title: Polysorbate 20 Degradation in Biopharmaceutical Formulations: Quantification of Free Fatty Acids, Characterization of Particulates, and Insights into the Degradation Mechanism.
abstract: Polysorbate 20 (PS20), a commonly used surfactant in biopharmaceuticals, showed degradation upon long-term (∼18-36 months) storage of two monoclonal antibody (mAb, mAb-A, and mAb-B) drug products at 2-8 °C. The PS20 degradation resulted in the accumulation of free fatty acids (FFA), which ultimately precipitated to form particles upon long-term storage. This study documents the development, qualification, and application of a method for FFA quantification in soluble and insoluble fraction of protein formulation. The method was applied to the quantification of capric acid, lauric acid, myristic acid, palmitic/oleic acid, and stearic acid in placebo as well as active protein formulations on stability. Quantification of FFA in both the soluble and insoluble fraction of mAb-A and mAb-B provided a better mechanistic understanding of PS20 degradation and the dynamics of subsequent fatty acid particle formation. Additionally, the use of this method for monitoring and quantitation of the FFA on real time storage stability appears to aid in identifying batches with higher probability for particulate formation upon extended storage at 5 °C.
authors.affiliation: Late Stage Pharmaceutical Development and ‡Discovery Chemistry, Genentech Inc. , 1 DNA Way, South San Francisco, California 94080, United States.
authors.forename: Anthony
authors.initials: A
authors.lastname: Tomlinson
authors.affiliation: Late Stage Pharmaceutical Development and ‡Discovery Chemistry, Genentech Inc. , 1 DNA Way, South San Francisco, California 94080, United States.
authors.forename: Barthélemy
authors.initials: B
authors.lastname: Demeule
authors.affiliation: Late Stage Pharmaceutical Development and ‡Discovery Chemistry, Genentech Inc. , 1 DNA Way, South San Francisco, California 94080, United States.
authors.forename: Baiwei
authors.initials: B
authors.lastname: Lin
authors.affiliation: Late Stage Pharmaceutical Development and ‡Discovery Chemistry, Genentech Inc. , 1 DNA Way, South San Francisco, California 94080, United States.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Yadav
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001702:Biopharmaceutics / Q000379:methods*; D002626:Chemistry, Pharmaceutical; D005230:Fatty Acids, Nonesterified / Q000032:analysis*; D010316:Particle Size; D052638:Particulate Matter; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D011136:Polysorbates / Q000737:chemistry*; D012995:Solubility; D013499:Surface Properties; D013501:Surface-Active Agents / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D005230:Fatty Acids, Nonesterified; D052638:Particulate Matter; D004364:Pharmaceutical Preparations; D011136:Polysorbates; D013501:Surface-Active Agents
keywords: 1-pyrenyldiazomethane (PDAM); acid dissociation constant (pKa); biopharmaceutical formulations; evaporative light scattering detector (ELSD); free fatty acids; polysorbate 20; reverse phase ultra high performance liquid chromatography (RP-UHPLC); solid phase extraction