通过CE-SDS监测低分子量蛋白质聚合,作为SE-HPLC的补充方法
Si-Tao Wang1, Min-Fei Sun2, Han Gao2
1Institute of Drug Metabolism and Pharmaceutical Analysis, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
Journal of pharmaceutical and biomedical analysis
|June 16, 2023
概括
用二硫酸盐 (CE-SDS) 进行毛细电泳,可以有效地表征低分子量蛋白质和,甚至可以识别特定的胰岛素格拉聚合物,而这种聚合物被大小排除高性能液体染色学 (SE-HPLC) 遗漏. 这扩大了生物制药分析中的CE-SDS应用.
科学领域:
- 生物制药学分析
- 蛋白质的表征 蛋白质的表征
- 分析化学 分析化学
背景情况:
- 用二硫酸盐 (CE-SDS) 进行毛细电泳,用于治疗性蛋白质分析.
- 它对低分子量蛋白 (<10kDa) 和的应用较少被探索.
- 精确的蛋白质聚合物的表征对于药物的安全性和有效性至关重要.
研究的目的:
- 证明CE-SDS对于低分子量蛋白和多的纯度评估的实用性.
- 评估CE-SDS在压力条件下 (热量,光线) 鉴定胰岛素格拉金聚合物的特征.
- 为了将CE-SDS与大小排除高性能液体染色学 (SE-HPLC) 进行比较,进行聚合分析.
主要方法:
- 使用二硫酸盐 (CE-SDS) 的毛细管电泳被使用.
- 胰岛素glargine样本受到热和光诱导的降解.
- 质谱法用于总体确认.
- 使用大小排除高性能液体染色学 (SE-HPLC) 进行比较分析.
主要成果:
- CE-SDS成功地分离了胰岛素格拉基因单体,二元体和三元体.
- 质谱学证实了两种类型的胰岛素聚合物,而SE-HPLC显示只有一种.
- 变化条件通过CE-SDS.显示只有共价聚合物.
- 与SE-HPLC相比,CE-SDS提供了更详细的汇总信息.
结论:
- CE-SDS 是一种强大的工具,用于描述低分子量蛋白和多的纯度.
- 与SE-HPLC相比,CE-SDS在检测和区分蛋白质聚合物方面提供了更高的分辨率.
- 在生物制药分析中,CE-SDS是SE-HPLC的宝贵补充技术.
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