Type

Journal Article

Authors

Andrew Kellett
Deepak Chandran
Jonathan Bones
Sara Carillo
Michael Henry
Orla Coleman
Niall Barron
Paula Meleady
Martin Clynes
Shane McSweeney
and 1 others

Subjects

Biology

Topics
production culture media cell cell lines cell density construction materials cell growth stainless steel

Process-relevant concentrations of the leachable bDtBPP impact negatively on CHO cell production characteristics. (2016)

Abstract The biopharmaceutical industry has invested considerably in the implementation of single-use disposable bioreactors in place of or in addition to their stainless steel-counterparts. This new wave of construction materials for disposable bioprocess containers encompass a plethora of uncharacterized secondary compounds that, when in contact with the culture media, can leach, contaminating the bioprocess. One such cytotoxic leachable already receiving attention is bis(2,4-di-tert-butylphenyl)-phosphate (bDtBPP), a breakdown product of the secondary antioxidant Irgafos 168 in polyethylene-film based bags. This compound has been demonstrated to inhibit cell growth at concentrations ranging from 0.12 to 0.73 mg/L across an array of cell lines. Here we demonstrate that a further two CHO cell lines exhibit sensitivity to bDtBPP exposure at concentrations lower than that previously reported (0.035-0.1 mg/L). Furthermore, these inhibitory concentrations reflect bDtBPP levels found to leach early into the bioprocess, exposing reactor inoculums to serious risk. Quantitative label-free LC-MS/MS revealed that irrespective of cell line or concentration of bDtBPP, 8 proteins were found to be commonly differentially expressed in response to exposure to the compound highlighting biological processes related to cellular stress. Although the glycoprofile of the recombinant antibody remains primarily unchanged, we demonstrate that this compound when spiked at meaningful concentrations 72 h into culture considerably reduces the maximum cell density achieved. Studies like this reinforce the requirement for the complete characterization of all potential leachable compounds from disposable materials to assess their risk not only to the patient but also to the production pipeline itself. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 2016.
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Full list of authors on original publication

Andrew Kellett, Deepak Chandran, Jonathan Bones, Sara Carillo, Michael Henry, Orla Coleman, Niall Barron, Paula Meleady, Martin Clynes, Shane McSweeney and 1 others

Experts in our system

1
Andrew Kellett
Dublin City University
Total Publications: 29
 
2
Jonathan Bones
University College Dublin
Total Publications: 24
 
3
Michael Henry
Dublin City University
Total Publications: 78
 
4
Orla Coleman
Dublin City University
Total Publications: 6
 
5
Niall Barron
Dublin City University
Total Publications: 44
 
6
Paula Meleady
Dublin City University
Total Publications: 95
 
7
Martin Clynes
Dublin City University
Total Publications: 209