Impact of Dissolved Oxygen during UV-Irradiation on the Chemical Composition and Function of CHO Cell Culture Media

Sarah M Meunier; Biljana Todorovic; Emma V Dare; Afroza Begum; Simon Guillemette; Andrew Wenger; Priyanka Saxena; J Larry Campbell; Michael Sasges; Marc G Aucoin

doi:10.1371/journal.pone.0150957

Impact of Dissolved Oxygen during UV-Irradiation on the Chemical Composition and Function of CHO Cell Culture Media

PLoS One. 2016 Mar 14;11(3):e0150957. doi: 10.1371/journal.pone.0150957. eCollection 2016.

Authors

Affiliations

¹ Waterloo Institute for Nanotechnology, Department of Chemical Engineering, University of Waterloo, Waterloo, Ontario, Canada.
² Trojan Technologies, London, Ontario, Canada.
³ SCIEX, Concord, Ontario, Canada.

Abstract

Ultraviolet (UV) irradiation is advantageous as a sterilization technique in the biopharmaceutical industry since it is capable of targeting non-enveloped viruses that are typically challenging to destroy, as well as smaller viruses that can be difficult to remove via conventional separation techniques. In this work, we investigated the influence of oxygen in the media during UV irradiation and characterized the effect on chemical composition using NMR and LC-MS, as well as the ability of the irradiated media to support cell culture. Chemically defined Chinese hamster ovary cell growth media was irradiated at high fluences in a continuous-flow UV reactor. UV-irradiation caused the depletion of pyridoxamine, pyridoxine, pyruvate, riboflavin, tryptophan, and tyrosine; and accumulation of acetate, formate, kynurenine, lumichrome, and sarcosine. Pyridoxamine was the only compound to undergo complete degradation within the fluences considered; complete depletion of pyridoxamine was observed at 200 mJ/cm2. Although in both oxygen- and nitrogen-saturated media, the cell culture performance was affected at fluences above 200 mJ/cm2, there was less of an impact on cell culture performance in the nitrogen-saturated media. Based on these results, minimization of oxygen in cell culture media prior to UV treatment is recommended to minimize the negative impact on sensitive media.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
CHO Cells
Cricetinae
Cricetulus
Culture Media / chemistry*
Oxygen / chemistry*
Ultraviolet Rays*

Substances

Culture Media
Oxygen

Grants and funding

This work was supported by the Natural Sciences and Engineering Research Council of Canada [NSERC, (www.nserccrsng.gc.ca)] ENGAGE Plus (EGP2 460753-13), and NSERC Strategic Network (MabNet, NETGP 380070-08) Grants to MGA. This work was also supported by the Natural Sciences and Engineering Research Council of Canada [NSERC, (www.nserc-crsng.gc.ca)] MITACS (www.mitacs.ca) Accelerate Post-Doctoral Fellowships to SMM, BT, EVD, AB and PS. This work was also supported by the Natural Sciences and Engineering Research Council of Canada [NSERC, (www.nserc-crsng.gc.ca), Undergraduate Student Research Assistantships to SG and AW. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. This work was also partially funded by Trojan Technologies, which also provided UV reactors and other in-kind support. This work was also supported in part by the generous contribution of Chenomx Inc. in the form of Chenomx NMR Suite 7.0-7.7 software to the Aucoin Lab. Trojan Technologies and SCIEX provided support in the form of salaries for authors MS and LC, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the 'author contributions' section.