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Behavior of Wild-type and Transfected S2 Cells Cultured in Two Different Media
Authors:Fabiana R. X. Batista  Kátia N. Greco  Renato M. Astray  Soraia A. C. Jorge  Elisabeth F. P. Augusto  Carlos A. Pereira  Ronaldo Z. Mendonça  Ângela M. Moraes
Affiliation:1.Departamento de Processos Biotecnológicos, Faculdade de Engenharia Química,Universidade Estadual de Campinas,Campinas,Brazil;2.Laboratório de Imunologia Viral,Instituto Butantan,S?o Paulo-SP,Brazil;3.Laboratório de Biotecnologia Industrial,Instituto de Pesquisas Tecnológicas do Estado de S?o Paulo,S?o Paulo-SP,Brazil
Abstract:An animal protein-free medium composed of IPL-41 containing 6 g L?1 yeastolate ultrafiltrate, 10 g L?1 glucose, 2 g L?1 lactose, 5 g L?1 glutamine, 1% lipid emulsion, and 0.1% Pluronic F-68 was used for producing recombinant proteins in batch mode employing two cell lines, S2AcRVGP2k expressing the G glycoprotein from rabies virus (RVGP) and S2AcHBsAgHy-9C expressing the surface antigen of hepatitis B virus (HBsAg), both obtained from Drosophila melanogaster S2 cells. Growth of wild-type S2 cells was also evaluated in the same medium. Cell behavior in the tested medium was compared to that verified in Sf900 II®. The results show that in shake flasks, S2AcRVGP2k and S2AcHBsAgHy-9C cells reached around 2?×?107 cells mL?1 in both media. In supplemented IPL-41 and Sf900 II® media, S2AcRVGP2k cells produced 367 ng RVGP mL?1 and 638 ng RVGP mL?1, respectively, while S2AcHBsAgHy-9C cells correspondently produced 573 ng HBsAg mL?1 and 322 ng HBsAg mL?1 in the mentioned media. In stirred tanks, S2AcRVGP2k cells reached 3?×?107 cells mL?1 and produced up to 758 ng RVGP mL?1. In general, glucose was consumed by cells, while lactate and ammonia were produced.
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