title: Polysorbate 20 prevents the precipitation of a monoclonal antibody during shear.
abstract: Monoclonal antibodies (MAbs) are widely used as therapeutic proteins and they are frequently exposed to a high degree of stress during manufacturing or delivery. MAbs shear thin upon increasing shear rates. After undergoing multiple shear cycles, with a cone-and-plate rheometer, the solution viscosity of high concentration antibodies increases due to the formation of insoluble aggregates. These shear-induced insoluble aggregates do not form when polysorbate 20 is present in solution. We hypothesize that monoclonal antibodies form a thin protein layer at the air-water interface. MAbs at the interface expose their hydrophobic core to air leading to unfolding, multiple non-specific intermolecular interactions and, upon continuous high shear, precipitation. Surface tension analysis confirms that monoclonal antibodies are surface active and that polysorbate 20 can prevent their interaction with the air-water interface. In addition, we complement these findings with a viscometer that measures bulk viscosity without the influence of an air-liquid interfacial viscosity and find that the bulk viscosity increases slightly when Mab solutions contained polysorbate 20. These methods of analysis could be used when designing manufacturing systems in which a protein solution is subject to shear forces.
authors.affiliation: Pharmaceutical Development, Genentech Inc, 1 DNA Way, South San Francisco, 94080, USA.
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Patapoff
authors.forename: Osigwe
authors.initials: O
authors.lastname: Esue
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D011232:Chemical Precipitation; D002626:Chemistry, Pharmaceutical / Q000379:methods; D005079:Excipients / Q000737:chemistry; D006801:Humans; D055595:Mechanical Phenomena; D011136:Polysorbates / Q000737:chemistry*; D011489:Protein Denaturation; D055550:Protein Stability; D012212:Rheology; D013500:Surface Tension; D013501:Surface-Active Agents / Q000737:chemistry; D013678:Technology, Pharmaceutical / Q000379:methods; D014783:Viscosity
chemical_list: D000911:Antibodies, Monoclonal; D005079:Excipients; D011136:Polysorbates; D013501:Surface-Active Agents