抗体的上下和中下质谱分析
Nina A Khristenko1, Konstantin O Nagornov2, Camille Garcia3
1Spectrotech, 69006 Lyon, France.
Molecular & cellular proteomics : MCP
|May 14, 2025
概括
本综述强调了30年来对分析治疗抗体的上下和中下质谱学的进展. 这些质谱法 (MS) 技术确保为抗体表征保留了关键质量属性.
科学领域:
- 生物化学 生化学
- 分析化学 分析化学
- 免疫学 免疫学 免疫学
背景情况:
- 治疗性抗体,主要是单克隆抗体,对于治疗癌症和自身免疫性疾病等疾病至关重要.
- 它们的复杂性质带来了重大的分析挑战,需要先进的表征方法.
- 顶向下 (TD) 和中向下 (MD) 质谱 (MS) 已经成为抗体分析的关键技术.
研究的目的:
- 审查TD和MDMS的30年演变,以进行治疗性抗体分析.
- 突出关键的技术进步和该领域的持续挑战.
- 讨论这些MS技术对新型抗体模式的应用.
主要方法:
- 顶向下 (TD) 质谱仪用于完整的抗体分析.
- 中向下 (MD) 质谱法用于抗体子单元分析.
- 使用先进的离子激活方法 (例如,ETD,ECD,UVPD,MALDI-ISD) 和高分辨率质谱仪.
主要成果:
- 通过使用激活离子ETD,TD MS可以实现高达60%的完整抗体序列覆盖率.
- 通过组合激活和解离技术,MDMS可以达到抗体子单元的多达95%的序列覆盖率.
- 无论是TD还是MDMS都保持了关键的质量属性,包括序列完整性,PTM,二硫化键和糖化.
结论:
- 在过去的三十年中,TD和MD MS在抗体表征方面取得了重大进展.
- 这些技术对于分析复杂的抗体结构和新型模式,如抗体-药物联合体和双特异性抗体至关重要.
- 持续开发MS方法对于全面的抗体分析和质量控制至关重要.
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