title: Gelation of a monoclonal antibody at the silicone oil-water interface and subsequent rupture of the interfacial gel results in aggregation and particle formation.
abstract: The formation of viscoelastic gels by a monoclonal antibody (mAb) at the silicone oil-water interface was studied using interfacial shear rheology. At a concentration of 50 μg/mL, the mAb formed gels in less than 1 h, and the gelation time decreased with increasing protein concentration. To probe the effects of mechanical rupture of the interfacial gel layers, a layer of silicone oil was overlaid on the surface of aqueous solutions of mAb, and the interface was ruptured periodically with a needle. Rupture of the interfacial gel resulted in formation of subvisible particles and substantial losses of mAb monomer, which were detected by microflow imaging and quantified by size-exclusion chromatography, respectively. Resonance mass measurement showed that levels of both protein particles and silicone oil droplets increased as the gel was repeatedly ruptured with a needle. In contrast, in samples wherein the interfacial gels were not ruptured, much lower levels of aggregates and particles were detected. Addition of nonionic surfactants (polysorbate 20 or polysorbate 80) protected against aggregation and protein particle formation, with increased protection seen with increasing surfactant levels, and with the greatest inhibition observed in samples containing polysorbate 80.
authors: Department of Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, Colorado, 80045.
authors: Shyam B
authors: SB
authors: Mehta
authors: Rachael
authors: R
authors: Lewus
authors: Jared S
authors: JS
authors: Bee
authors: Theodore W
authors: TW
authors: Randolph
authors: John F
authors: JF
authors: Carpenter
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002626:Chemistry, Pharmaceutical; D002850:Chromatography, Gel; D004355:Drug Stability; D004548:Elasticity; D005782:Gels; D011136:Polysorbates / Q000737:chemistry; D066329:Protein Aggregates; D011487:Protein Conformation; D011489:Protein Denaturation; D055550:Protein Stability; D012212:Rheology; D012827:Silicone Oils / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry; D013678:Technology, Pharmaceutical / Q000379:methods; D014783:Viscosity; D014867:Water / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D005782:Gels; D011136:Polysorbates; D066329:Protein Aggregates; D012827:Silicone Oils; D013501:Surface-Active Agents; D014867:Water
keywords: Interfacial gel; formulation; monoclonal antibody; protein aggregation; rheology; surfactants