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Improvements in single-use bioreactor film material composition leads to robust and reliable Chinese hamster ovary cell performance
Date
2019
Abstract
Singleāuse technologies, in particular disposable bioreactor bags, have become integral within the biopharmaceutical community. However, safety concerns arose upon the identification of toxic leachable compounds derived from the plastic materials. Although the leachable bis(2,4ādiātertābutylphenyl)āphosphate (bDtBPP) has been previously shown to inhibit CHO cell growth, it is critical to determine if other compounds like this are still present in subsequent generations of films for industrial application. This study compares the performance of CHO cells, CHOāK1, and CHOāDP12, cultured in media conditioned in an older singleāuse bioreactor (SUB) film (Fā1) and a newer generation film (Fā2) from the same vendor. CHO cells cultured in media conditioned for 7 days in the Fā1 film demonstrated significantly reduced growth and antibody productivity profiles when compared to controls and media conditioned for the same time period in the newer Fā2 film. Proteomic profiling of CHO cells cultured in the Fā1 conditioned media identified differentially expressed proteins involved in oxidative stress response as well as compromised ATP synthesis. These potentially metabolically compromised cells exhibited reduced oxidative phosphorylation activity as well as lower glycolytic metabolism, characteristic of slower growing cells. Nonvolatile and metal leachables analysis of film extracts by LCāMS revealed a reduction in the abundance of the analyzed leachates from Fā2 films when compared to Fā1 films including bDtBPP, potentially explaining improved CHO cell growth in Fā2 conditioned media. Furthermore, in vitro endocrine disruptor testing of the known leachable revealed this molecule to possess the potential to act as an androgen antagonist. This study demonstrates an improvement in the materials composition used in modern generations of SUBs for safe application in the bioprocess.
Supervisor
Description
peer-reviewed
Publisher
John Wiley & Sons, Inc.
Citation
Biotechnology Progress;35 (4), e2824
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Funding code
Funding Information
Science Foundation Ireland (SFI)
Sustainable Development Goals
External Link
Type
Article
Rights
https://creativecommons.org/licenses/by-nc-sa/1.0/