Best Practices for Aggregate Quantitation of Antibody Therapeutics by Sedimentation Velocity Analytical Ultracentrifugation

George M. Bou-Assaf, Ivan L. Budyak, Michael Brenowitz, Eric S. Day, David Hayes, John Hill, Ranajoy Majumdar, Paola Ringhieri, Peter Schuck, Jasper C. Lin

Research output: Contribution to journalArticlepeer-review

Abstract

Analytical ultracentrifugation (AUC) is a critical analytical tool supporting the development and manufacture of protein therapeutics. AUC is routinely used as an assay orthogonal to size exclusion chromatography for aggregate quantitation. This article distills the experimental and analysis procedures used by the authors for sedimentation velocity AUC into a series of best-practices considerations. The goal of this distillation is to help harmonize aggregate quantitation approaches across the biopharmaceutical industry. We review key considerations for sample and instrument suitability, experimental design, and data analysis best practices and conversely, highlight potential pitfalls to accurate aggregate analysis. Our goal is to provide experienced users benchmarks against which they can standardize their analyses and to provide guidance for new AUC analysts that will aid them to become proficient in this fundamental technique.

Original languageEnglish (US)
Pages (from-to)2121-2133
Number of pages13
JournalJournal of Pharmaceutical Sciences
Volume111
Issue number7
DOIs
StateAccepted/In press - 2022

Keywords

  • Analytical ultracentrifugation
  • Antibody drug(s)
  • Antibody(s)
  • Biopharmaceutical characterization
  • HPLC (high performance/pressure liquid chromatography
  • Monoclonal antibody(s)
  • Protein aggregation

ASJC Scopus subject areas

  • Pharmaceutical Science

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