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通过毛细管区电泳量测量原生和强制降解的原蛋白:方法开发和验证

Mais Olabi¹, Hermann Wätzig¹

¹Institute of Medicinal and Pharmaceutical Chemistry, Technical University of Braunschweig, Braunschweig, Germany.

Journal of separation science

|October 26, 2023

概括

一种新的毛细管区电泳方法提供了一种快速和自动化的方法来量化原蛋白. 虽然它比Sircol试验的敏感度低,但它提供了更广泛的线性和对原蛋白完整性的洞察力.

科学领域:

分析化学分析化学
生物化学生化学
材料科学材料科学材料科学

背景情况:

在各种科学领域,原蛋白的量化是至关重要的.
像Sircol原蛋白试验这样的现有方法存在局限性.
需要改进,自动化原蛋白分析技术.

研究的目的:

开发和验证一种新的毛细管区电泳 (CZE) 方法来定量原蛋白.
将CZE的性能与已知方法 (如Sircol测定和紫外线光谱) 进行比较.
评估CZE对于分析原蛋白产品和降解样本的适用性.

主要方法:

开发和验证毛细管区电泳方法.
遵守国际协调理事会Q2 (R1) 准则进行验证.
使用Sircol原蛋白测定和紫外线光谱作为参考方法.

主要成果:

CZE提供了特定,简单和快速的原蛋白确定 (在9分钟内).
与Sircol测定相比,CZE对用户的依赖性较小,并且比Sircol测定更为自动化.
与Sircol (5-80μg/mL) 相比,CZE具有更广泛的线性范围 (50-400μg/mL).
CZE的检测极限比Sircol (6.5μg/mL) 高 (18.0μg/mL),但精度相似.
CZE提供了对原蛋白完整性和原生和变质原蛋白的同时确定的见解.

结论:

毛细管区电泳是一种现代和有效的替代品,用于原量化.
开发的CZE方法适用于研究原产品和强制降解样本.
与传统方法相比,CZE在速度,自动化和分析信息方面提供了显著的优势.

关键词:

锡尔科尔原蛋白测定试验毛细管区电泳是毛细管区的电泳.原蛋白的量化定量强制降解降解的强制性降解氧的确定方法

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