DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein
Autor(a) principal: | |
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Data de Publicação: | 2018 |
Outros Autores: | , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/s00253-018-8962-0 http://hdl.handle.net/11449/164193 |
Resumo: | The transmembrane rabies virus glycoprotein (RVGP) is the main antigen of vaccine formulations used around the world to prevent rabies, the most lethal preventable infectious disease known. The objective of this work was to evaluate the potential of a bioreactor using wave-induced agitation in the initial steps of scaling up the rRVGP production process by a Drosophila melanogaster S2 cell line to produce rRVGP in sufficient quantities for immunization and characterization studies. Taking advantage of some remarkable features recognized in Drosophila S2 cells for scaling the culture process, a robust recombinant lineage (S2MtRVGPH-His) engineered by our group for the expression of rRVGP using a copper-inducible promoter was used in the bioreactor cultures. The WAVE Bioreactor was chosen because it represents an innovative approach to the cultivation of animal cells using single-use technology. For that purpose, we firstly established a procedure for culturing the S2MtRVGPH-His lineage in 100 mL Schott flasks. Using an inoculum of 5 x 10(5) cells/mL in culture medium (Sf900-III) induced with solution of CuSO4 (0.7 mM) and a convenient pH range (6.2-7.0), optimal parameter values such as time of induction (72 h) and temperature (28 degrees C) to increase rRVGP production could be defined. This procedure was reproduced in culture experiments conducted in a WAVE Bioreactor (TM) 2/10 using a 2 L Cellbag. The results in Schott flasks and inWAVE Bioreactor (TM) were very similar, yielding a maximum titer of rRVGP above of 1 mg.L-1. The immunization study showed that the rRVGP produced in the bioreactor was of high immunogenic quality. |
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Repositório Institucional da UNESP |
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DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoproteinRabies virus glycoproteinRabies vaccineDrosophila melanogaster S2WAVE BioreactorScale-upRecombinant protein productionThe transmembrane rabies virus glycoprotein (RVGP) is the main antigen of vaccine formulations used around the world to prevent rabies, the most lethal preventable infectious disease known. The objective of this work was to evaluate the potential of a bioreactor using wave-induced agitation in the initial steps of scaling up the rRVGP production process by a Drosophila melanogaster S2 cell line to produce rRVGP in sufficient quantities for immunization and characterization studies. Taking advantage of some remarkable features recognized in Drosophila S2 cells for scaling the culture process, a robust recombinant lineage (S2MtRVGPH-His) engineered by our group for the expression of rRVGP using a copper-inducible promoter was used in the bioreactor cultures. The WAVE Bioreactor was chosen because it represents an innovative approach to the cultivation of animal cells using single-use technology. For that purpose, we firstly established a procedure for culturing the S2MtRVGPH-His lineage in 100 mL Schott flasks. Using an inoculum of 5 x 10(5) cells/mL in culture medium (Sf900-III) induced with solution of CuSO4 (0.7 mM) and a convenient pH range (6.2-7.0), optimal parameter values such as time of induction (72 h) and temperature (28 degrees C) to increase rRVGP production could be defined. This procedure was reproduced in culture experiments conducted in a WAVE Bioreactor (TM) 2/10 using a 2 L Cellbag. The results in Schott flasks and inWAVE Bioreactor (TM) were very similar, yielding a maximum titer of rRVGP above of 1 mg.L-1. The immunization study showed that the rRVGP produced in the bioreactor was of high immunogenic quality.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Univ Fed Sao Carlos, Dept Chem Engn, BR-13565905 Sao Carlos, SP, BrazilButantan Inst, Lab Viral Immunol, BR-05503900 Sao Paulo, SP, BrazilSao Paulo State Univ, Dept Vet Hyg & Publ Hlth, BR-18618970 Botucatu, SP, BrazilSao Paulo State Univ, Dept Vet Hyg & Publ Hlth, BR-18618970 Botucatu, SP, BrazilCNPq: 402439/2013-9SpringerUniversidade Federal de São Carlos (UFSCar)Butantan InstUniversidade Estadual Paulista (Unesp)Decarli, Monize CaiadoSantos, Diogo Peres dosAstray, Renato ManciniVentini-Monteiro, Daniella CristinaCalil Jorge, Soraia AttieCorreia, Daniela MatildeSilva, Juliana de Sa daRocca, Mayra Pereira [UNESP]Langoni, Helio [UNESP]Menozzi, Benedito Donizete [UNESP]Pereira, Carlos AugustoTorres Suazo, Claudio Alberto2018-11-26T17:51:38Z2018-11-26T17:51:38Z2018-06-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article4773-4783application/pdfhttp://dx.doi.org/10.1007/s00253-018-8962-0Applied Microbiology And Biotechnology. New York: Springer, v. 102, n. 11, p. 4773-4783, 2018.0175-7598http://hdl.handle.net/11449/16419310.1007/s00253-018-8962-0WOS:000432284700014WOS000432284700014.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengApplied Microbiology And Biotechnology1,182info:eu-repo/semantics/openAccess2023-12-11T06:19:38Zoai:repositorio.unesp.br:11449/164193Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T20:05:04.132625Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein |
