Mycoplasma are a type of bacteria that lack rigid cell walls and are occasional cell culture contaminants. In a biotechnology setting, mycoplasma could pose a threat to patient safety if undetected contaminants from the production culture remain in the final product after downstream manufacturing processes. In this study, we tracked the growth behavior of mycoplasma in co-culture with biomolecule producing CHO cells and evaluated their impact in a simulated bioreactor contamination. In addition, we investigated a new nucleic acid testing (NAT) method to detect common mycoplasma contaminant species for application in biotechnology manufacturing. Mycoplasma contamination in a bioreactor stimulates detectable changes to CHO behavior in terms of growth/viability and rates of nutrient consumption/metabolite generation approximately 48 hours after introduction. NAT detection of mycoplasma tracks consistently with established growth kinetics in early culture: at no point did the Points To Consider test detect activity where the NAT didn’t. It does not accurately reflect mycoplasma viability in late culture, as residual nucleic acids persist after the contaminants died. This argues that the NAT assay provides a better assurance of product safety in that it more sensitively detects contaminations even after they burn out.

Detection and characterization of mycoplasma contamination in cell culture with application of nucleic acid testing assays

255th National Meeting (2018)

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