A nanolitre method to determine the hydrodynamic radius of proteins and small molecules by Taylor dispersion analysis.
A nanolitre method to determine the hydrodynamic radius of proteins and small molecules by Taylor dispersion analysis.
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DOI：
10.1016/j.ijpharm.2011.03.040
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发表时间：
2011-06
影响因子：
5.8
通讯作者：
Wendy L. Hulse;R. Forbes
中科院分区：
文献类型：
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作者：
Wendy L. Hulse;R. Forbes
正在开发的新型治疗性生物药物的数量不断增加及其高价值增加了对开发新型分析技术的需求。更快的分析时间、高精度、低样品消耗和监控工艺流程的能力都是必不可少的先决条件。我们评估了一种新的分析仪器，结合紫外区域成像和泰勒色散分析（TDA），以确定溶液中的蛋白质和小分子的流体动力学半径。对动态光散射的结果进行基准测试，我们报告了进样系统，进样量，流速，分析物浓度的影响，并强调了洗涤程序的重要性。通过将自动进样器纳入beta系统，消除了alpha实验室系统中手动进样阀导致高标准差的问题。所获得的流体动力学半径与文献值具有良好的相关性，在大多数情况下，相对标准偏差小于5%。该系统在与CE耦合后完全自动化，允许多次进样和样品/缓冲液更换，无需操作员干预。小样本量（约60 nL）、无需样品制备以及分析速度（约2-3 Mins）使得该仪器高度适用于固有不稳定、高成本的生物制药材料的实时分析，其中了解它们的聚集状态和尺寸是重要的。
The escalating number of new therapeutic biopharmaceuticals being developed and their high value increases the need for the development of novel analytical technologies. Faster analysis time, high accuracy, low sample consumption and the ability to monitor process flow are all essential prerequisites. We evaluate a novel analytical instrument that combines UV area imaging and Taylor dispersion analysis (TDA) to determine the hydrodynamic radius of proteins and small molecules in solution. Benchmarking the results against dynamic light scattering, we report the influence of injection system, injection volume, flow rates, analyte concentration and highlight the importance of washing procedures. Issues arising from the manual injection valve in the alpha laboratory system that led to high standard deviations were eliminated by incorporating an automated injector in a beta system. The hydrodynamic radii obtained show good correlation with literature values and in most cases a relative standard deviation of less than 5%.The system is fully automated after coupling to the CE which allows for multiple injections and sample/buffer changes without operator intervention. The small sample size (approx. 60 nL), the lack of sample preparation required, and the speed of analysis (approx. 2–3 mins) makes this instrument highly applicable to the real-time analysis of inherently unstable, high cost biopharmaceutical materials where understanding their aggregation state and size is important.