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电荷检测质谱揭示了巨型克隆抗体聚合物的结构异质性

Jacob S Jordan¹, Conner C Harper¹, Fan Zhang²

¹Department of Chemistry, University of California, Berkeley, California 94720-1460, United States.

Journal of the American Chemical Society

|August 7, 2024

概括

电荷检测质谱 (CDMS) 有效地表征大抗体聚合物及其构造,克服了蛋白质聚合研究的传统方法的局限性.

科学领域:

生物化学
分析化学
生物技术

背景情况:

蛋白质聚合是治疗开发的关键问题,影响疗效和安全性.
通过标准分析技术,对大型抗体聚合物及其构成进行鉴定是具有挑战性的.

研究的目的:

评估用于分析大型单克隆抗体 (mAb) 聚合物的电荷检测质谱法 (CDMS).
将CDMS与本地MS,SEC和CE-SDS等常规方法进行比较,以进行总体表征.

主要方法:

使用原生质谱法 (MS) 分析pentameric mAb复合物.
使用电荷检测质谱法 (CDMS) 来检测和描述单个寡合物和更高阶聚合物.
使用尺寸排除色谱 (SEC) 和毛细电泳SDS (CE-SDS) 进行比较分析.

主要成果:

CDMS 分解了个别的mAb寡合体,其质量达到了约2.4 MDa (五合体mAb三合体).
传统方法无法检测或解决中间聚合物和更高阶物种.
由于充电范围的差异,CDMS发现了mAb寡合体的独特构造,这些构造无法通过其他技术辨认.

结论:

对于大抗体聚合物的精确质量确定和构造分析,CDMS具有显著的优势.
CDMS可以研究早期蛋白质聚合物的结构变化,这对于理解聚合途径至关重要.

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