A rapid, in-process assessment of virus replication is disired to quickly investigate the effects of process parameters on virus infection, and to monitor consistency of process in routine manufacturing of viral vaccines. Live virus potency assays are generally based on plaque formation, cytopathic effect, or antigen production (TCID50) and can take days to weeks to complete. Interestingly, when infected with viruses, cultured cells undergo changes in cellular metabolism that can be easily measured. These phenomena appear to be common as they has been observed in a variety of virus-host systems, e.g., in insect cells infected with baculovirus, Vero cells infected with Rotavirus, MRC-5 cells infected with Hepatitis A virus, and MRC-5 cells infected with the Varicella Zoster Virus (VZV). In this article, changes in glycolytic metabolism of MRC-5 cells as a result of CVZ infection are described. Both glucose consumption and lactate production in VZV infected MRC-5 cells are significantly elevated in comparison to uninfected cells. Based on this result, a rapid, in-process assay to follow VZV infection has been developed. The relative increase in lactate production in infected cells (α) increases as the infection progresses and then plateaus as the infection peaks. This plateau correlates with time of peak virus titer and could be used as a harvest triggering parameter in a virus production process.
Xu = cell density of uninfected cells
Xi = cell density of infected cells
XT = total cell density
Li = cumulative lactate production in infected cultures
Lu = cumulative lactate production in uninfected cultures
qLi = specific lactate production of infected cells
qLu = specific lactate production of uninfected cells
k1, K2 = constants
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Price excludes VAT (USA)
Tax calculation will be finalised during checkout.
Esanu V, Crisan I, Prahoveanu E and Sahnazarov N (1978) Isoenzymes of lactate dehydrogenase, acid phosphatase, alkaline phosphatase and peroxidase in monkey kidney cell cultures inoculated with herpes virus type 1, Virologie, 29: 1117–121.
Fisher TN, Fisher EJR (1959) Effects of cortisone and Herpes Simplex Virus on Metabolic Processes, Proc Soc Expt Biol Med, 100: 780–786.
Jain D et al. (1991a). In: Production of Biologicals from Animal Cells, Spier RE Griffiths JB, Meigner B, (eds) Butterworth-Heinemann, Oxford, UK, pp 345–350.
Jain D et al. (1991b), In: Expression Systems and Processes for rDNA Products, American Chemical Society Symposium series, #447, Hatch RT, Goochee C, Moreira and Alroy Y, (eds) American Chemical Society, Washington, D. C., pp. 97–100.
Silberklang M et al. (1995), In: Baculovirus Expression Systems and Biopesticides, Shuler ML, Wood HA, Granados RR and Hammer DA, (eds) Wiley-Liss, New York, pp 205–231.
Kamen AA, Bedard C, Tom R, Perret S and Jardin B (1996) On-line monitoring of respiration in recombinant-baculovirus infected and uninfected insect cell bioreactor cultures, Biotech Bioeng, 50: 36–48.
Krah D, Schofield T and Provost P (1990) Enhancement of varicella zoster virus plaquing efficiency with an agarose overday medium, J Virol methods, 27: 319–326.
Kussow CM, Zhou W, Gryte DM and Hu W-S (1995) Monitoring of mammalian cell growth and virus production process using on-line oxygen uptake rate measurement, Enzyme and Microbial Technology, 17: 779–783.
Spier RE. (1977) Determination of the time of harvest for foot-and-mouth disease virus cultures by measurements of the supernatant concentration of lactic dehydrogenase. Biotech Bioeng 19: 929–932.
List of Symbols
Rights and permissions
About this article
Cite this article
Singhvi, R., Markusen, J.F., Ky, B. et al. Assessment of virus infection in cultured cells using metabolic monitoring. Cytotechnology 22, 79–85 (1996). https://doi.org/10.1007/BF00353926