title: Residual Host Cell Protein Promotes Polysorbate 20 Degradation in a Sulfatase Drug Product Leading to Free Fatty Acid Particles.
abstract: This study investigated the root cause behind an observed free fatty acid particle formation and resulting Polysorbate 20 (PS20) loss for a sulfatase drug product upon long-term storage at 5 ± 3°C. Reversed- phase chromatography with mass spectrometric analysis as well as charged aerosol detection was used to characterize the peaks associated with the intact and degraded PS20. Additionally, a proteomics study was undertaken to identify the residual host cell proteins in the sulfatase drug substance. PS20 stability studies were conducted in the presence of sulfatase, a sulfatase inhibitor, putative phospholipase B-like 2, and mock drug substance produced using a null cell line vector under experimental conditions optimized for PS20 degradation. This study provides the first published evidence where the residual host cell protein present in the drug substance was identified and experimentally shown to catalyze the breakdown of PS20 in a protein formulation over time, resulting in free fatty acid particles and PS20 loss. This study demonstrates the importance of early detection of potential impurities in the protein drug substance that may contribute to polysorbate degradation to make a judicious selection of the surfactant and its optimized concentration for the final drug product.
authors.affiliation: Drug and Combination Product Development, Shire, Lexington, Massachusetts 02421. Electronic address: ndixit@shire.com.
authors.forename: Nitin
authors.initials: N
authors.lastname: Dixit
authors.affiliation: Drug and Combination Product Development, Shire, Lexington, Massachusetts 02421.
authors.forename: Nazila
authors.initials: N
authors.lastname: Salamat-Miller
authors.affiliation: Analytical Development, Shire, Lexington, Massachusetts 02421.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Salinas
authors.affiliation: Drug and Combination Product Development, Shire, Lexington, Massachusetts 02421.
authors.forename: Katherine D
authors.initials: KD
authors.lastname: Taylor
authors.affiliation: Drug and Combination Product Development, Shire, Lexington, Massachusetts 02421.
authors.forename: Sujit K
authors.initials: SK
authors.lastname: Basu
mesh_terms: D005230:Fatty Acids, Nonesterified / Q000032:analysis / Q000378:metabolism*; D006801:Humans; D008245:Lysophospholipase / Q000032:analysis / Q000378:metabolism*; D010316:Particle Size; D011136:Polysorbates / Q000032:analysis / Q000378:metabolism*; D055550:Protein Stability; D040901:Proteomics / Q000379:methods; D013429:Sulfatases / Q000032:analysis / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D005230:Fatty Acids, Nonesterified; D011136:Polysorbates; D008245:Lysophospholipase; D013429:Sulfatases
keywords: fatty acid particles; host cell proteins; polysorbate degradation; protein stability; putative phospholipase B-like 2; sulfatase