Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | https://repositorio.unifesp.br/xmlui/handle/11600/63864 |
Resumo: | Monoclonal antibodies (mAbs) are molecules involved in multiple applications, ranging from use as reagents to diagnostic and therapeutic approaches. Once obtained, mAbs can be produced in vivo, by inducing ascites, as well as in vitro, by culturing the secretory hybridoma cells. In view of changes in the laws that regulate the use of animals in research, the in vivo production method has become questionable. However, the production of mAbs in vitro is not trivial. In order to replace or reduce the use of animals for this purpose, we present the present work that sought to evaluate possibilities of mAb production in vitro. For this, we use hybridomas selected based on the degree of ease in the cultivation and production of mAb, under the conditions usually practiced. Hybridomas 1F5H2 (good secretory), 5.G8 (bad secretory) and 10. D7 (best secretory) were used. The three hybridomas were studied in order to allow the comparison between in vivo and in vitro production methods regarding cell proliferation under different conditions of culture medium supplementation - 10% conventional SFB, 10% low IgG SBF and 1% Nutridoma- , as to the secretory antibody capacity of hybridomas, as well as the cost, time and volume required to produce reasonable amounts of antibodies. The cells showed no significant difference regarding the proliferative profile in the different conditions of supplementation of the culture medium. However, the 5.G8 hybridoma exhibited a higher proliferative rate when compared to the other two hybridomas, showing that the hybridoma that presented the worst performance in in vivo production, performed better in vitro and that each strain has its own characteristics. In addition, we saw that no in vitro production method, evaluated for the three hybridoma strains studied, exceeded the profitability of the in vivo method. In vitro production has always involved longer times to obtain the same amount of antibody. Supplementation of medium with Nutridoma for hybridoma culture proved to be an alternative for culture in serum-free medium. The implementation of an in vitro method alternative to the use of animals will require investments and will depend on a demand that sustains a favorable cost / benefit ratio. |
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Repositório Institucional da UNIFESP |
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Soares, Gabriela de Fátima da Silva [UNIFESP]http://lattes.cnpq.br/6404849921203695http://lattes.cnpq.br/6568232956872184Moraes, Jane Zveiter de [UNIFESP]São Paulo2022-05-24T17:06:59Z2022-05-24T17:06:59Z2020-07https://repositorio.unifesp.br/xmlui/handle/11600/63864Monoclonal antibodies (mAbs) are molecules involved in multiple applications, ranging from use as reagents to diagnostic and therapeutic approaches. Once obtained, mAbs can be produced in vivo, by inducing ascites, as well as in vitro, by culturing the secretory hybridoma cells. In view of changes in the laws that regulate the use of animals in research, the in vivo production method has become questionable. However, the production of mAbs in vitro is not trivial. In order to replace or reduce the use of animals for this purpose, we present the present work that sought to evaluate possibilities of mAb production in vitro. For this, we use hybridomas selected based on the degree of ease in the cultivation and production of mAb, under the conditions usually practiced. Hybridomas 1F5H2 (good secretory), 5.G8 (bad secretory) and 10. D7 (best secretory) were used. The three hybridomas were studied in order to allow the comparison between in vivo and in vitro production methods regarding cell proliferation under different conditions of culture medium supplementation - 10% conventional SFB, 10% low IgG SBF and 1% Nutridoma- , as to the secretory antibody capacity of hybridomas, as well as the cost, time and volume required to produce reasonable amounts of antibodies. The cells showed no significant difference regarding the proliferative profile in the different conditions of supplementation of the culture medium. However, the 5.G8 hybridoma exhibited a higher proliferative rate when compared to the other two hybridomas, showing that the hybridoma that presented the worst performance in in vivo production, performed better in vitro and that each strain has its own characteristics. In addition, we saw that no in vitro production method, evaluated for the three hybridoma strains studied, exceeded the profitability of the in vivo method. In vitro production has always involved longer times to obtain the same amount of antibody. Supplementation of medium with Nutridoma for hybridoma culture proved to be an alternative for culture in serum-free medium. The implementation of an in vitro method alternative to the use of animals will require investments and will depend on a demand that sustains a favorable cost / benefit ratio.Anticorpos monoclonais (mAbs) são moléculas envolvidas em múltiplas aplicações, que vão desde o uso como reagentes a abordagens diagnósticas e terapêuticas. Uma vez obtidos, os mAbs podem ser produzidos in vivo, por meio da indução de ascite, como também in vitro, pelo cultivo das células do hibridoma secretor. Diante de mudanças nas leis que regulam a utilização de animais em pesquisa, o método de produção in vivo passou a ser questionável. Entretanto, a produção de mAbs in vitro não é trivial. Com o objetivo de substituir ou reduzir o uso de animais para esse propósito, apresentamos o presente trabalho que buscou avaliar possibilidades de produção de mAb in vitro. Para isso, utilizamos hibridomas selecionados com base no grau de facilidade no cultivo e produção de mAb, nas condições usualmente praticadas. Foram utilizados os hibridomas 1F5H2 (bom secretor), 5.G8 (mau secretor) e 10. D7 (melhor secretor). Os três hibridomas foram estudados de modo a permitir a comparação entre os métodos de produção in vivo e in vitro quanto à proliferação celular em diferentes condições de suplementação de meio de cultura – 10% SFB convencional, 10% SFB baixa IgG e 1% Nutridoma-, quanto à capacidade secretória de anticorpos dos hibridomas, bem como custo, tempo e volume necessários para produção de quantidades razoáveis de anticorpos. As células não apresentaram diferença significativa quanto ao perfil proliferativo nas diferentes condições de suplementação do meio de cultura. Entretanto, o hibridoma 5.G8 apresentou maior taxa proliferativa quando comparado com os outros dois hibridomas, mostrando que o hibridoma que apresentava pior desempenho na produção in vivo, saiu-se melhor in vitro e que cada linhagem possui características próprias. Além disso, vimos que nenhum método de produção in vitro, avaliado para as três linhagens de hibridoma estudadas, superou a rentabilidade do método in vivo. A produção in vitro sempre envolveu tempos mais longos para a obtenção de uma mesma quantidade de anticorpo. A suplementação de meio com Nutridoma para cultura de hibridomas se mostrou uma alternativa para cultivo em meio livre de soro. A implementação de um método in vitro alternativo ao uso de animais exigirá investimentos e dependerá de uma demanda que sustente uma relação custo/benefício favorável.