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Analysis Of Monoclonal Antibodies By Sedimentation Velocity Analytical Ultracentrifugation.

William B. Stine
Published 2013 · Chemistry, Medicine

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Development of a thorough understanding of the solution polydispersity of therapeutic glycoproteins including monoclonal antibodies is an important and challenging undertaking. Degradation pathways involving fragmentation could result in loss of therapeutic efficacy. Protein aggregation on the other hand is frequently considered a critical quality attribute, and concerns exist that protein aggregates could result in undesirable immunological consequences (1). Sedimentation velocity analysis performed in the analytical ultracentrifuge (SV-AUC) provides a uniquely powerful first principal measure of the hydrodynamic size and shape of proteins under conditions that can come very close to the formulated drug product. This technique avoids the potential pitfalls associated with size exclusion chromatography (SEC) including on-column dilution, adsorption or disruption of species by a stationary phase, and the need to use high ionic strength mobile phases to screen unwanted electrostatic interactions (2, 3). Furthermore, not only does SV-AUC provide a quantitative size distribution analysis, but it also provides information about macromolecular conformation. For these reasons, use of SV-AUC for analysis of therapeutic monoclonal antibodies has become widespread throughout the biopharmaceutical industry and is one of the most common orthogonal techniques to SEC for measuring aggregate and fragment levels (4-9). The studies outlined in this chapter describe the basic principles of designing, collecting, and analyzing experimental data using SV-AUC with a focus on methods for therapeutic monoclonal antibodies and other similar biologics. Details are given that facilitate the acquisition of high quality data sets that in turn simplify data analysis resulting in robust and accurate measures of solution polydispersity.
This paper references
Quantitation of aggregate levels in a recombinant humanized monoclonal antibody formulation by size-exclusion chromatography, asymmetrical flow field flow fractionation, and sedimentation velocity.
John P. Gabrielson (2007)
Is any measurement method optimal for all aggregate sizes and types?
J. Philo (2008)
Detection of protein aggregates by sedimentation velocity analytical ultracentrifugation (SV-AUC): sources of variability and their relative importance.
Kelly K. Arthur (2009)
Size-distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and lamm equation modeling.
P. Schuck (2000)
Role of analytical ultracentrifugation in assessing the aggregation of protein biopharmaceuticals
Steven A. Berkowitz (2008)
Common excipients impair detection of protein aggregates during sedimentation velocity analytical ultracentrifugation.
John P. Gabrielson (2009)
Characterizing biological products and assessing comparability following manufacturing changes
A. J. Chirino (2004)
A model for sedimentation in inhomogeneous media. I. Dynamic density gradients from sedimenting co-solutes.
P. Schuck (2004)
Quantitation of aggregates in therapeutic proteins using sedimentation velocity analytical ultracentrifugation: practical considerations that affect precision and accuracy.
A. Pekar (2007)
Analytical ultracentrifugation in the pharmaceutical industry.
J. Liu (1999)
Sedimentation velocity analytical ultracentrifugation and SEDFIT/c(s): limits of quantitation for a monoclonal antibody system.
John P. Gabrielson (2007)
Effects of protein aggregates: An immunologic perspective
A. Rosenberg (2008)

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