Precision of protein aggregation measurements by sedimentation velocity analytical ultracentrifugation in biopharmaceutical applications.
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.
Gabrielson
John P
JP
Analytical Sciences, Amgen Inc., 4000 Nelson Road, Longmont, CO 80503, USA. jgabriel@amgen.com
Arthur
Kelly K
KK
Stoner
Michael R
MR
Winn
Bradley C
BC
Kendrick
Brent S
BS
Razinkov
Vladimir
V
Svitel
Juraj
J
Jiang
Yijia
Y
Voelker
Paul J
PJ
Fernandes
Cledwyn A
CA
Ridgeway
Ron
R
D002850:Chromatography, Gel; D011485:Protein Binding; D011487:Protein Conformation; D011506:Proteins / Q000032:analysis* / Q000737:chemistry; D014461:Ultracentrifugation / Q000379:methods*
D011506:Proteins