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Review
. 2006 Sep 22;8(3):E590-605.
doi: 10.1208/aapsj080368.

Role of analytical ultracentrifugation in assessing the aggregation of protein biopharmaceuticals

Steven A Berkowitz  1
Affiliations
Review

Role of analytical ultracentrifugation in assessing the aggregation of protein biopharmaceuticals

Steven A Berkowitz. AAPS J. .

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 verify SEC aggregation information and the associating properties of biopharmaceuticals.

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References

    1. Kamberi M, Chung P, DeVas R, et al. Analysis of non-covalent aggregation of synthetic hPTH (1–34) by size-exclusion chromatography and the importance of suppression of non-specific interactions for precise quantitation. J Chromatogr B Analyt Technol Biomed Life Sci. 2004;810:151–155. - PubMed
    1. Clodfelter DK, Nussbaum MA, Reilly J. Comparison of free solution capillary electrophoresis and size exclusion chromatography for quantitating non-covalent aggregation of an acylated peptide. J Pharm biomed Anal. 1999;19:763–775. doi: 10.1016/S0731-7085(98)00302-1. - DOI - PubMed
    1. Link GW, Keller PL, Stout RW, Banes AJ. Effect of solution used for storage of size-exclusion columns on the subsequent chromatography of peptides and proteins. J Chromatogr. 1985;331:253–264. doi: 10.1016/0021-9673(85)80031-5. - DOI - PubMed
    1. Highley TJ, Cornell R. Supplement to LC/GC North America: The Application Notebook. Cleveland, OH: Advanstar Communications; 2004. Improve recovery and reproducibility with pretreatment of analytical TSK-gel G3000SWxl size-exclusion columns.
    1. Chirino AJ, Mire-Sluis A. Characterizing biological products and assessing comparability following manufacturing changes. Nat Biotechnol. 2004;22:1383–1391. doi: 10.1038/nbt1030. - DOI - PubMed