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Assessment of virus production and chloramphenicol acetyl transferase expression by insect cells in serum‐free and serum‐supplemented media using a temperature‐sensitive baculovirus
Author(s) -
King G.,
Kuzio J.,
Daugulis A.,
Faulkner P.,
Allen B.,
Wu J.,
Goosen M.
Publication year - 1991
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260380918
Subject(s) - titer , multiplicity of infection , virus , spodoptera , biology , virology , chloramphenicol , microbiology and biotechnology , chemistry , recombinant dna , biochemistry , antibiotics , gene
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 × 10 6 cells/mL) compared to 3.5 × 10 6 and 8.7 × 10 6 cells/mL with the Grace's and serum‐free media, respectively. In temperature shifdown experiments with a temperature‐sensitive baculo‐virus ( acts10 YM1CAT), virus titer and chloramphenicol acetyl transferase (CAT) expression were highest in the IPL −41 (5.1 × 10 7 PFU/mL and 20000 U/mL). Use of Grace's medium gave higher virus titers than the serum‐free medium (4.4 × 10 6 vs 4.1 × 10 5 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.