Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae)
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/4977 |
Resumo: | The Neospora caninum is a protozoan apicomplexan with implication in abortions in many countries. They are included in the economical losses, the abortament, the proportion of discarded cows and the replacement of new animals in the flock, the fall in the milk production, as well as in the fat production in the milk. It is known that the kits existent diagnoses in the healthy market of high cost and they are not produced at the national market, it is needed, to offer diagnosis alternatives reliable, and fast, with national technology. The proposal of the present work was the production of monoclonal antibodies (mAb) of high likeness against the Neospora caninum (sample Nc-1), for the use in immunodiagnostic tests as immunofluorescence, immunohistochemistry, ELISA and dot-ELISA. For the production of mAb the sonicated protozoan was used, originating from the culture in cells VERO and it was purified by filtration, it was used the tachyzoites for the immunization of the mice BALB/c, using as saponin adjuvant , what allowed the obtaining of polyclonal antibodies. The fusing of the esplenic cells of the immunized mice and the myelomas SP2/0 resulted in the obtaining of 72,4% of hybridomas secretors of antibodies anti-Nc-1. After the cloning for limiting dilution was obtained 78,2% of the clones secretors of antibodies anti-Nc- 1. After the recloning for limiting dilution, they were obtained 4 clones secretors of mAb of the class IgG. Obtained mAb IgG2a were capable to reveal the proteins of mass molecular 25 kDa and 70 kDa for "western blotting" to inhibit the multiplication of the taquyzoites in cells VERO. The monoclonal antibodies produced can be used in the development of more sensitive tests, in the epidemic studies of the neosporosis or they can be used in the identification of relevant proteins for the development of vaccines. |
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Devens, Bruna Alveshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4711727Y4Salcedo, Joaquín Hernán Patarroyohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783313T4Benjamin, Laércio dos Anjoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797917E7Viloria, Marlene Isabel Vargashttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781964E6Guimarães, Antonio Marcoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786353E2Sales Junior, Policarpo Ademarhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4764903E9Oliveira, Leandro Licursi dehttp://lattes.cnpq.br/05782313922181622015-03-26T13:46:40Z2009-08-032015-03-26T13:46:40Z2009-02-20DEVENS, Bruna Alves. Production of hybridomas and clones to obtain monoclonal antibodies anti - Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae). 2009. 3 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2009.http://locus.ufv.br/handle/123456789/4977The Neospora caninum is a protozoan apicomplexan with implication in abortions in many countries. They are included in the economical losses, the abortament, the proportion of discarded cows and the replacement of new animals in the flock, the fall in the milk production, as well as in the fat production in the milk. It is known that the kits existent diagnoses in the healthy market of high cost and they are not produced at the national market, it is needed, to offer diagnosis alternatives reliable, and fast, with national technology. The proposal of the present work was the production of monoclonal antibodies (mAb) of high likeness against the Neospora caninum (sample Nc-1), for the use in immunodiagnostic tests as immunofluorescence, immunohistochemistry, ELISA and dot-ELISA. For the production of mAb the sonicated protozoan was used, originating from the culture in cells VERO and it was purified by filtration, it was used the tachyzoites for the immunization of the mice BALB/c, using as saponin adjuvant , what allowed the obtaining of polyclonal antibodies. The fusing of the esplenic cells of the immunized mice and the myelomas SP2/0 resulted in the obtaining of 72,4% of hybridomas secretors of antibodies anti-Nc-1. After the cloning for limiting dilution was obtained 78,2% of the clones secretors of antibodies anti-Nc- 1. After the recloning for limiting dilution, they were obtained 4 clones secretors of mAb of the class IgG. Obtained mAb IgG2a were capable to reveal the proteins of mass molecular 25 kDa and 70 kDa for "western blotting" to inhibit the multiplication of the taquyzoites in cells VERO. The monoclonal antibodies produced can be used in the development of more sensitive tests, in the epidemic studies of the neosporosis or they can be used in the identification