Type

Journal Article

Authors

Jonathan Bones
Paula Meleady
Martin Clynes
Lisa Connolly
Michael Henry
Maeve Shannon
Emma Harper
Orla Coleman
Niall Barron
Antonio Alarcon Miguez
and 6 others

Subjects

Biochemistry

Topics
seahorse xf96 single use technologies leachables bdtbpp single use bioreactors stress response single cell oxidative phosphorylation productivity growth oxidative stress mitochondria chinese hamster ovary cells endocrine disruptor cell growth cho cells

Improvements in single‐use bioreactor film material composition leads to robust and reliable Chinese hamster ovary cell performance (2019)

Abstract Single‐use 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(2,4‐di‐tert‐butylphenyl)‐phosphate (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, CHO‐K1, and CHO‐DP12, cultured in media conditioned in an older single‐use bioreactor (SUB) film (F‐1) and a newer generation film (F‐2) from the same vendor. CHO cells cultured in media conditioned for 7 days in the F‐1 film demonstrated significantly reduced growth and antibody productivity profiles when compared to controls and media conditioned for the same time period in the newer F‐2 film. Proteomic profiling of CHO cells cultured in the F‐1 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 LC–MS revealed a reduction in the abundance of the analyzed leachates from F‐2 films when compared to F‐1 films including bDtBPP, potentially explaining improved CHO cell growth in F‐2 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.
Collections Ireland -> University of Limerick -> Synthesis and Solid State Pharmaceutical Centre
Ireland -> University College Dublin -> Chemical and Bioprocess Engineering Research Collection
Ireland -> University of Limerick -> Centres Science and Engineering
Ireland -> University College Dublin -> College of Engineering & Architecture
Ireland -> University of Limerick -> Faculty of Science and Engineering
Ireland -> University College Dublin -> School of Chemical and Bioprocess Engineering

Full list of authors on original publication

Jonathan Bones, Paula Meleady, Martin Clynes, Lisa Connolly, Michael Henry, Maeve Shannon, Emma Harper, Orla Coleman, Niall Barron, Antonio Alarcon Miguez and 6 others

Experts in our system

1
Jonathan Bones
University College Dublin
Total Publications: 28
 
2
Paula Meleady
Dublin City University
Total Publications: 115
 
3
Martin Clynes
Dublin City University
Total Publications: 232
 
4
Michael Henry
Dublin City University
Total Publications: 99
 
5
Emma Harper
Dublin City University
Total Publications: 9
 
6
Orla Coleman
Dublin City University
Total Publications: 12
 
7
Niall Barron
Dublin City University
Total Publications: 60