱步骤。人们努力降低这些步骤的相关成本,同时保持高的产品纯度。实现这一目标的一种可能性是使用混合模式树脂来减少色谱步骤的数量,因为这种树脂在一个色谱步骤中提供了多种功能。在这项工作中,对一种商业上可用的混合模式树脂进行了系统评估,以研究其对蛋白质的吸附能力。亨利系数定量了吸附强度,它被测量为固定相的整个工作范围的函数,与盐浓度和 pH 值有关。结果与传统的阴离子交换树脂和疏水相互作用树脂进行了比较。此外,该树脂还应用于从工业澄清细胞培养上清液中进行抗体的抛光步骤。

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