Contrasting the Influence of Cationic Amino Acids on the Viscosity and Stability of a Highly Concentrated Monoclonal Antibody
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To explain the effects of cationic amino acids and other co-solutes on the viscosity, stability and protein-protein interactions (PPI) of highly concentrated (≥200 mg/ml) monoclonal antibody (mAb) solutions to advance subcutaneous injection.
The viscosities of ≥200 mg/ml mAb1 solutions with various co-solutes and pH were measured by capillary rheometry in some cases up to 70,000 s−1. The viscosities are analyzed in terms of dilute PPI characterized by diffusion interaction parameters (k D ) from dynamic light scattering (DLS). MAb stability was measured by turbidity and size exclusion chromatography (SEC) after 4 weeks of 40°C storage.
Viscosity reductions were achieved by reducing the pH, or adding histidine, arginine, imidazole or camphorsulfonic acid, each of which contains a hydrophobic moiety. The addition of inorganic electrolytes or neutral osmolytes only weakly affected viscosity. Systems with reduced viscosities also tended to be Newtonian, while more viscous systems were shear thinning.
Viscosity reduction down to 20 cP at 220 mg/ml mAb1 was achieved with co-solutes that are both charged and contain a hydrophobic interaction domain for sufficient binding to the protein surface. These reductions are related to the DLS diffusion interaction parameter, k D , only after normalization to remove the effect of charge screening. Shear rate profiles demonstrate that select co-solutes reduce protein network formation.
KEY WORDSantibody arginine histidine rheology viscosity
Dynamic light scattering
Diffusion interaction parameter
Acknowledgments and Disclosures
We acknowledge support from AbbVie, the National Science Foundation (12474795) and the Welch Foundation (KPJ F-1319 and TMT F-1696). We thank Christian Reid for useful comments during the course of this work.
- 20.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;120:278–91.Google Scholar
- 38.Borwankar AU, Dear BJ, Twu A, Hung JJ, Dinin AK, Wilson BK, et al. Viscosity reduction of a concentrated monoclonal antibody with arginine·HCl and arginine·glutamate. Ind Eng Chem Res. 2016;55:11225–34.Google Scholar