不同的VH3结合蛋白A树脂表现出相似的VH3结合中介副产品分离能力,尽管它们对VH3 Fab具有不同的动态结合能力
Lixia Hu1, Rongrong Wang1, Qinxue Wu1
1Department of Downstream Process Development (DSPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai 200131, China.
Protein and peptide letters
|August 20, 2024
概括
不同的蛋白A树脂表现出基于VH3域结合的抗体副产品分离的类似能力,即使结合能力不同. 这为净化具有相同Fc区域但不同VH3域数量的抗体提供了可行的策略.
科学领域:
- 生物制药净化 生物制药净化
- 蛋白质染色学 蛋白质染色学
- 抗体工程 抗体工程
背景情况:
- 蛋白A树脂对于通过结合Fc区域来净化单克隆抗体 (mAbs),双特异抗体 (bsAbs) 和Fc融合蛋白至关重要.
- 一些蛋白A配体也与VH3域结合,使具有较少VH3域但相同Fc区域的副产品分离.
- 不同的VH3结合蛋白A树脂,来自不同的蛋白A域,可能提供不同的分离能力.
研究的目的:
- 研究各种VH3结合蛋白A树脂在分离抗体物种中的潜力.
- 为了比较不同树脂在区分抗体产品和截断的副产品的有效性,基于VH3域价值.
主要方法:
- 从mAbs中通过papain消化将VH3 Fab片段分离出来,并使用亲和色谱进行净化.
- 五种VH3结合蛋白A树脂的动态结合能力 (DBC) 用纯化的VH3 Fab.进行了评估.
- 使用人工混合产品 (一个VH3域) 和副产品 (零VH3域) 来评估分离能力,两者共享相同的Fc区域.
- 二甲基硫酸盐-聚烯胺凝电泳 (SDS-PAGE) 用于监测净化和分离.
主要成果:
- 个别的VH3结合蛋白A树脂在装载VH3 Fab时表现出不同的动态结合能力.
- 尽管存在DBC变异,但所有测试的蛋白质A树脂在分离具有相同Fc区域但不同VH3域号码的抗体物种方面表现出相似的有效性.
- 具体来说,将一个VH3域的产品从一个缺乏VH3域的副产品中分离出来,在树脂中以类似的方式实现.
结论:
- 虽然与VH3结合的蛋白A树脂在与VH3 Fab的结合能力上有所不同,但它们在根据VH3域含量分离抗体物种时的性能相似.
- 这表明VH3结合蛋白A染色学是一种强大的方法,用于净化具有相同Fc区域但具有不同的VH3域价值的抗体.
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