Behavior of Monoclonal Antibodies: Relation Between the Second Virial Coefficient (B 2) at Low Concentrations and Aggregation Propensity and Viscosity at High Concentrations
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To investigate relationship between second virial coefficient B 2 and viscosity and aggregation propensity of highly concentrated monoclonal antibody (MAbs) solutions.
Intermolecular interactions of 3 MAbs solutions with varying pH were characterized according to B 2 estimated by analytical ultracentrifugation sedimentation equilibrium with initial loading concentrations <10 mg/mL. Viscosity measurements and stability assessments of MAb solutions at concentrations higher than 100 mg/mL were conducted.
B 2 of all MAb solutions depended on solution pH and have qualitative correlation with viscosity and aggregation propensity. The more negative the B 2 values, the more viscous the solution, acquiring increased propensity to aggregate. Solutions with B 2 values of ~2 × 10−5 mL·mol/g2 acquire similar viscosity and aggregation propensity regardless of amino acid sequences; for solutions with negative B 2 values, viscosity and aggregation propensity differed depending on sequences. Results suggest attractive intermolecular interactions represented by negative B 2 values are influenced by surface properties of individual MAbs.
B 2 can be used, within certain limitations, as an effective indicator of viscosity and aggregation propensity of highly concentrated MAb solutions.
KEY WORDSaggregation analytical ultracentrifugation sedimentation equilibrium monoclonal antibody second virial coefficient viscosity
ACKNOWLEDGMENTS & DISCLOSURES
The authors would like to thank Daisuke Ama, Kei Kubota, Yuki Araki, Mami Mitsui and Misako Sawakuri (Daiichi Sankyo Co., Ltd., Tokyo, Japan) for their skillful technical support.
- 23.Saluja A, Badkar AV, Zeng DL, Nema S, Kalonia DS. Ultrasonic storage modules as a novel parameter for analyzing protein-protein interactions in high protein concentration solutions: correlation with static and dynamic light scattering measurements. Biophys J. 2007;92:234–44.PubMedCrossRefGoogle Scholar
- 24.Holde KE, Johnson C, Ho PS. Principles of physical biochemistry. Upper Saddle River: Pearson Education; 2006.Google Scholar
- 29.Brun VL, Friess W, Bassarab S, Mühlau S, Garidel P. A critical evaluation of self-interaction chromatography as a predictive tool for the assessment of protein-protein interactions in protein formulation development: a case study of a therapeutic monoclonal antibody. Eur J Pharm Biopharm. 2010;75(1):16–25.PubMedCrossRefGoogle Scholar
- 31.Harding SE, Rowe AJ, Horton JC. Analytical ultracentrifugation in biochemistry and polymer science. London: Royal Society of Chemistry; 1992. p. 90–125.Google Scholar
- 37.Yamakawa H. Concentration dependence of the frictional coefficient of polymers in solution. 1962;36(11):2995–3001.Google Scholar
- 40.Deszczynski M, Harding SE, Winzor DJ. Negative second virial coefficients as predictors of protein crystal growth: evidence from sedimentation equilibrium studies that refutes the designation of those light scattering parameters as osmotic virial coefficients. Biophys Chem. 2006;120:106–13.PubMedCrossRefGoogle Scholar