通过增加交叉流梯度来增加场流分化的分辨率
Nicholas R Larson1, George M Bou-Assaf1
1Pharmaceutical Operations & Technology, 225 Binney Street, Cambridge, Massachusetts 02142, United States.
Analytical chemistry
|October 24, 2023
概括
这项研究引入了一种新型的梯度方法,用于在分离过程中增加交叉流速的流域-流量分成 (流量FFF). 这种技术可以提高复杂样品的峰值分辨率,例如单克隆抗体和病毒.
科学领域:
- 分析化学 分析化学
- 分离科学 分离科学
背景情况:
- 在流域-流量分化 (流FFF) 中的分辨率主要由交叉流速控制.
- 传统方法在分离过程中保持恒定或下降的交叉流速.
研究的目的:
- 开发和验证一种新的流量FFF梯度方法,该方法调节交叉流速以提高分离分辨率.
- 为了证明这种方法在不同的FFF技术和样本类型中是否适用.
主要方法:
- 实施了梯度方法,在分离过程中短暂增加交叉流速.
- 在单克隆抗体单体和聚合物之间的量化分辨率改进.
- 将该方法应用于非对称流动FFF (AF4) 和用单克隆抗体和腺相关病毒进行水力动力色谱 (HF5).
主要成果:
- 与传统方法相比,新型梯度方法显著提高了峰值至峰值分辨率.
- 已在AF4和HF5中成功应用,改善了复杂的生物样本的分离.
- 验证了该方法在分离单克隆抗体变异和病毒载体方面的有效性.
结论:
- 通过在分离过程中增加交叉流速来调节交叉流速,这是一个广泛适用的策略,以提高所有FFF方法的解决方案.
- 这种方法为基于FFF的挑战性样本分离提供了更好的质量.
- 梯度方法为后期排泄的峰值提供了更强的保留,从而提高了整体分离效率.
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