改善酸净化中的分析和可转移性:从HPLC到FPLC,然后再回到HPLC
Alessandro Streuli1, Vanessa Erckes1, Brunello Nardone2
1Institute of Pharmaceutical Sciences, Laboratory of Pharmaceutical Analytics, ETH Zurich, Zurich, Switzerland.
概括
通过优化净化方法,实现高纯度的合成. 这项研究增强了分析高性能液体染色学 (HPLC) 转移到准备性快速蛋白质液体染色学 (FPLC) 进行可靠的净化.
科学领域:
- 生物化学 生化学
- 分析化学 分析化学
- 化学工程是化学工程的重要组成部分.
背景情况:
- 合成的高纯度对于生物活动和下游应用至关重要.
- 由于系统的变化,将分析性高性能液体染色学 (HPLC) 方法转移到准备性快速蛋白质液体染色学 (FPLC) 具有挑战性.
- 现有的方法缺乏可靠的HPLC-to-FPLC转移在净化中的强大策略.
研究的目的:
- 系统地评估染色学参数,以优化净化.
- 开发一种方法,准确地将分析HPLC净化结果转移到准备性FPLC.
- 建立一个实用和可持续的工作流程,用于净化方法的转移.
主要方法:
- 利用定义的杂物库来评估色谱参数.
- 量化了梯度度,流速,温度和移动相修饰器对分离的影响.
- 开发了一种校正方程式,以考虑化百分比的系统依赖偏差.
主要成果:
- 流速和移动相修饰剂的选择显著影响分离质量.
- 一个开发的校正方程式将化百分比的转移误差从~17%降低到<5%.
- 优化工作流程在准备性净化中实现了>90%的纯度和>30%的产量.
- 酸证明了作为三酸的更绿色替代品的潜力,改善了分离.
结论:
- 优化的染色学参数和校正方程促进了可靠的HPLC-to-FPLC方法转移用于净化.
- 既定的工作流程提高了准备性净化中的纯度和产量.
- 探索更绿色的移动相修饰剂,如酸,可以在分离中提供可持续的改进.
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