快速抗体结构特征和量化通过微滴滴素消化
Mengyuan Xiao1, Yongqing Yang1, Andrew Schladebeck1
1Department of Chemistry & Environmental Science, New Jersey Institute of Technology, Newark, New Jersey 07102, United States.
Journal of the American Society for Mass Spectrometry
|February 15, 2026
概括
我们开发了一种快速的方法来对抗体进行表征和量化,使用微滴素消化. 这种技术能够快速,准确地分析蛋白质修饰和单克隆抗体 (mAbs) 的绝对量化.
科学领域:
- 生物化学和蛋白质组学
- 分析化学 分析化学
- 生物制药学分析
背景情况:
- 蛋白质结构特征对于生物制药至关重要.
- 单克隆抗体 (mAbs) 需要快速准确的分析方法.
- 现有的蛋白质消化和量化方法可能耗时.
研究的目的:
- 开发一种快速的抗体表征和定量方法.
- 为了利用在线微滴素消化用于超快速蛋白质分析.
- 为了实现精确的片映射,修改定位和mAbs的绝对量化.
主要方法:
- 在线微滴素消化用于超快速的蛋白质消化 (<1 ms,>90%的效率).
- 质谱 (MS) 和MS/MS用于图和修改分析.
- 用标准和重同位素标记抗体进行绝对量化.
主要成果:
- 在NIST mAb中使用MS/MS.在N387确定了主要的脱化.
- 成功量化了使用绝对量化的氨酸氧化,并提高了准确度.
- 获得了敏感和准确的绝对抗体量化 (R2=0.99,LOD=1.2 ng,0.6-10%的误差).
结论:
- 微滴消化为抗体表征提供了一种快速有效的方法.
- 开发的方法提供了准确的翻译后修改和抗体水平的绝对量化.
- 这种技术对于生物制药分析非常有价值,它提供了速度,灵敏度和准确性.
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