Analytical Sciences, Amgen Inc., 4000 Nelson Road, Longmont, CO 80503, USA.
Anal Biochem. 2010 Jan 15;396(2):231-41. doi: 10.1016/j.ab.2009.09.036. Epub 2009 Sep 24.
Sedimentation velocity analytical ultracentrifugation (SV-AUC) is routinely applied in biopharmaceutical development to measure levels of protein aggregation in protein products. SV-AUC is free from many limitations intrinsic to size exclusion chromatography (SEC) such as mobile phase and column interaction effects on protein self-association. Despite these clear advantages, SV-AUC exhibits lower precision measurements than corresponding measurements by SEC. The precision of SV-AUC is influenced by numerous factors, including sample characteristics, cell alignment, centerpiece quality, and data analysis approaches. In this study, we evaluate the precision of SV-AUC in its current practice utilizing a multilaboratory, multiproduct intermediate precision study. We then explore experimental approaches to improve SV-AUC measurement precision, with emphasis on utilization of high quality centerpieces.
沉降速度分析超速离心（SV-AUC）在生物制药开发中被常规应用于测量蛋白质产品中蛋白质聚集物的水平。SV-AUC 没有尺寸排阻色谱（SEC）所固有的许多限制，例如流动相和柱相互作用对蛋白质自组装的影响。尽管具有这些明显的优势，但 SV-AUC 的精密度测量值仍低于 SEC 的相应测量值。SV-AUC 的精密度受许多因素的影响，包括样品特征、细胞对准、离心池质量和数据分析方法。在这项研究中，我们利用多实验室、多产品中间精密度研究来评估 SV-AUC 在其实际应用中的精密度。然后，我们探讨了提高 SV-AUC 测量精密度的实验方法，重点是利用高质量的离心池。
