增强的蛋白质沉与氨使得快速,通用提取的生物分析的寡核酸
Afrand Kamali1, Juili D Shelke1, Ethan J Sanford1
1Novo Nordisk Research & Development US Inc., 65 Hayden Ave, Lexington, Massachusetts 02421, United States.
ACS omega
|October 20, 2025
概括
这项研究引入了使用有机溶剂中的小氨基的增强蛋白沉 (EPP). EPP显著改善了生物样本中的寡核酸 (ONT) 恢复,提供了一种低成本,高效的生物分析方法.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 药理学 药理学是指药理学的学科.
背景情况:
- 氧核酸 (ONT) 的发现和开发需要有效的,通用的生物分析方法.
- 传统的蛋白质沉对于ONT由于恢复不良而失败.
- 现有的ONT提取方法往往是昂贵和复杂的.
研究的目的:
- 开发一种具有成本效益和效率的通用方法,从生物矩阵中提取寡核酸.
- 在生物分析过程中改善ONT的恢复.
- 建立适用于不同类型的寡核酸的多功能方法.
主要方法:
- 在有机溶剂中使用小氨基的增强蛋白沉 (EPP) 的开发.
- 对于反意义寡核酸 (ASOs) 和小干扰RNA (siRNAs) 的EPP的分析合格.
- 使用离子配对逆相 (IPRP) 液态染色学,并与双重质谱 (MS/MS) 和高分辨率MS (HRMS) 相结合.
主要成果:
- 从血和组织中,EPP实现了ASO和siRNA的80%以上的恢复.
- 对于各种ONT,可达到的定量化下限 (LLOQ) 为1-5 ng/mL.
- 已证明适用于多个ONT类,无需样本消化或专用板.
结论:
- EPP是一种多功能和可持续的寡核酸生物分析方法.
- 该方法消除了复杂的样本准备步骤的需要.
- 与传统的寡核酸提取技术相比,EPP提供了显著的进步.
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