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)112 f.porUniversidade Federal de São PauloHibridomaCultura de célulasAnticorpo monoclonalSoro fetal bovinoNutridomaAvaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivoEvaluation of the in vitro production of monoclonal antibodies (mAbs) compared to the conventional in vivo methodinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPEscola Paulista de Medicina (EPM)Ciências Biológicas (Biologia Molecular)Avaliação da produção in vitro de anticorpos monoclonaisORIGINALBiblioteca_Gabriela.pdfBiblioteca_Gabriela.pdfDissertação de 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InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-05-16T04:27:02Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.pt_BR.fl_str_mv |
Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo |
dc.title.alternative.en.fl_str_mv |
Evaluation of the in vitro production of monoclonal antibodies (mAbs) compared to the conventional in vivo method |
title |
Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo |
spellingShingle |
Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo Soares, Gabriela de Fátima da Silva [UNIFESP] Hibridoma Cultura de células Anticorpo monoclonal Soro fetal bovino Nutridoma |
title_short |
Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo |
title_full |
Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo |
title_fullStr |
Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo |
title_full_unstemmed |
Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo |
title_sort |
Avaliação da produção in vitro de anticorpos monoclonais (mAbs) em comparação ao método convencional in vivo |
author |
Soares, Gabriela de Fátima da Silva [UNIFESP] |
author_facet |
Soares, Gabriela de Fátima da Silva [UNIFESP] |
author_role |
author |
dc.contributor.authorLattes.pt_BR.fl_str_mv |
http://lattes.cnpq.br/6404849921203695 |
dc.contributor.advisorLattes.pt_BR.fl_str_mv |
http://lattes.cnpq.br/6568232956872184 |
dc.contributor.author.fl_str_mv |
Soares, Gabriela de Fátima da Silva [UNIFESP] |
dc.contributor.advisor1.fl_str_mv |
Moraes, Jane Zveiter de [UNIFESP] |
contributor_str_mv |
Moraes, Jane Zveiter de [UNIFESP] |
dc.subject.por.fl_str_mv |
Hibridoma Cultura de células Anticorpo monoclonal Soro fetal bovino Nutridoma |
topic |
Hibridoma Cultura de células Anticorpo monoclonal Soro fetal bovino Nutridoma |
description |
Monoclonal antibodies (mAbs) are molecules involved in multiple applications, ranging from use as reagents to diagnostic and therapeutic approaches. Once obtained, mAbs can be produced in vivo, by inducing ascites, as well as in vitro, by culturing the secretory hybridoma cells. In view of changes in the laws that regulate the use of animals in research, the in vivo production method has become questionable. However, the production of mAbs in vitro is not trivial. In order to replace or reduce the use of animals for this purpose, we present the present work that sought to evaluate possibilities of mAb production in vitro. For this, we use hybridomas selected based on the degree of ease in the cultivation and production of mAb, under the conditions usually practiced. Hybridomas 1F5H2 (good secretory), 5.G8 (bad secretory) and 10. D7 (best secretory) were used. The three hybridomas were studied in order to allow the comparison between in vivo and in vitro production methods regarding cell proliferation under different conditions of culture medium supplementation - 10% conventional SFB, 10% low IgG SBF and 1% Nutridoma- , as to the secretory antibody capacity of hybridomas, as well as the cost, time and volume required to produce reasonable amounts of antibodies. The cells showed no significant difference regarding the proliferative profile in the different conditions of supplementation of the culture medium. However, the 5.G8 hybridoma exhibited a higher proliferative rate when compared to the other two hybridomas, showing that the hybridoma that presented the worst performance in in vivo production, performed better in vitro and that each strain has its own characteristics. In addition, we saw that no in vitro production method, evaluated for the three hybridoma strains studied, exceeded the profitability of the in vivo method. In vitro production has always involved longer times to obtain the same amount of antibody. Supplementation of medium with Nutridoma for hybridoma culture proved to be an alternative for culture in serum-free medium. The implementation of an in vitro method alternative to the use of animals will require investments and will depend on a demand that sustains a favorable cost / benefit ratio. |
publishDate |
2020 |
dc.date.issued.fl_str_mv |
2020-07 |
dc.date.accessioned.fl_str_mv |
2022-05-24T17:06:59Z |
dc.date.available.fl_str_mv |
2022-05-24T17:06:59Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://repositorio.unifesp.br/xmlui/handle/11600/63864 |
url |
https://repositorio.unifesp.br/xmlui/handle/11600/63864 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
112 f. |
dc.coverage.spatial.pt_BR.fl_str_mv |
São Paulo |
dc.publisher.none.fl_str_mv |
Universidade Federal de São Paulo |
publisher.none.fl_str_mv |
Universidade Federal de São Paulo |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
bitstream.url.fl_str_mv |
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bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
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