title: Opalescence in monoclonal antibody solutions and its correlation with intermolecular interactions in dilute and concentrated solutions.
abstract: Opalescence indicates physical instability of a formulation because of the presence of aggregates or liquid-liquid phase separation in solution and has been reported for monoclonal antibody (mAb) formulations. Increased solution opalescence can be attributed to attractive protein-protein interactions (PPIs). Techniques including light scattering, AUC, or membrane osmometry are routinely employed to measure PPIs in dilute solutions, whereas opalescence is seen at relatively higher concentrations, where both long- and short-range forces contribute to overall PPIs. The mAb molecule studied here shows a unique property of high opalescence because of liquid-liquid phase separation. In this study, opalescence measurements are correlated to PPIs measured in diluted and concentrated solutions using light scattering (kD ) and high-frequency rheology (G'), respectively. Charges on the molecules were calculated using zeta potential measurements. Results indicate that high opalescence and phase separation are a result of the attractive interactions in solution; however, the presence of attractive interactions do not always imply phase separation. Temperature dependence of opalescence suggests that thermodynamic contribution to opalescence is significant and Tcloud can be utilized as a potential tool to assess attractive interactions in solution.
authors.affiliation: Department of Pharmaceutical Sciences, University of Connecticut, Storrs, Connecticut, 06269.
authors.forename: Ashlesha S
authors.initials: AS
authors.lastname: Raut
authors.forename: Devendra S
authors.initials: DS
authors.lastname: Kalonia
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002626:Chemistry, Pharmaceutical; D004355:Drug Stability; D000068897:Iridescence; D008027:Light; D008956:Models, Chemical; D009994:Osmolar Concentration; D019999:Pharmaceutical Solutions; D011489:Protein Denaturation; D055550:Protein Stability; D012212:Rheology; D012542:Scattering, Radiation; D012995:Solubility; D013678:Technology, Pharmaceutical / Q000379:methods; D013696:Temperature
chemical_list: D000911:Antibodies, Monoclonal; D019999:Pharmaceutical Solutions
keywords: Protein; light scattering; liquid-liquid phase separation; monoclonal antibody; opalescence; physical characterization; protein formulation; protein-protein interactions; rheology; stability