Abstract
The effect of serum type and concentration on recombinant protein expression in vaccinia virus infected HeLa S3 cells was studied in both static and suspension culture. A model heterologous protein,β-galactosidase (β-gal), was used. Calf and horse sera in the range of 0.5–10%(v/v) were investigated. In static culture, the calf serum concentration did not show any significant influence on the β-gal production which was almost completed within 24h postinfection (pi). Higher horse serum concentration, on the other hand, resulted in higher β-gal concentration which continued to increase until 48 h pi. Total β-gal concentrations in 0.5% calf serum at 24 h pi and 10% horse serum at 48 h pi were 2.2±0.7 and 2.2±0.1 IU/ml, respectively. In suspension culture, both sera showed their respective effects on the β-gal production similar to those observed in static culture, indicating that the cultivation method had little influence on β-gal production. Accordingly, the use of 0.5% calf serum after virus infection in recommended for economical β-gal production.
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References
Barrett N, Mitterer A, Mundt W, Eibl J, Eibl M, Gallo RC, Moss B & Dorner F (1989) Large-scale production and purification of a vaccinia recombinant-derived HIV-1 gp160 and analysis of its immunogenicity. AIDS Res. Hum. Retroviruses 5: 159–171.
Blasco R & Moss B (1992) Role of cell-associated enveloped vacinia virus in cell-to-cell spread. J. Virol. 66: 4170–4179.
Buller RML & Palumbo GJ (1991) Poxvirus pathogenesis. Microbiol. Rev. 55: 80–122.
Chakrabarti S, Brechling K & Moss B (1985) Vaccinia virus expression vector: coexpression ofβ-galactosidase provides visual screening of recombinant virus plaques. Mol. Cell. Biol. 5: 3403–3409.
Chang A & Metz DH (1976) Further investigations on the mode of entry of vaccinia virus into cells. J. Gen. Virol. 32: 275–282.
Chillakuru RA, Ryu DDY & Yilma T (1991) Propagation of recombinant vaccinia virus in HeLa cells: adsorption kinetics and replication in batch cultures. Biotechnol. Prog. 7: 85–92.
Earl PL, Cooper N & Moss B (1991) Preparation of cell cultures and vaccinia virus stocks. In: Jassen K (ed.) Current protocols in molecular biology. Vol. 2 (pp. 16.16.1–16.16.7) John Wiley & Sons, New York.
Falkner FG, Turecek PL, MacGillivray RT, Bodemer W, Scheiflinger F, Kandels S, Mitterer A, Kistner O, Barrett N, Eibl J & Dorner F (1992) High level expression of active human prothrombin in a vaccinia virus expression system. Throm. Haemost. 68: 119–124.
Janeczko RA, Rodriguez JF & Esteban M (1987) Studies on the mechanism of entry of vaccinia virus in animal cells. Arch. Virol. 92: 135–150.
Mackett M (1991) Manipulation of vaccinia virus vectors. In: Murray EJ (ed.) Methods in molecular biology. Vol. 7, Gene transfer and expression protocols. (pp. 129–146) The Humana Press, Clifton, New Jersey.
Miner JN & Hruby DE (1990) Vaccinia virus: a versatile tool for molecular biologists. Trends Biotechnol. 8: 20–25.
Moss B (1990a) Poxviridae and their replication. In: Fields BN, Knipe DM, Chanock RM, Hirsch MS, Melnick JL, Monath TP & Roizman B (eds.) Virology. Vol. 2 (pp. 2079–2111) Raven Press, New York.
Moss B (1990b) Regulation of vaccinia virus transcription. Annu. Rev. Biochem. 59: 661–688.
Moss B (1991) Vaccinia virus: a tool for research and vaccine development. Science 252: 1662–1667.
Mukhopadhyay A, Mukhopadhyay SN & Talwar GP (1994) Physiological factors of growth and susceptibility to virus regulating Vero cells for optimum yield of vaccinia and cloned gene product (β-hCG). J. Biotech. 36: 177–182.
Payne LG & Norrby E (1978) Adsorption and penetration of enveloped and naked vaccinia virus particles. J. Virol. 27: 19–27.
Pieniazek D, Pieniazek NJ, Macejak D & Luftig RB (1990) Enteric adenovirus 41 (Tak) requires low serum for growth in human primary cells. Virology 178: 72–80.
Rice CM, Franke CA, Strauss JH & Hruby DE (1985) Expression of Sindbis virus structural proteins via recombinant vaccinia virus: synthesis, processing, and incorporation into mature Sindbis virions. J. Virol. 56: 227–239.
Shin JH, Lee GM & Kim JH (1994) Comparison of cell disruption methods for determiningβ-galactosidase activity expressed in animal cells. Biotechnol. Tech. 8: 425–430.
Smith GL, Mackett M & Moss B (1983) Infectious vaccinia virus recombinants that express hepatitis B virus surface antigen. Nature 302: 490–495.
Tartaglia J & Paoletti E (1988) Recombinant vaccinia virus vaccines. Trends Biotechnol. 6: 43–46.
Thomas G, Herbert E & Hruby DE (1986) Expression and cell type-specific processing of human preproenkephalin with a vaccinia recombinant. Science 232: 1641–1643.
Wickham TJ, Davis T, Granados RR, Shuler ML & Wood HA (1992) Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system. Biotechnol. Prog. 8: 391–396.
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Jun, S.C., Lee, G.M., Chang, S.H. et al. Effect of serum type and concentration on the expression ofβ-galactosidase in recombinant vaccinia virus infected HeLa S3 cells. Cytotechnology 19, 153–159 (1995). https://doi.org/10.1007/BF00749770
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DOI: https://doi.org/10.1007/BF00749770