Autor(es)

Ventini, Daniella Cristina ^{[1, 2]} ; Astray, Renato Mancini ^[1] ; Nobre Lemos, Marcos Alexandre ^[1] ; Calil Jorge, Soraia Attie ^[1] ; Calderon Riquelme, Camilo ^{[3, 4]} ; Torres Suazo, Claudio Alberto ^[4] ; Tonso, Aldo ^[2] ; Pereira, Carlos Augusto ^{[1, 2]}

Afiliação do(s) autor(es)

^[1] Inst Butantan, Lab Imunol Viral, BR-05503900 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Escola Politecn, Dept Engn Quim, Lab Celulas Anim, BR-05508970 Sao Paulo - Brazil

^[3] Univ Arturo Prat, Dept Ciencias Quim & Farmaceut, Iquique - Chile

^[4] Univ Fed Sao Carlos, Dept Engn Quim, Lab Tecnol Cult Celulares, BR-13565905 Sao Carlos, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Journal of Biotechnology; v. 146, n. 4, p. 169-172, APR 15 2010.
• Citações Web of Science: 16

Resumo

S2 cell populations (S2AcRVGP2K and S2MtRVGP-Hy) were selected after transfection of gene expression vectors carrying the cDNA encoding the rabies virus glycoprotein (RVGP) gene under the control of the constitutive (actin) or inductive (metallothionein) promoters. These cell populations were cultivated in a 1 L bioreactor mimicking a large scale bioprocess. Cell cultures were carried out at 90 rpm and monitored/controlled for temperature (28 degrees C) and dissolved oxygen (10 or 50% air saturation). Cell growth attained similar to 1.5-3 x 10(7) cells/mL after 3-4 clays of cultivation. The constitutive synthesis of RVGP in S2AcRVGP2K cells led to values of 0.76 mu g/10(7) cells at day 4 of culture. The RVGP synthesis in S2MtRVGP-Hy cell fraction increased upon CuSO(4) induction attaining specific productivities of 1.5-2 mu g/10(7) cells at clays 4-5. RVGP values in supernatant as a result of cell lysis were always very low (<0.2 mu g/mL) indicating good integrity of cells in culture. Overall the RVGP productivity was of 1.5-3 mg/L. Our data showed an important influence of dissolved oxygen on RVGP synthesis allowing a higher and sustained productivity by S2MtRVGP-Hy cells when cultivated with a DO of 10% air saturation. The RVGP productivity in bioreactors shown here mirrors those previously observed for T-flasks and shaker bottles and allow the preparation of the large RVGP quantities required for studies of structure and function. (C) 2010 Elsevier B.V. All rights reserved. (AU)

• Processo FAPESP: 05/51746-6 - Expressao da proteina viral (gvp - raiva) ou da proteina fluorescente (egfp) em celulas de drosophila melanogaster.
• Beneficiário: Marcos Alexandre Nobre Lemos
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 07/52264-0 - Desenvolvimento de bioprocessos com células CHO e S2 em biorreatores para produção em larga escala de proteínas recombinantes
• Beneficiário: Daniella Cristina Ventini Monteiro
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 02/09482-3 - Expressão de genes heterólogos em células de dípteros: biologia molecular e engenharia de processos
• Beneficiário: Carlos Augusto Pereira
• Modalidade de apoio: Auxílio à Pesquisa - Temático