title: Surfactant Effects on Particle Generation in Antibody Formulations in Pre-filled Syringes.
abstract: Protein aggregation and particle formation have been observed when protein solutions contact hydrophobic interfaces, and it has been suggested that this undesirable phenomenon may be initiated by interfacial adsorption and subsequent gelation of the protein. The addition of surfactants, such as polysorbate 20, to protein formulations has been proposed as a way to reduce protein adsorption at silicone oil-water interfaces and mitigate the production of aggregates and particles. In an accelerated stability study, monoclonal antibody formulations containing varying concentrations of polysorbate 20 were incubated and agitated in pre-filled glass syringes (PFS), exposing the protein to silicone oil-water interfaces at the siliconized syringe walls, air-water interfaces, and agitation stress. Following agitation in siliconized syringes that contained an air bubble, lower particle concentrations were measured in the surfactant-containing antibody formulations than in surfactant-free formulations. Polysorbate 20 reduced particle formation when added at concentrations above or below the critical micelle concentration (CMC). The ability of polysorbate 20 to decrease particle generation in PFS corresponded with its ability to inhibit gelation of the adsorbed protein layer, which was assessed by measuring the interfacial diffusion of individual antibody molecules at the silicone oil-water interface using total internal reflectance fluorescence (TIRF) microscopy with single-molecule tracking.
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado-Boulder, Boulder, Colorado.
authors.forename: Alana
authors.initials: A
authors.lastname: Gerhardt
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado-Boulder, Boulder, Colorado.
authors.forename: Aaron C
authors.initials: AC
authors.lastname: Mcumber
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado-Boulder, Boulder, Colorado.
authors.forename: Bao H
authors.initials: BH
authors.lastname: Nguyen
authors.affiliation: Formulation Sciences Department, MedImmune, Gaithersburg, Maryland.
authors.forename: Rachael
authors.initials: R
authors.lastname: Lewus
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado-Boulder, Boulder, Colorado.
authors.forename: Daniel K
authors.initials: DK
authors.lastname: Schwartz
authors.affiliation: Department of Pharmaceutical Sciences, University of Colorado-Denver, Aurora, Colorado.
authors.forename: John F
authors.initials: JF
authors.lastname: Carpenter
authors.affiliation: Department of Chemical and Biological Engineering, University of Colorado-Boulder, Boulder, Colorado. Electronic address: theodore.randolph@colorado.edu.
authors.forename: Theodore W
authors.initials: TW
authors.lastname: Randolph
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology*; D000917:Antibody Formation / Q000276:immunology*; D002626:Chemistry, Pharmaceutical / Q000379:methods; D005782:Gels / Q000737:chemistry; D011136:Polysorbates / Q000737:chemistry; D012827:Silicone Oils / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry*; D013594:Syringes; D014867:Water / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D005782:Gels; D011136:Polysorbates; D012827:Silicone Oils; D013501:Surface-Active Agents; D014867:Water
keywords: PFS; TIRFM; adsorption; interfacial diffusion; microparticles; monoclonal antibody; protein aggregation; protein gelation; silicone oil; surfactant