Recombinant rabies virus glycoprotein synthesis in bioreactor by transfected Drosophila melanogaster S2 cells carrying a constitutive or an inducible promoter

Author(s)

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]}

Affiliation

^[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

• Document type: Journal article
• Source: Journal of Biotechnology; v. 146, n. 4, p. 169-172, APR 15 2010.
• Web of Science Citations: 16

Abstract

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)

• 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