Pharmaceutical Research

, 35:133 | Cite as

Improving Viscosity and Stability of a Highly Concentrated Monoclonal Antibody Solution with Concentrated Proline

  • Jessica J. Hung
  • Barton J. Dear
  • Aileen K. Dinin
  • Ameya U. Borwankar
  • Sumarth K. Mehta
  • Thomas T. Truskett
  • Keith P. JohnstonEmail author
Research Paper



To explain the effects of the osmolyte proline on the protein-protein interactions (PPI), viscosity and stability of highly concentrated antibody solutions in contrast to other neutral osmolytes.


The viscosity of ~225 mg/mL mAb solutions was measured with proline, glycine and trehalose as a function of pH and co-solute concentration up to 1.3 M. The stability was assessed via turbidity as well as size exclusion chromatography after 4 weeks storage at 40°C. The PPI strength was assessed qualitatively via the high concentration diffusion rate by dynamic light scattering.


Increasing proline significantly reduced the mAb viscosity and increased the colloidal stability at pH 6, but not at pH 5 further from the mAb pI. In contrast, glycine and trehalose did not improve the viscosity nor stability. The normalized diffusion coefficient at high concentration, which is inversely proportional to the attractive PPI strength, increased with proline concentration but decreased with increasing glycine.


Proline demonstrated greater efficacy for improving mAb viscosity and stability in contrast to glycine and trehalose due to its amphipathic structure and partial charge on the pyrrolidine side chain. These properties likely allow proline to screen the attractive electrostatic and hydrophobic interactions that promote self-association and high viscosities. Binary proline-histidine formulations also demonstrated greater viscosity reduction effects than histidine alone at the same total co-solute concentration, while maintaining a lower total solution osmolarity.


antibody high-concentration proline stability viscosity 





Inherent viscosity


Centrifugal filtration


Collective diffusion coefficient


Diffusion coefficient at infinite dilution








Lyophilization dilution


Monoclonal antibody




Isoelectric point


Protein-protein interactions




Size exclusion chromatography




Acknowledgments and Disclosures

AbbVie provided financial support and the antibody used in this study. The University of Texas at Austin received research funds from AbbVie Inc. to conduct the study. This work was also supported by the Welch Foundation (F-1319, F-1696) and National Science Foundation (CBET-1247945, DGE-1110007). Any opinion, findings, and conclusions or recommendations expressed in this material are those of the authors(s) and do not necessarily reflect the views of the National Science Foundation.

Supplementary material

11095_2018_2398_MOESM1_ESM.docx (176 kb)
ESM 1 (DOCX 176 kb)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Jessica J. Hung
    • 1
  • Barton J. Dear
    • 1
  • Aileen K. Dinin
    • 1
  • Ameya U. Borwankar
    • 1
  • Sumarth K. Mehta
    • 1
  • Thomas T. Truskett
    • 1
  • Keith P. Johnston
    • 1
    Email author
  1. 1.McKetta Department of Chemical EngineeringThe University of Texas at AustinAustinUSA

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