A critical review of analytical ultracentrifugation and field flow fractionation methods for measuring protein aggregation
A critical review of analytical ultracentrifugation and field flow fractionation methods for measuring protein aggregation
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DOI：
10.1208/aapsj080367
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发表时间：
2006-01-01
期刊：
影响因子：
4.5
通讯作者：
Shire, Steven J.
中科院分区：
文献类型：
--
作者：
Liu, Jun;Andya, James D.;Shire, Steven J.
分析超滤（AUC）和场流分级（FFF）是测量蛋白质聚集体的两种重要的生物物理方法。这两种方法都可以在广泛的溶液条件下将蛋白质单体与其聚集体形式分离。仪器和数据分析的最新进展，特别是在分析超离心技术领域，显着提高了这些生物物理方法检测蛋白质聚集体的能力和灵敏度。这些进展导致这些方法在生物制药工业中用于表征治疗性蛋白质的使用增加。然而，尽管它们比传统方法有许多优点，但仪器使用的困难和数据分析过程的复杂性往往阻碍了数据的广泛使用和正确解释。本文综述了这两种技术的最新进展，并给出了一些案例研究，以讨论它们的优点和局限性。
Analytical ultracentrifugation (AUC) and field flow fractionation (FFF) are 2 important biophysical methods for measuring protein aggregates. Both methods can separate protein monomer from its aggregate forms under a broad range of solution conditions. Recent advances in instrumentation and data analysis, particularly in the field of analytical ultracentrifugation technology, have significantly improved the capability and sensitivity of these biophysical methods for detecting protein aggregates. These advances have resulted in an increased use of these methods in the biopharmaceutical industry for characterization of therapeutic proteins. However, despite their many advantages over conventional methods, the difficulty in the use of the instrumentation and the complexity of data analysis process, have often hampered the widespread use and proper interpretation of data. This article reviews the recent progress in both technologies, and a few case studies are also presented to discuss their advantages and limitations.