一个单克隆抗体的特征通过原生和变性自上而下的质谱学
Ryan N Oates1, Linda B Lieu1, Kristina Srzentić2
1Department of Chemistry and Biochemistry, University of Oklahoma, Norman, Oklahoma 73019, United States.
Journal of the American Society for Mass Spectrometry
|August 6, 2024
概括
原生质谱法可以对完整的单克隆抗体进行气相测序,比大多数碎片化技术的变质化方法提供更高的序列覆盖率. 电子转移解离对于特征抗体区域特别有效.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 质谱测量质量谱测量
背景情况:
- 原生质谱正在出现,用于分析单克隆抗体 (mAb) 异质性.
- 顶向下质谱 (MS/MS) 用于 mAb 测序通常使用变质条件.
- 有限的研究已经探索了原生MS的完整的mAb测序.
研究的目的:
- 为了对四种离子解离方法进行基准测试,以表征特斯图祖马布,一种治疗 mAb.
- 为了比较mAbs.的自上向下MS/MS测序的原生类和变性条件.
- 评估原生质谱法对于完整的mAb测序的实用性.
主要方法:
- 对高能碰撞解离 (HCD),电子转移解离 (ETD),ETD/HCD和UV光解离 (UVPD) 的系统基准测试.
- 对治疗性单克隆抗体trastuzumab的分析.
- 从变质和原生类前体离子的碎片化模式的比较.
主要成果:
- 原生自上而下的MS/MS实现了比变质条件更高的序列覆盖率,除了UVPD.
- 电子转移解离 (ETD),随后是基于碰撞的激活,在被二硫化物保护的区域产生了最多的骨干裂变,包括互补性确定区域 (CDR).
- 变质条件提供了一些在本地条件下没有观察到的脊柱裂纹.
结论:
- 原生质谱是一种可行的方法,用于完整的单克隆抗体的气相测序.
- 二硫化物键被认为是影响原生条件和变性条件之间重叠的解离模式的关键因素.
- 原生MS/MS为全面的抗体表征提供了强大的替代方案.
关键词:
EThcD 在线观看轨道上的陷电子转移解离,ETD,电子转移解离.高能碰撞解离,HCD,高能碰撞解离.一个单克隆抗体,mAb原生质谱法本土质谱法.从上到下,从上到下.紫外线光解离,UVPD,紫外线光解离.更多相关视频
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