治疗性单克隆抗体─抗药抗体 亲和常数 确定 使用毛细管电泳-双重质谱 质谱学
Tessa Reinert1,2, Pascal Houzé2,3, Nathalie Mignet2
1Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), Université de Strasbourg, CNRS UMR7140, CMC, Strasbourg 67081, France.
Analytical chemistry
|November 27, 2024
概括
我们开发了一种新的毛细电泳质谱法来测量抗药抗体亲和力. 该技术准确量化了单克隆抗体 (mAb) 和患者血清中的抗药抗体相互作用的解离常数 (Kd).
科学领域:
- 生物制药学分析分析
- 免疫学 免疫学 免疫学
- 分析化学是一种分析化学.
背景情况:
- 单克隆抗体 (mAb) 疗法可以引起免疫反应,导致抗药抗体 (ADA).
- ADAs可以中和mAbs,降低治疗效果并加速清除.
- 了解mAb-ADA相互作用对于预测生物治疗结果至关重要.
研究的目的:
- 引入一种新的分析方法来确定mAbs和ADAs之间的离散常数 (Kd).
- 用患者血清在生理条件下测量mAb-ADA亲和力.
- 为了验证该方法的准确性和稳定性,用于高亲和度相互作用.
主要方法:
- 毛细管电泳与质谱学 (CE-MS/MS) 结合的开发和应用.
- 使用竞争性试验量化残留活性mAbs.
- 采用质量摄影法 (MP) 进行蛋白质-蛋白质相互作用的初始表征和固体测量.
主要成果:
- 成功测量Kd的infliximab和一个中和抗infliximab抗体为14.4±2.9nM.
- 证明了在没有蛋白质固定化的情况下测量纳米分子范围内的高 afinity 相互作用的能力.
- 验证了CE-MS/MS方法的灵敏度和特异性,用于在血清中稳定地确定Kd.
结论:
- 新的CE-MS/MS方法为在原生血清条件下测量mAb-ADA亲和力提供了一个可靠的平台.
- 这种方法有助于更深入地了解生物治疗行为和潜在的免疫性.
- 该技术适用于特征强蛋白-蛋白相互作用相关的治疗抗体监测.
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