Heterologus gene expression in dipteran cells: molecular biology and bioprocess engineering

• Grant number: 02/09482-3
• Support type: Research Projects - Thematic Grants
• Duration: August 01, 2003 - February 28, 2010
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal researcher: Carlos Augusto Pereira
• Grantee: Carlos Augusto Pereira
• Home Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated grant(s): 07/51026-9 - Metabolism evaluation of Drosophila melanogaster S2 cells cultivated in animal protein free media producing the G glycoprotein from rabies virus, AR.EXT

Abstract

The main goal of this project is the cloning of the rabies virus glycoprotein G and the surface antigen of the human hepatitis B virus in expression vectors, for stable tranfection of Drosophila melanogaster cells (S2 cells) and constitutive or inductive protein expression (sub-project 1), as well as the optimization of insect cell technology bioprocess for production (sub-projects 2, 3 and 4). We are going to evaluate the antigenic properties of the products (sub-project 1), optimize the culture medium (sub-project 2), develop methodology for purification (sub-project 2), select the bioreactor bioprocess (sub-project 3) and establish a mathematical model for production (sub-project 4). 80, with a closed association among biological, engineering and mathematic fields, we intend to develop an original, cost effective and efficient protocol for the production of viral antigens. (AU)

Scientific publications

(21)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SUAREZ-PATINO, SANDRA FERNANDA; MANCINI, RENATO ASTRAY; PEREIRA, CARLOS AUGUSTO; TORRES SUAZO, CLAUDIO ALBERTO; MENDONCA, RONALDO ZUCATELLI; CALIL JORGE, SORAIA ATTIE. Transient expression of rabies virus glycoprotein (RVGP) in Drosophila melanogaster Schneider 2 (S2) cells. Journal of Biotechnology, v. 192, n. A, p. 255-262, DEC 20 2014. Web of Science Citations: 5.

ROSSI, NICKELI; SILVA, BRUNA G.; ASTRAY, RENATO; SWIECH, KAMILLA; PEREIRA, CARLOS A.; SUAZO, CLAUDIO A. T. Effect of hypothermic temperatures on production of rabies virus glycoprotein by recombinant Drosophila melanogaster S2 cells cultured in suspension. Journal of Biotechnology, v. 161, n. 3, p. 328-335, OCT 31 2012. Web of Science Citations: 6.

MORAES, ANGELA M.; JORGE, SORAIA A. C.; ASTRAY, RENATO M.; SUAZO, CLAUDIO A. T.; CALDERON RIQUELME, CAMILO E.; AUGUSTO, ELISABETH F. P.; TONSO, ALDO; PAMBOUKIAN, MARILENA M.; PICCOLI, ROSANE A. M.; BARRAL, MANUEL F.; PEREIRA, CARLOS A. Drosophila melanogaster S2 cells for expression of heterologous genes: From gene cloning to bioprocess development. BIOTECHNOLOGY ADVANCES, v. 30, n. 3, p. 613-628, MAY-JUN 2012. Web of Science Citations: 33.

VENTINI, DANIELLA C.; DAMIANI, RENATA; SOUSA, ALVARO P. B.; DE OLIVEIRA, JOAO E.; PERONI, CIBELE N.; RIBELA, MARIA T. C. P.; BARTOLINI, PAOLO; TONSO, ALDO; SOARES, CARLOS R. J.; PEREIRA, CARLOS A. Improved Bioprocess with CHO-hTSH Cells on Higher Microcarrier Concentration Provides Higher Overall Biomass and Productivity for rhTSH. Applied Biochemistry and Biotechnology, v. 164, n. 4, p. 401-409, JUN 2011. Web of Science Citations: 2.

BATISTA, FABIANA R. X.; GRECO, KATIA N.; ASTRAY, RENATO M.; JORGE, SORAIA A. C.; AUGUSTO, ELISABETH F. P.; PEREIRA, CARLOS A.; MENDONCA, RONALDO Z.; MORAES, ANGELA M. Behavior of Wild-type and Transfected S2 Cells Cultured in Two Different Media. Applied Biochemistry and Biotechnology, v. 163, n. 1, p. 1-13, JAN 2011. Web of Science Citations: 5.

VENTINI, DANIELLA CRISTINA; ASTRAY, RENATO MANCINI; NOBRE LEMOS, MARCOS ALEXANDRE; CALIL JORGE, SORAIA ATTIE; CALDERON RIQUELME, CAMILO; TORRES SUAZO, CLAUDIO ALBERTO; TONSO, ALDO; PEREIRA, CARLOS AUGUSTO. Recombinant rabies virus glycoprotein synthesis in bioreactor by transfected Drosophila melanogaster S2 cells carrying a constitutive or an inducible promoter. Journal of Biotechnology, v. 146, n. 4, p. 169-172, APR 15 2010. Web of Science Citations: 16.

AUGUSTO, ELISABETH F. P.; MORAES, ANGELA M.; PICCOLI, ROSANE A. M.; BARRAL, MANUEL F.; SUAZO, CLAUDIO A. T.; TONSO, ALDO; PEREIRA, CARLOS A. Nomenclature and guideline to express the amount of a membrane protein synthesized in animal cells in view of bioprocess optimization and production monitoring. BIOLOGICALS, v. 38, n. 1, p. 105-112, JAN 2010. Web of Science Citations: 8.

