Improving Viscosity and Stability of a Highly Concentrated Monoclonal Antibody Solution with Concentrated Proline
- 513 Downloads
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.
Keywordsantibody high-concentration proline stability viscosity
Collective diffusion coefficient
Diffusion coefficient at infinite dilution
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.
- 14.Chang BS. Inventor; Integritybio Inc., assignee. Protein formulations containing amino acids patent WO2013063510 A1. 2013.Google Scholar
- 18.Liao SM, Du QS, Meng JZ, Pang ZW, Huang RB. The multiple roles of histidine in protein interactions. Chem Cent J. 2013;7Google Scholar
- 22.Baumann P, Schermeyer M-T, Burghardt H, Dürr C, Gärtner J, Hubbuch J. Prediction and characterization of the stability enhancing effect of the Cherry-Tag™ in highly concentrated protein solutions by complex rheological measurements and MD simulations. Int J Pharm. 2017;531(1):360–71.CrossRefPubMedGoogle Scholar
- 42.Nicoud L, Jagielski J, Pfister D, Lazzari S, Massant J, Lattuada M, et al. Kinetics of Monoclonal Antibody Aggregation from Dilute toward Concentrated Conditions. J Phys Chem B. 2016.Google Scholar
- 46.Calero-Rubio C, Ghosh R, Saluja A, Roberts CJ. Predicting protein-protein interactions of concentrated antibody solutions using dilute solution data and coarse-grained molecular models. J Pharm Sci. 2017;Google Scholar
- 47.Godfrin PD, Zarzar J, Zarraga IE, Porcar L, Falus P, Wagner NJ, et al. The effect of hierarchical cluster formation on the viscosity of concentrated monoclonal antibody formulations studied by neutron scattering. J Phys Chem B. 2015.Google Scholar
- 55.Wang S, Zhang N, Hu T, Dai WG, Feng X, Zhang X, et al. Viscosity-Lowering Effect of Amino Acids and Salts on Highly Concentrated Solutions of two IgG1 Monoclonal Antibodies. Mol Pharmaceutics. 2015.Google Scholar
- 56.Saha D, Joshi YM, Bandyopadhyay R. Characteristics of the secondary relaxation process in soft colloidal suspensions. Epl. 2015;112(4)Google Scholar