Analytical Ultracentrifugation and Its Role in Development and Research of Therapeutical Proteins

Methods Enzymol. 2015:562:441-76. doi: 10.1016/bs.mie.2015.04.008. Epub 2015 Aug 3.

Abstract

The historical contributions of analytical ultracentrifugation (AUC) to modern biology and biotechnology drug development and research are discussed. AUC developed by Svedberg was used to show that proteins are actually large defined molecular entities and also provided the first experimental verification for the semiconservative replication model for DNA initially proposed by Watson and Crick. This chapter reviews the use of AUC to investigate molecular weight of recombinant-DNA-produced proteins, complex formation of antibodies, intermolecular interactions in dilute and high concentration protein solution, and their impact on physical properties such as solution viscosity. Recent studies using a "competitive binding" analysis by AUC have been useful in critically evaluating the design and interpretation of surface plasmon resonance measurements and are discussed. The future of this technology is also discussed including prospects for a new higher precision analytical ultracentrifuge.

Keywords: Analytical ultracentrifugation; Competitive binding; Monoclonal antibody complexes; Protein–protein interactions; Self-association; Viscosity.

Publication types

  • Review

MeSH terms

  • Algorithms
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / isolation & purification*
  • Binding, Competitive
  • Drug Discovery
  • Humans
  • Models, Molecular
  • Molecular Weight
  • Protein Binding
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Ultracentrifugation

Substances

  • Antibodies, Monoclonal
  • Recombinant Proteins