NOBRE LEMOS, MARCOS ALEXANDRE; DOS SANTOS, ALEXANDRA SOUZA; ASTRAY, RENATO MANCINI; PEREIRA, CARLOS AUGUSTO; CALIL JORGE, SORAIA ATTIE. Rabies virus glycoprotein expression in Drosophila S2 cells. I: Design of expression/selection vectors, subpopulations selection and influence of sodium butyrate and culture medium on protein expression. Journal of Biotechnology, v. 143, n. 2, p. 103-110, AUG 20 2009. Web of Science Citations: 16.

MENDONCA, RONALDO Z.; GRECO, KATIA N.; MORAES, ROBERTO H. P.; ASTRAY, RENATO M.; BARRAL, M. Study of kinetic parameters for the production of recombinant rabies virus glycoprotein. Cytotechnology, v. 60, n. 1-3, p. 143-151, JUL 2009. Web of Science Citations: 4.

BATISTA, FABIANA R. X.; MORAES, ANGELA M.; BUENTEMEYER, HEINO; NOLL, THOMAS. Influence of culture conditions on recombinant Drosophila melanogaster S2 cells producing rabies virus glycoprotein cultivated in serum-free medium. BIOLOGICALS, v. 37, n. 2, p. 108-118, APR 2009. Web of Science Citations: 10.

BATISTA‚ F.R.X.; PEREIRA‚ C.A.; MENDONÇA‚ R.Z.; MORAES‚ Â.M. Formulation of a protein-free medium based on IPL-41 for the sustained growth of Drosophila melanogaster S2 cells. Cytotechnology, v. 57, n. 1, p. 11-22, 2008.

GALESI‚ A.L.L.; AGUIAR‚ M.A.; ASTRAY‚ R.M.; AUGUSTO‚ E.F.P.; MORAES‚ Â.M. Growth of recombinant Drosophila melanogaster Schneider 2 cells producing rabies virus glycoprotein in bioreactor employing serum-free medium. Cytotechnology, v. 57, n. 1, p. 73-81, 2008.

JORGE‚ S.A.C.; SANTOS‚ A.S.; SPINA‚ Â.; PEREIRA‚ C.A. Expression of the hepatitis B virus surface antigen in Drosophila S2 cells. Cytotechnology, v. 57, n. 1, p. 51-59, 2008.

MENDONÇA‚ R.Z.; GRECO‚ K.N.; SOUSA‚ A.P.B.; MORAES‚ R.H.P.; ASTRAY‚ R.M.; PEREIRA‚ C.A. Enhancing effect of a protein from Lonomia obliqua hemolymph on recombinant protein production. Cytotechnology, v. 57, n. 1, p. 83-91, 2008.

PAMBOUKIAN‚ M.M.; JORGE‚ S.A.C.; SANTOS‚ M.G.; YOKOMIZO‚ A.Y.; PEREIRA‚ C.A.; TONSO‚ A. Insect cells respiratory activity in bioreactor. Cytotechnology, v. 57, n. 1, p. 37-44, 2008.

SWIECH‚ K.; ROSSI‚ N.; ASTRAY‚ R.M.; SUAZO‚ C.A.T. Enhanced production of recombinant rabies virus glycoprotein (rRVGP) by Drosophila melanogaster S2 cells through control of culture conditions. Cytotechnology, v. 57, n. 1, p. 67-72, 2008.

SWIECH‚ K.; ROSSI‚ N.; SILVA‚ B.G.; JORGE‚ S.A.C.; ASTRAY‚ R.M.; SUAZO‚ C.A.T. Bioreactor culture of recombinant Drosophila melanogaster S2 cells: characterization of metabolic features related to cell growth and production of the rabies virus glycoprotein. Cytotechnology, v. 57, n. 1, p. 61-66, 2008.

SANTOS, MARIZA G.; JORGE, SORAIA A. C.; BRILLET, KARL; PEREIRA, CARLOS A. Improving heterologous protein expression in transfected Drosophila S2 cells as assessed by EGFP expression. Cytotechnology, v. 54, n. 1, p. 15-24, May 2007.

FABIANA R. X. BATISTA; CARLOS A. PEREIRA; RONALDO Z. MENDONÇA; ANGELA M. MORAES. Evaluation of concentrated milk whey as a supplement for SF9 Spodoptera frugiperda cells in culture. ELECTRONIC JOURNAL OF BIOTECHNOLOGY, v. 9, n. 5, p. 0-0, Out. 2006.

BATISTA‚ F.R.X.; PEREIRA‚ C.A.; MENDONÇA‚ R.Z.; MORAES‚ A.M. Enhancement of Sf9 cells and baculovirus production employing Grace’s medium supplemented with milk whey ultrafiltrate. Cytotechnology, v. 49, n. 1, p. 1-9, 2005.

RODAS‚ V.M.; MARQUES‚ F.H.; HONDA‚ M.T.; SOARES‚ D.M.; JORGE‚ S.A.C.; ANTONIAZZI‚ M.M.; MEDUGNO‚ C.; CASTRO‚ M.E.B.; RIBEIRO‚ B.M.; SOUZA‚ M.L.; OTHERS. Cell culture derived AgMNPV bioinsecticide: biological constraints and bioprocess issues. Cytotechnology, v. 48, n. 1, p. 27-39, 2005.