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

Author(s)

Rossi, Nickeli ^[1] ; Silva, Bruna G. ^[1] ; Astray, Renato ^[2] ; Swiech, Kamilla ^[1] ; Pereira, Carlos A. ^[2] ; Suazo, Claudio A. T. ^[1]

Affiliation

^[1] Univ Fed Sao Carlos, Dept Engn Quim, BR-13565905 Sao Carlos, SP - Brazil

^[2] Inst Butantan, Lab Imunol Viral, BR-05503900 Sao Paulo - Brazil

• Document type: Journal article
• Source: Journal of Biotechnology; v. 161, n. 3, p. 328-335, OCT 31 2012.
• Web of Science Citations: 6

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 degrees C. The maximum specific growth rates mu(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 degrees C) was then tested in a bioreactor with control of pH and dissolved oxygen in batch and fed-batch modes. In the batch culture, mu(max) and C-max(rRVGP) were 0.060 h(-1) and 0.149 mg L-1 at 28 degrees C and 0.026 h(-1) and 0.354 mg L-1 at 20 degrees C, respectively. One batch-culture experiment was carried out with adaptation of the cells by the temperature falling in steps from 20 degrees C to 16 degrees C, so that mu(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 degrees C, mu(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 degrees C, when mu(max) is kept low and with feeding of limitant aminoacids. (C) 2012 Elsevier B.V. All rights reserved. (AU)

• FAPESP's process: 02/09482-3 - Heterologus gene expression in dipteran cells: molecular biology and bioprocess engineering
• Grantee: Carlos Augusto Pereira
• Support Opportunities: Research Projects - Thematic Grants