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分离蛋白质度和聚合物含量使用扩散和水NMR

Mark I Grimes¹, Matthew Cheeks², Jennifer Smith²

¹Department of Chemical Engineering and Biotechnology, University of Cambridge, Philippa Fawcett Drive, Cambridge CB3 0AS, U.K.

Analytical chemistry

|June 29, 2024

概括

水的NMR (wNMR) 现在可以分离蛋白质度和聚合水平. 通过测量水扩散 (D) 与放松率 (R2) 一起,该方法为生物过程监测提供了一个新的工具.

科学领域:

生物制药学分析分析
过程分析技术 (PAT) 是一种分析技术.
核磁共振 (NMR) 光谱学核磁共振 (NMR) 光谱学

背景情况:

基于蛋白质的药物,如单克隆抗体,是全球领先的制药品.
监测蛋白质度和聚合物水平对于生物过程控制至关重要.
使用横向放松率 (R2) 的水NMR (wNMR) 可以测量蛋白质度和聚合物,但无法区分它们.

研究的目的:

开发一种方法来"解"并同时确定生物制药中的蛋白质度和总量水平.
克服现有的wNMR技术的局限性,即无法区分这两个关键参数.

主要方法:

使用的水扩散系数 (D ((H2O)) 测量与水横向放松率 (R2 ((H2O)) 来自wNMR.
在三个不同的蛋白质系统上演示了该方法.
涉及到特定于感兴趣的蛋白质的D (H2O) 和R (H2O) 的校准数据采集.

主要成果:

成功解并同时确定蛋白质度和聚合物水平.
结合D ((H2O) 和R2 ((H2O) 的方法即使在单个参数重叠的情况下也有效.
在各种蛋白质系统中验证了该方法.

结论:

组合的wNMR方法 (D ((H2O) + R2 ((H2O)) 有效地分离了蛋白质度和总量水平.
这种技术显示了作为在线过程分析技术的实施前景.
能够更精确地监测和控制基于蛋白质的治疗方法的生物过程.

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