Effect of hypothermic temperatures on production of rabies virus glycoprotein by recombinant Drosophila melanogaster S2 cells cultured in suspension.
J Biotechnol
; 161(3): 328-35, 2012 Oct 31.
Article
em En
| MEDLINE
| ID: mdl-22820340
ABSTRACT
Aiming at maximizing the production of transmembrane rabies virus glycoprotein (rRVGP), the influence of hypothermic temperature on a recombinant Drosophila melanogaster S2 cell culture in Sf-900II medium was investigated. Cell growth and rRVGP production were assessed at 4 culture temperatures in Schott flasks 16, 20, 24 and 28 °C. The maximum specific growth rates µ(max) were, respectively 0.009, 0.019, 0.038 and 0.035 h(-1), while the maximum rRVGP levels C(max)(rRVGP) were 0.075, 2.973, 0.480 and 1.404 mg L(-1). The best production temperature (20 °C) was then tested in a bioreactor with control of pH and dissolved oxygen in batch and fed-batch modes. In the batch culture, µ(max) and C(max)(rRVGP) were 0.060 h(-1) and 0.149 mg L(-1) at 28 °C and 0.026 h(-1) and 0.354 mg L(-1) at 20 °C, respectively. One batch-culture experiment was carried out with adaptation of the cells by the temperature falling in steps from 20 °C to 16 °C, so that µ(max) fell from 0.023 to 0.013 h(-1), while C(max)(rRVGP) was improved to 0.567 mg L(-1). In the fed-batch mode at 20 °C, µ(max) was 0.025 h(-1) and C(max)(rRVGP) was 1.155 mg L(-1). Taken together, these results indicate that the best strategy for optimized rRVGP production is the culture at hypothermic temperature of 20 °C, when µ(max) is kept low and with feeding of limitant aminoacids.
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Base de dados:
MEDLINE
Assunto principal:
Vírus da Raiva
/
Recombinação Genética
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Temperatura
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Proteínas Virais
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Glicoproteínas
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Drosophila melanogaster
Idioma:
En
Ano de publicação:
2012
Tipo de documento:
Article