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Scientific Paper

Measuring low levels of protein aggregation by sedimentation velocity

Reference Type:Journal Article
Author:Gabrielson, John P.;Arthur, Kelly K.
Author Addresses:Analytical Sciences Department, Amgen Inc., 4000 Nelson Road, Longmont, CO 80503, USA
Title:Measuring low levels of protein aggregation by sedimentation velocity
Secondary Title:Methods
Periodical Title:Methods
Keywords:Sedimentation velocity,Analytical ultracentrifugation,Protein aggregation,Aggregate quantitation,Therapeutic proteins,Method precision
Publishing Date: 25.12.2010
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The required performance of an analytical method depends on the purpose for which it will be used. As a methodology matures, it may find new application, and the performance demands placed on the method can increase. Sedimentation velocity analytical ultracentrifugation (SV-AUC) has a long and distinguished history with important contributions to molecular biology. Now the technique is transitioning into industrial settings, and among them, SV-AUC is now used to quantify the amount of protein aggregation in biopharmaceutical protein products, often at levels less than 1% of the total protein mass. In this paper, we review recent advances to SV methodology which have been shown to improve quantitation of protein aggregation. Then we discuss the performance of the SV method in its current state, with emphasis on the precision and quantitation limit of the method, in the context of existing industrial guidance on analytical method performance targets for quantitative methods.