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. 2006 Sep 22;8(3):E590–E605. doi: 10.1208/aapsj080368

Role of analytical ultracentrifugation in assessing the aggregation of protein biopharmaceuticals

Steven A Berkowitz 1,✉
PMCID: PMC2761066  PMID: 17025277

Abstract

In developing and manufacturing protein biopharmaceuticals, aggregation is a parameter that needs careful monitoring to ensure the quality and consistency of the final biopharmaceutical drug product. The analytical method of choice used to perform this task is size-exclusion chromatography (SEC). However, it is becoming more and more apparent that considerable care is required in assessing the accuracy of SEC data. One old analytical tool that is now reappearing to help in this assessment is analytical ultracentrifugation (AUC). Developments in AUC hardware and, more importantly, recent developments in AUC data analysis computer programs have converged to provide this old biophysical tool with the ability to extract very high resolution size information about the molecules in a given sample from a simple sedimentation velocity experiment. In addition, AUC allows sample testing to be conducted in the exact or nearly exact liquid formulation or reconstituted liquid formulation of the biopharmaceutical in the vial, with minimal surface area contact with extraneous materials. As a result, AUC analysis can provide detailed information on the aggregation of a biopharmaceutical, while avoiding many of the major problems that can plague SEC, thus allowing AUC to be used as an orthogonal method to verity SEC aggregation information and the associating properties of biopharmaceuticals.

Keywords: Protein aggregation, analytical ultracentrifugation, size-exclusion chromatography, SEDFIT

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