Assessment of virus production and chloramphenicol acetyl transferase expression by insect cells in serum‐free and serum‐supplemented media using a temperature‐sensitive baculovirus

G. King, J. Kuzio, A. Daugulis, P. Faulkner, B. Allen, Jianyong Wu, M. Goosen

Research output: Journal article publicationJournal articleAcademic researchpeer-review

17 Citations (Scopus)

Abstract

Spodoptera frugiperda insect cells were grown in Sf‐900 serum‐free medium and two kinds of serum‐supplemented media (IPL −41 and Grace's). The specific growth rates of uninfected cells were found to be 0.024, 0.35, and 0.034 h−1 respectively, at 33°C. The IPL −41 medium supported to highest maximum cell density (10.6 × 106 cells/mL) compared to 3.5 × 106 and 8.7 × 106 cells/mL with the Grace's and serum‐free media, respectively. In temperature shifdown experiments with a temperature‐sensitive baculo‐virus (acts10YM1CAT), virus titer and chloramphenicol acetyl transferase (CAT) expression were highest in the IPL −41 (5.1 × 107 PFU/mL and 20000 U/mL). Use of Grace's medium gave higher virus titers than the serum‐free medium (4.4 × 106 vs 4.1 × 105 PFU/mL) as well as higher CAT titers (7050 vs 1980 U/mL). Interestingly, in the three media used, the highest virus and CAT titers were obtained at MOI (multiplicity of infection) of 0.02 At MOI of 2.0 virtually no increase in virus of CAT titer was observed. This result is contrary to those obtained at constant‐temperature (27°C) infection and cell culture, in which higher virus titers and recombinant protein expression and obtained at higher MOI.
Original languageEnglish
Pages (from-to)1091-1099
Number of pages9
JournalBiotechnology and Bioengineering
Volume38
Issue number9
DOIs
Publication statusPublished - 1 Jan 1991
Externally publishedYes

Keywords

  • baculovirus
  • chloramphenicol acetyl transferase
  • serum‐free medium
  • Spodoptera frugiperda

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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