Effect of hypothermic temperatures on production of rabies virus glycoprotein by recombinant Drosophila melanogaster S2 cells cultured in suspension.

Autor: Rossi N; Departamento de Engenharia Química, Universidade Federal de São Carlos, Via Washington Luis, Km 235, CEP 13565-905, São Carlos-SP, Brazil. nickeli.r@hotmail.com, Silva BG, Astray R, Swiech K, Pereira CA, Suazo CA
Jazyk: angličtina
Zdroj: Journal of biotechnology [J Biotechnol] 2012 Oct 31; Vol. 161 (3), pp. 328-35. Date of Electronic Publication: 2012 Jul 20.
DOI: 10.1016/j.jbiotec.2012.05.016
Abstrakt: 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.
(Copyright © 2012 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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