Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 Mar 14;11(3):e0150957.
doi: 10.1371/journal.pone.0150957. eCollection 2016.

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

Sarah M Meunier  1   2 , Biljana Todorovic  1 , Emma V Dare  1 , Afroza Begum  2 , Simon Guillemette  1 , Andrew Wenger  1 , Priyanka Saxena  2 , J Larry Campbell  3 , Michael Sasges  2 , Marc G Aucoin  1
Affiliations

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

Sarah M Meunier et al. PLoS One. .

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.

PubMed Disclaimer

Conflict of interest statement

Competing Interests: The authors of this manuscript have read the journal's policy and have the following competing interests: This work was partially funded by Trojan Technologies, which also provided UV reactors and other in-kind support. Trojan Technologies designs and manufactures equipment for UV treatment of fluids. 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. MS is employed by Trojan Technologies and JLC by SCIEX. There are no patents, products in development or marketed products to declare. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.

Figures

Fig 1
Fig 1. Experimental apparatus.
Fig 2
Fig 2. Reduction equivalent fluence (REF) based on MS2 bioassay of irradiated CD-CHO media with one UV reactor at two flow rates (50 mL/min and 100 mL/min) for both oxygen- and nitrogen-saturated media.
Fig 3
Fig 3. Concentration change with respect to UV fluence for oxygen-saturated (light gray triangles) and nitrogen-saturated (dark gray circles) media for compounds with statistically significant regression trends.
The error bars represent the standard deviation where N = 10.
Fig 4
Fig 4. Concentration change of tryptophan, kynurenine, riboflavin, and lumichrome with respect to UV fluence for oxygen-saturated (O2) and nitrogen-saturated (N2) media.
*—Significant difference between N2 and O2 saturated groups at an individual fluence level.
Fig 5
Fig 5. Concentration change of pyridoxamine with respect to UV fluence for oxygen-saturated (O2) and nitrogen-saturated (N2) media.
Fig 6
Fig 6. Comparison of the cell culture performance.
A) Time-course of viable cell density for cells grown in different fluence UV-treated nitrogen-saturated (N2) or oxygen-saturated media (O2) media. B) Total cell densities reached on Day 5 (end of exponential phase for most cultures). C) Viability of cells on Day 5. D) Total viable cell density. E) Growth rate during exponential growth phase (Days 1 through 5). The error bars represent the standard deviation for N = 6. *—Significant difference when compared to control for the same gas treatment group. #—Significant difference between N2- and O2-saturated groups at an individual fluence level.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Chen J, Bergevin J, Kiss R, Walker G, Battistoni T, Lufburrow P, et al. Case study: A novel bacterial contamination in cell culture production–Leptospira licerasiae. PDA J Pharm Sci Technol. 2012;66: 580–591. 10.5731/pdajpst.2012.00892 - DOI - PubMed
    1. Drexler HG, Uphoff CC. Mycoplasma contamination of cell cultures: Incidence, sources, effects, detection, elimination, prevention. Cytotechnology. 2002;39: 75–90. 10.1023/A:1022913015916 - DOI - PMC - PubMed
    1. Moody M, Alves W, Varghese J, Khan F. Mouse minute virus (MMV) contamination–A case study: Detection, root cause determination, and corrective actions. PDA J Pharm Sci Technol. 2011;65: 580–588. 10.5731/pdajpst.2011.00824 - DOI - PubMed
    1. Aranha H. Virus safety of biopharmaceuticals: Absence of evidence is not evidence of absence. Contract Pharma. 2011;13: 82–87.
    1. Baylis SA, Finsterbusch T, Bannert N, Blümel J, Mankertz A. Analysis of porcine circovirus type 1 detected in Rotarix vaccine. Vaccine. 2011;29: 690–697. 10.1016/j.vaccine.2010.11.028 - DOI - PubMed

Publication types

LinkOut - more resources