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Updated: Jan 24, 2026

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使用镜像蛋白酶素和Tryp-NN的单克隆抗体的LC-MS基映射

Domantas Sargautis¹, Bernd Thiede¹

¹Department of Biosciences, University of Oslo, Oslo, Norway.

Journal of pharmaceutical and biomedical analysis

|January 22, 2026

概括

在单克隆抗体 (mAb) 分析中,Tryp-N提供了与素相似的序列覆盖. 两种酶的结合略有改善了覆盖范围,并有助于精确的翻译后修饰 (PTM) 定位.

科学领域:

生物化学生物化学
分析化学分析化学
生物制药产品生物制药产品

背景情况:

对单克隆抗体 (mAbs) 的准确表征对于产品质量评估至关重要.
通过自下而上的液体染色体质谱法 (LC/MS) 绘制片映射是一种标准技术.
素消化为mAbs.提供高但不完整的序列覆盖.

研究的目的:

评估Tryp-N作为mAb基映射的替代性内蛋白酶.
为了比较Tryp-N与素的性能.
评估补充序列在PTM局部化中的有用性.

主要方法:

四个mAbs的自下而上的LC/MS分析 (贝瓦齐祖马布, cetuximab,NISTmAb, trastuzumab).
使用素,Tryp-N和两者的组合进行酶性消化.
对序列和翻译后修改 (PTM) 的分析.

主要成果:

在所有测试的mAbs.中,Tryp-N实现了与素相当的序列覆盖率.
结合素和Tryp-N略有提高了整体序列覆盖范围.
两种酶都产生了相同的,除了末端,从而实现了互补的覆盖.
来自相同的序列的补充离子序列促进了精确的PTM定位 (除化,氧化,SAVs).

结论:

在mAb序列的特征化中,Tryp-N是一种可行的替代品.
酶组合可以在片映射中提高序列覆盖率.
补充分析可以提高PTM和单氨基酸变体的局部精度.

关键词:

镜子蛋白质酶是一种镜子蛋白质酶.单克隆抗体是一种单克隆抗体.没有一个NISTmAbAb.类映射映射类映射后翻译修改后的修改.单氨基酸变体的单氨基酸变体这是Tryp-NN.

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