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碰撞诱导的展开允许在复杂的生物矩阵中对同质生物疗法进行量化

Brock R Juliano¹, Brandon T Ruotolo¹

¹Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States.

Journal of the American Society for Mass Spectrometry

|August 16, 2023

概括

量化碰撞诱导展开 (CIU) 质谱法使同质蛋白质的无标签量化成为可能. 这种方法成功地在复杂的生物样本中区分和量化蛋白质异型,为生物治疗分析提供了一个新的工具.

科学领域:

分析化学分析化学
生物化学生物化学
质谱测量质量谱测量

背景情况:

定量质谱 (MS) 通常依赖于质量解析的离子,这对量化异构或形状上不同的分析物提出了挑战.
准确量化同质量蛋白质,共享相同的质量电荷比,对于生物治疗分析和药物开发至关重要.

研究的目的:

开发和验证量化碰撞诱导展开 (CIU) 质谱法,用于对同质蛋白质的无标签量化.
证明CIU-MS在各种矩阵中区分和量化结构相似的蛋白质异型的能力.

主要方法:

使用原生离子移动性质谱 (nIM-MS) 与碰撞诱导展开 (CIU) 相结合.
CIU利用碰撞激活来诱导展开,产生不同的气相结构来分离蛋白质异型.
应用了定量CIU方法来测量同质生物治疗对和序列同类的度.

主要成果:

该CIU-MS方法成功地解决和量化了标准和生物矩阵中的同质抗体对和序列同类.
校准曲线显示出高线性,相关系数在0.92到0.99.9之间.
该方法实现了敏感检测,检测极限从300到5000nm,灵敏度从8.7×10−5nM−1到6×10−3μM−1不等.

结论:

定量CIU-MS是一种强大的技术,用于无标签量化异质蛋白质,包括生物疗法.
该方法在复杂混合物中区分和量化蛋白质异型的能力对制药分析和个性化医学具有重大前景.
预计CIU启用定量MS的未来应用将用于先进的生物制药特征和质量控制.

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