宿主细胞蛋白质丰富的聚合物在抗体捕获和抛光染色学中的行为
Chase E Herman1, Lie Min1, Leila H Choe1
1Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE 19716, USA.
Journal of chromatography. A
|May 27, 2023
概括
单克隆抗体 (mAb) 聚合物,通常含有宿主细胞蛋白 (HCP),经过净化而持续存在. 总质量分数与HCP水平相关,这表明它可以作为过程开发中的HCP清除策略的替代品.
科学领域:
- 生物制药制造业 生物制药制造业
- 蛋白质聚合 蛋白质聚合
- 主体细胞蛋白 (HCP) 清除 清除
背景情况:
- 单克隆抗体 (mAb) 聚合物可以含有宿主细胞蛋白 (HCP).
- 聚合物通过净化持续存在可能会影响HCP移除效率.
研究的目的:
- 在关键宿主细胞蛋白减少步骤中调查聚合行为.
- 为了确定总质量分数和净化过程中的HCP水平之间的关系.
主要方法:
- 通过深度过,蛋白A染色学和离子交换 (AEX) 染色学分析聚合物持久性.
- 同焦激光扫描显微镜观察聚合物-mAb相互作用.
- 与酶相关的免疫吸收试验 (ELISA) 和蛋白质组分析用于HPC量化.
主要成果:
- 聚合物与mAbs在蛋白质A色谱中的结合点上竞争,影响洗效率.
- 蛋白A化物和AEX流通分数显示出显著的总质量分数 (分别为2.4-3.6%和1.5-3.2%).
- 总体质量分数与测量的HCP度和识别的HCP正相关.
结论:
- 聚合物持久性在多个净化步骤中是相关的,包括深度过,蛋白A和AEX染色学.
- 含有HCP的聚合物可以通过AEX染色学保留,这取决于树脂化学.
- 量化总质量分数可以作为一个有用的,虽然不完美的,替代品来评估HCP清除在早期的过程开发.
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