Antibody variable region interactions with Protein A: Implications for the development of generic purification processes
Antibody variable region interactions with Protein A: Implications for the development of generic purification processes
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DOI：
10.1002/bit.20729
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发表时间：
2005-12-20
影响因子：
3.8
通讯作者：
Cramer, SM
中科院分区：
文献类型：
--
作者：
Ghose, S;Allen, M;Cramer, SM
在本文中，来自各种亚类和亚家族的广泛的抗体被用来评估使用蛋白A层析的通用分离过程的创建。使用几种互补技术研究了蛋白A色谱过程中几种IgG1、IgG2、抗体片段和Fc融合蛋白之间洗脱pH值差异的原因。结果表明，可变区相互作用在使用传统的蛋白A色谱材料时确定V（H）3亚家族抗体的洗脱pH中起主要作用。另一方面，使用采用仅由蛋白A的B结构域组成的配体的树脂的实验表明可变区相互作用可以减轻，使得能够对一系列抗体使用单一洗脱pH。最后，与该工程化配体相关的洗脱条件的缓和显示出与低pH诱导的聚集相关的问题最小化。预计本文中报告的研究结果将促进这类重要的人类疗法的更快的工艺开发周期。(c)2005 Wiley Periodicals，Inc.
In this paper, a wide range of antibodies from various subclasses and subfamilies are employed to evaluate the creation of generic separation processes using Protein A chromatography. The reasons for elution pH differences amongst several IgG1s, IgG2s, antibody fragments, and Fc-fusion proteins during Protein A chromatography are investigated using several complimentary techniques. The results indicate that variable region interactions play a major role in determining elution pH for V(H)3 subfamily antibodies while using traditional protein A chromatographic materials. On the other hand, experiments with a resin which employs a ligand consisting solely of B domain of Protein A indicate that variable region interactions can be mitigated, enabling the use of a single elution pH for a range of antibodies. Finally, the moderation of elution conditions associated with this engineered ligand are shown to minimize problems associated with low pH induced aggregation. It is expected that the findings reported in this paper will facilitate faster process development cycle times for this important class of human therapeutics. (c) 2005 Wiley Periodicals, Inc.