Detection of Protein Aggregates by Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC): Sources of Variability and Their Relative Importance
Detection of Protein Aggregates by Sedimentation Velocity Analytical Ultracentrifugation (SV-AUC): Sources of Variability and Their Relative Importance
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DOI：
10.1002/jps.21654
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发表时间：
2009-10-01
影响因子：
3.8
通讯作者：
Stoner, Michael R.
中科院分区：
文献类型：
--
作者：
Arthur, Kelly K.;Gabrielson, John P.;Stoner, Michael R.
沉降速度分析超离心（SV-AUC）已在生物制药工业中作为检测和定量蛋白质聚集体的方法得到应用。虽然该技术提供了几个优点（即，无基质分离和最少的样品处理），其结果相对于正交尺寸敏感分离技术如尺寸排阻色谱（SEC）表现出高度的可变性。这项工作的目标是表征和量化影响SV-AUC结果的变异性来源，特别是单克隆抗体单体/二聚体系统的大小分布。个别因素对整体变异性的贡献进行了研究。结果表明，样品池与旋转中心的对齐是总体变异性的最重要影响因素。其他因素的相对重要性（例如，温度平衡、时不变噪声、弯月面错位等）量化和讨论。(C)2009 Wiley-Liss，Inc.和美国药学协会药物科学杂志98：3522 - 3539，2009
Sedimentation velocity analytical ultracentrifugation (SV-AUC) has found application in the biopharmaceutical industry as a method of detecting and quantifying protein aggregates. While the technique offers several advantages (i.e., matrix-free separation and minimal sample handling), its results exhibit a high degree of variability relative to orthogonal size-sensitive separation techniques such as size exclusion chromatography (SEC). The goal of this work is to characterize and quantify the sources of variability that affect SV-AUC results, particularly size distributions for a monoclonal antibody monomer/dimer system. Contributions of individual factors to the overall variability are examined. Results demonstrate that alignment of sample cells to the center of rotation is the most significant contributing factor to overall variability. The relative importance of other factors (e.g., temperature equilibration, time-invariant noise, meniscus misplacement, etc.) are quantified and discussed. (C) 2009 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:3522-3539, 2009