title: Effect of hypothermic temperatures on production of rabies virus glycoprotein by recombinant Drosophila melanogaster S2 cells cultured in suspension.
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.
authors: 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
authors: Nickeli
authors: N
authors: Rossi
authors: Bruna G
authors: BG
authors: Silva
authors: Renato
authors: R
authors: Astray
authors: Kamilla
authors: K
authors: Swiech
authors: Carlos A
authors: CA
authors: Pereira
authors: Claudio A T
authors: CA
authors: Suazo
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D049109:Cell Proliferation; D002478:Cells, Cultured; D004331:Drosophila melanogaster / Q000166:cytology* / Q000378:metabolism*; D006023:Glycoproteins / Q000096:biosynthesis*; D007700:Kinetics; D011820:Rabies virus / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics*; D013535:Suspensions; D013696:Temperature; D014764:Viral Proteins / Q000096:biosynthesis*
chemical_list: D006023:Glycoproteins; D013535:Suspensions; D014764:Viral Proteins