Reversible self-association increases the viscosity of a concentrated monoclonal antibody in aqueous solution

J Pharm Sci. 2005 Sep;94(9):1928-40. doi: 10.1002/jps.20347.


This study was conducted to investigate the effect of reversible protein self-association on the viscosity of concentrated monoclonal antibody solutions. The viscosities of the monoclonal antibody solutions were measured by either a capillary viscometer or a cone-plate rheometer at different protein concentrations, pH, and ionic strength. Soluble aggregates were determined by size exclusion chromatography, light scattering, and analytical ultracentrifugation. Self-association of protein at high protein concentration was monitored by sedimentation equilibrium analysis using a preparative ultracentrifuge and a microfractionator. The viscosity of one of the monoclonal antibodies investigated is highly dependent on protein concentration, pH, and ionic strength of buffer and charged excipients. This antibody shows the highest viscosity near its pI at low ionic strength conditions. Sedimentation equilibrium analysis suggests that this antibody tends to reversibly self-associate at high protein concentration. The self-association appears to be quite weak and is not detectable by sedimentation velocity and size exclusion chromatography at low protein concentration. There are no significant differences in the amounts of non-dissociable soluble aggregates formed between low viscosity and high viscosity samples. These results suggest that the reversible multivalent self-association of this protein appears to be mediated mainly by electrostatic interactions of charged residues and results in unusually high viscosity of this monoclonal antibody in solution at low ionic strength conditions.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • CHO Cells
  • Chemistry, Pharmaceutical
  • Cricetinae
  • Humans
  • Hydrogen-Ion Concentration
  • Osmolar Concentration
  • Pharmaceutical Solutions / chemistry
  • Salts / chemistry
  • Sodium Chloride / chemistry
  • Solutions / chemistry*
  • Viscosity


  • Antibodies, Monoclonal
  • Pharmaceutical Solutions
  • Salts
  • Solutions
  • Sodium Chloride