Impacts on product quality attributes of monoclonal antibodies produced in CHO cell bioreactor cultures during intentional mycoplasma contamination events |
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Authors: | Erica J Fratz-Berilla Phillip Angart Ryan J Graham David N Powers Adil Mohammad Casey Kohnhorst Talia Faison Sai Rashmika Velugula-Yellela Nicholas Trunfio Cyrus Agarabi |
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Institution: | 1. Division of Biotechnology Review and Research II, U.S. Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Office of Biotechnology Products, Silver Spring, Maryland;2. Division of Product Quality Research, U.S. Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Office of Testing and Research, Silver Spring, Maryland
Department of Chemical Engineering, University of Massachusetts Lowell, Lowell, Massachusetts;3. Division of Product Quality Research, U.S. Food and Drug Administration, Center for Drug Evaluation and Research, Office of Pharmaceutical Quality, Office of Testing and Research, Silver Spring, Maryland;4. Nova Biomedical, Waltham, Massachusetts;5. Sartorius Stedim Data Analytics, Bohemia, New York |
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Abstract: | A mycoplasma contamination event in a biomanufacturing facility can result in costly cleanups and potential drug shortages. Mycoplasma may survive in mammalian cell cultures with only subtle changes to the culture and penetrate the standard 0.2-µm filters used in the clarification of harvested cell culture fluid. Previously, we reported a study regarding the ability of Mycoplasma arginini to persist in a single-use, perfusion rocking bioreactor system containing a Chinese hamster ovary (CHO) DG44 cell line expressing a model monoclonal immunoglobulin G 1 (IgG1) antibody. Our previous work showed that M. arginini affects CHO cell growth profile, viability, nutrient consumption, oxygen use, and waste production at varying timepoints after M. arginini introduction to the culture. Careful evaluation of certain identified process parameters over time may be used to indicate mycoplasma contamination in CHO cell cultures in a bioreactor before detection from a traditional method. In this report, we studied the changes in the IgG1 product quality produced by CHO cells considered to be induced by the M. arginini contamination events. We observed changes in critical quality attributes correlated with the duration of contamination, including increased acidic charge variants and high mannose species, which were further modeled using principal component analysis to explore the relationships among M. arginini contamination, CHO cell growth and metabolites, and IgG1 product quality attributes. Finally, partial least square models using NIR spectral data were used to establish predictions of high levels (≥104 colony-forming unit CFU/ml]) of M. arginini contamination, but prediction of levels below 104 CFU/ml were not reliable. Contamination of CHO cells with M. arginini resulted in significant reduction of antibody product quality, highlighting the importance of rapid microbiological testing and mycoplasma testing during particularly long upstream bioprocesses to ensure product safety and quality. |
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Keywords: | biomanufacturing bioprocessing Chinese hamster ovary (CHO) cell culture critical quality attribute (CQA) monoclonal antibody (mAb) mycoplasma partial least squares (PLS) regression principal component analysis (PCA) |
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