通过实验设计,加速开发一种SEC-HPLC程序,用于单克隆抗体的纯度分析
Terezie Cernosek1, Nitin Jain2, Matthew Dalphin2
1Catalent Biologics, Madison, WI, USA; Keck Graduate Institute of Applied Life Sciences, Claremont, CA, USA.
概括
优化分析尺寸排除染色体 (SEC) 提高了单克隆抗体 (mAb) 尺寸变异的量化. 新的方法提高了低分子量 (LMW) 物种的分辨率,并增加了高分子量 (HMW) 物种的恢复.
科学领域:
- 生物制药学分析
- 分析化学 分析化学
- 蛋白质的表征 蛋白质的表征
背景情况:
- 由于其复杂的结构,生物制药产品需要严格的表征.
- 分析尺寸排除染色学 (SEC) 对于评估单克隆抗体 (mAbs),量化关键质量属性 (CQAs),如聚合物和碎片至关重要.
- 现有的SEC方法由于具有相似的分子质量而难以解决低分子量 (LMW) 物种,影响了准确的量化.
研究的目的:
- 开发一个强大的分析SEC方法,以更好地量化mAb大小变量.
- 优化SEC列类型和移动相组合,以更好地分辨LMW物种.
- 为了提高高分子量 (HMW) 物种的恢复.
主要方法:
- 使用实验设计 (DoE) 方法系统评估SEC参数.
- 用于测试方法的稳定性,使用了两种具有明显生理化学性质的mAb产品.
- 研究了各种SEC列类型和移动相组合,包括L-氨酸等添加剂.
主要成果:
- 使用WatersTM BioResolve列的优化SEC程序显示了增强的分辨率,特别是对于LMW物种.
- 优化的方法提高了准确量化mAb尺寸变异的能力.
- 添加L-氨酸作为移动相添加剂减少了二次相互作用,并增加了HMW物种的恢复.
结论:
- 开发的分析SEC方法提供了改进的分辨率和mAb大小变异的量化,解决LMW物种分析的局限性.
- 使用L-氨酸对减少二次相互作用和改善HMW恢复有好处.
- 这种优化的SEC方法有助于更好地表征和质量控制mAb疗法.
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