Knockout of a difficult‐to‐remove CHO host cell protein, lipoprotein lipase, for improved polysorbate stability in monoclonal antibody formulations

2016/12/12 by Josephine Chiu, Kristin N. Valente, Nicholas E. Levy +3 · 27 citations
Medicine · Biochemistry, Genetics and Molecular Biology · #Monoclonal and Polyclonal Antibodies Research #Viral Infectious Diseases and Gene Expression in Insects #Protein purification and stability

paper · doi:10.1002/bit.26237

Abstract

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.

Citations

• Expression of difficult‐to‐remove host cell protein impurities during extended Chinese hamster ovary cell culture and their impact on continuous bioprocessing
• Host cell protein impurities in chromatographic polishing steps for monoclonal antibody purification

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