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Knockout of a difficult‐to‐remove CHO host cell protein, lipoprotein lipase, for improved polysorbate stability in monoclonal antibody formulations
Author(s) -
Chiu Josephine,
Valente Kristin N.,
Levy Nicholas E.,
Min Lie,
Lenhoff Abraham M.,
Lee Kelvin H.
Publication year - 2017
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.26237
Subject(s) - chinese hamster ovary cell , lipoprotein lipase , chemistry , recombinant dna , lipase , monoclonal antibody , biochemistry , polysorbate , lipoprotein , downstream processing , population , enzyme , chromatography , biology , antibody , pulmonary surfactant , cholesterol , immunology , receptor , demography , sociology , gene
While the majority of host cell protein (HCP) impurities are effectively removed in typical downstream purification processes, a small population of HCPs are particularly challenging. Previous studies have identified HCPs that are challenging for a variety of reasons. Lipoprotein lipase (LPL)—a Chinese hamster ovary (CHO) HCP that functions to hydrolyze esters in triglycerides—was one of ten HCPs identified in previous studies as being susceptible to retention in downstream processing. LPL may degrade polysorbate 80 (PS‐80) and polysorbate 20 (PS‐20) in final product formulations due to the structural similarity between polysorbates and triglycerides. In this work, recombinant LPL was found to have enzymatic activity against PS‐80 and PS‐20 in a range of solution conditions that are typical of mAb formulations. LPL knockout CHO cells were created with CRISPR and TALEN technologies and resulting cell culture harvest fluid demonstrated significantly reduced polysorbate degradation without significant impact on cell viability when compared to wild‐type samples. Biotechnol. Bioeng. 2017;114: 1006–1015. © 2016 Wiley Periodicals, Inc.