title |
DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein |
spellingShingle |
DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein Decarli, Monize Caiado Rabies virus glycoprotein Rabies vaccine Drosophila melanogaster S2 WAVE Bioreactor Scale-up Recombinant protein production |
title_short |
DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein |
title_full |
DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein |
title_fullStr |
DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein |
title_full_unstemmed |
DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein |
title_sort |
DROSOPHILA S2 cell culture in a WAVE Bioreactor: potential for scaling up the production of the recombinant rabies virus glycoprotein |
author |
Decarli, Monize Caiado |
author_facet |
Decarli, Monize Caiado Santos, Diogo Peres dos Astray, Renato Mancini Ventini-Monteiro, Daniella Cristina Calil Jorge, Soraia Attie Correia, Daniela Matilde Silva, Juliana de Sa da Rocca, Mayra Pereira [UNESP] Langoni, Helio [UNESP] Menozzi, Benedito Donizete [UNESP] Pereira, Carlos Augusto Torres Suazo, Claudio Alberto |
author_role |
author |
author2 |
Santos, Diogo Peres dos Astray, Renato Mancini Ventini-Monteiro, Daniella Cristina Calil Jorge, Soraia Attie Correia, Daniela Matilde Silva, Juliana de Sa da Rocca, Mayra Pereira [UNESP] Langoni, Helio [UNESP] Menozzi, Benedito Donizete [UNESP] Pereira, Carlos Augusto Torres Suazo, Claudio Alberto |
author2_role |
author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Federal de São Carlos (UFSCar) Butantan Inst Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Decarli, Monize Caiado Santos, Diogo Peres dos Astray, Renato Mancini Ventini-Monteiro, Daniella Cristina Calil Jorge, Soraia Attie Correia, Daniela Matilde Silva, Juliana de Sa da Rocca, Mayra Pereira [UNESP] Langoni, Helio [UNESP] Menozzi, Benedito Donizete [UNESP] Pereira, Carlos Augusto Torres Suazo, Claudio Alberto |
dc.subject.por.fl_str_mv |
Rabies virus glycoprotein Rabies vaccine Drosophila melanogaster S2 WAVE Bioreactor Scale-up Recombinant protein production |
topic |
Rabies virus glycoprotein Rabies vaccine Drosophila melanogaster S2 WAVE Bioreactor Scale-up Recombinant protein production |
description |
The transmembrane rabies virus glycoprotein (RVGP) is the main antigen of vaccine formulations used around the world to prevent rabies, the most lethal preventable infectious disease known. The objective of this work was to evaluate the potential of a bioreactor using wave-induced agitation in the initial steps of scaling up the rRVGP production process by a Drosophila melanogaster S2 cell line to produce rRVGP in sufficient quantities for immunization and characterization studies. Taking advantage of some remarkable features recognized in Drosophila S2 cells for scaling the culture process, a robust recombinant lineage (S2MtRVGPH-His) engineered by our group for the expression of rRVGP using a copper-inducible promoter was used in the bioreactor cultures. The WAVE Bioreactor was chosen because it represents an innovative approach to the cultivation of animal cells using single-use technology. For that purpose, we firstly established a procedure for culturing the S2MtRVGPH-His lineage in 100 mL Schott flasks. Using an inoculum of 5 x 10(5) cells/mL in culture medium (Sf900-III) induced with solution of CuSO4 (0.7 mM) and a convenient pH range (6.2-7.0), optimal parameter values such as time of induction (72 h) and temperature (28 degrees C) to increase rRVGP production could be defined. This procedure was reproduced in culture experiments conducted in a WAVE Bioreactor (TM) 2/10 using a 2 L Cellbag. The results in Schott flasks and inWAVE Bioreactor (TM) were very similar, yielding a maximum titer of rRVGP above of 1 mg.L-1. The immunization study showed that the rRVGP produced in the bioreactor was of high immunogenic quality. |
publishDate |
2018 |
dc.date.none.fl_str_mv |
2018-11-26T17:51:38Z 2018-11-26T17:51:38Z 2018-06-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1007/s00253-018-8962-0 Applied Microbiology And Biotechnology. New York: Springer, v. 102, n. 11, p. 4773-4783, 2018. 0175-7598 http://hdl.handle.net/11449/164193 10.1007/s00253-018-8962-0 WOS:000432284700014 WOS000432284700014.pdf |
url |
http://dx.doi.org/10.1007/s00253-018-8962-0 http://hdl.handle.net/11449/164193 |
identifier_str_mv |
Applied Microbiology And Biotechnology. New York: Springer, v. 102, n. 11, p. 4773-4783, 2018. 0175-7598 10.1007/s00253-018-8962-0 WOS:000432284700014 WOS000432284700014.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Applied Microbiology And Biotechnology 1,182 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
4773-4783 application/pdf |
dc.publisher.none.fl_str_mv |
Springer |
publisher.none.fl_str_mv |
Springer |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808129158388121600 |