多维LC-MS具有1D多方法选项和平行中向上和下向上MS获取,用于对抗体的深入表征
Liesa Verscheure1, Shauni Detremmerie2, Hilde Stals3
1RIC group, President Kennedypark 6, 8500 Kortrijk, Belgium; Separation Science Group, Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281-S4, 9000 Ghent, Belgium.
一种新型的自动化蛋白质分析仪使单克隆抗体变体的同时进行中向上和下向上液体染色体质谱 (LC-MS) 分析成为可能. 这个平台有效地描述电荷,大小和疏水性属性,用于全面的生物制药分析.
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:
- 单克隆抗体 (mAbs) 是复杂而异质的,需要多种分析方法进行表征.
- 生物制药行业正在向集成分析平台迈进,用于同时评估结构属性.
研究的目的:
- 描述一个完全自动化的蛋白质分析仪,用于中向上和下向上LC-MS分析.
- 为了能够同时评估mAbs.的电荷,大小和疏水变体.
主要方法:
- 一个多维LC-MS设置,将1D染色学 (SEC,CEX或HIC) 与2D列式RP-LC相结合,用于淡化,变质和减少.
- 3D柱上素消化,然后进行4DLC-MS进行自下而上的分析.
- 补充-交换 (HDX) MS和FcRn亲和力染色学-本土MS.
主要成果:
- 描述的蛋白质分析仪自动化了中上和自下而上的LC-MS工作流.
- 该系统成功地在一个工程Fc片段中表征了电荷,疏水和尺寸异质性.
- 综合分析方法为观察到的变异提供了结构和功能背景.
结论:
- 开发的多维LC-MS平台为mAb表征提供了多功能和全面的工作流.
- 该自动化系统解决了对生物制药多种结构属性的同时评估的需求.
- 该方法有助于理解mAb异质性的结构和功能影响.
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