Impacts on product quality attributes of monoclonal antibodies produced in CHO cell bioreactor cultures during intentional mycoplasma contamination events

Autor: Sai Rashmika Velugula-Yellela, David N. Powers, Ryan J. Graham, Nicholas Trunfio, Erica J. Fratz‐Berilla, Adil Mohammad, Cyrus Agarabi, Talia Faison, Casey Kohnhorst, Phillip Angart
Rok vydání: 2020
Předmět:
0106 biological sciences
0301 basic medicine
biomanufacturing
medicine.drug_class
Chinese hamster ovary (CHO) cell culture
Statistics as Topic
Cell Culture Techniques
Bioengineering
CHO Cells
Monoclonal antibody
medicine.disease_cause
01 natural sciences
Applied Microbiology and Biotechnology
Article
Microbiology
Engineering Science of Biological Systems
ARTICLES
03 medical and health sciences
Bioreactors
Cricetulus
Mycoplasma
010608 biotechnology
Cricetinae
Bioreactor
medicine
Animals
principal component analysis (PCA)
Biological Products
biology
partial least squares (PLS) regression
Chemistry
bioprocessing
Chinese hamster ovary cell
Antibodies
Monoclonal
Contamination
critical quality attribute (CQA)
030104 developmental biology
Cell culture
biology.protein
monoclonal antibody (mAb)
Antibody
Critical quality attributes
Drug Contamination
Biotechnology
Zdroj: Biotechnology and Bioengineering
ISSN: 1097-0290
Popis: 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.
A mycoplasma contamination event in a biomanufacturing facility can result in costly cleanups and potential drug shortages. The authors investigated the effects of mycoplasma presence on the critical quality attributes of a monoclonal IgG1 produced in CHO cells and found that contamination increased acidic charge variants, increased DNA contamination, and led to very high percentages of high mannose species with 6 or more mannose residues.
Databáze: OpenAIRE
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