of relevant proteins for the development of vaccines.O Neospora caninum é um protozoário apicomplexa com implicação em abortamentos em muitos países. Estão inclusas nas perdas econômicas, os abortamentos, a proporção de vacas descartadas e a reposição de novos animais no rebanho, a queda na produção leiteira, bem como na produção de gordura no leite. Sabe-se que os kits diagnósticos existentes no mercado são de alto custo e não são produzidos no mercado nacional, necessita-se, oferecer alternativas de diagnóstico confiáveis, e rápidas, com tecnologia nacional. A proposta do presente trabalho foi a produção de anticorpos monoclonais (AcM) de alta afinidade contra o Neospora caninum (amostra Nc-1), para a utilização em testes de imunodiagnóstico como imunofluorescência, imunoistoquímica, ELISA e dot-ELISA. Para a produção dos AcM usou-se o protozoário sonicado, proveniente da cultura em células VERO e foi purificado por filtração, utilizaram-se os taquizoítos para a imunização dos camundongos BALB/c, usando como adjuvante a saponina, o que permitiu a obtenção de anticorpos policlonais. A fusão das células esplênicas, provenientes dos camundongos imunizados, com as células de mieloma SP2/0 resultou na obtenção de 72,4% de hibridomas secretores de anticorpos anti-Nc-1. Após a clonagem por diluição limitante obteve-se 78,2% dos clones secretores de anticorpos anti-Nc-1. Após a reclonagem por diluição limitante, foram obtidos 4 clones secretores de AcM da classe IgG. Os AcM IgG2a obtidos foram capazes de revelar as proteínas de massa molecular 25 kDa e 70 kDa por western blotting e inibirem a multiplicação dos taquizoítos em células VERO. Os anticorpos monoclonais produzidos poderão ser usados no desenvolvimento de testes mais sensíveis para estudos epidemiológicos da neosporose ou poderão ser utilizados na identificação de proteínas relevantes para o desenvolvimento de vacinas.Fundação de Amparo a Pesquisa do Estado de Minas Geraisapplication/pdfporUniversidade Federal de ViçosaMestrado em Medicina VeterináriaUFVBRBiotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. deNeospora caninumAnticorpos monoclonaisNeospora caninumMonoclonal antibodiesCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMALProdução de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae)Production of hybridomas and clones to obtain monoclonal antibodies anti - Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINAL01 - capa_abstract.pdfapplication/pdf58716https://locus.ufv.br//bitstream/123456789/4977/1/01%20-%20capa_abstract.pdf8427f1a81ed09f080394210a61912045MD51TEXT01 - capa_abstract.pdf.txt01 - capa_abstract.pdf.txtExtracted texttext/plain5030https://locus.ufv.br//bitstream/123456789/4977/2/01%20-%20capa_abstract.pdf.txt5ea4785cd47d7998e47c772e9c51843eMD52THUMBNAIL01 - capa_abstract.pdf.jpg01 - capa_abstract.pdf.jpgIM Thumbnailimage/jpeg3618https://locus.ufv.br//bitstream/123456789/4977/3/01%20-%20capa_abstract.pdf.jpg6e91458d56b48f90b8440965ff24d6a9MD53123456789/49772016-04-11 23:06:14.731oai:locus.ufv.br:123456789/4977Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-12T02:06:14LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) |
dc.title.alternative.eng.fl_str_mv |
Production of hybridomas and clones to obtain monoclonal antibodies anti - Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) |
title |
Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) |
spellingShingle |
Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) Devens, Bruna Alves Neospora caninum Anticorpos monoclonais Neospora caninum Monoclonal antibodies CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL |
title_short |
Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) |
title_full |
Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) |
title_fullStr |
Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) |
title_full_unstemmed |
Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) |
title_sort |
Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae) |
author |
Devens, Bruna Alves |
author_facet |
Devens, Bruna Alves |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4711727Y4 |
dc.contributor.author.fl_str_mv |
Devens, Bruna Alves |
dc.contributor.advisor-co1.fl_str_mv |
Salcedo, Joaquín Hernán Patarroyo |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783313T4 |
dc.contributor.advisor-co2.fl_str_mv |
Benjamin, Laércio dos Anjos |
dc.contributor.advisor-co2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797917E7 |
dc.contributor.advisor1.fl_str_mv |
Viloria, Marlene Isabel Vargas |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4781964E6 |
dc.contributor.referee1.fl_str_mv |
Guimarães, Antonio Marcos |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786353E2 |
dc.contributor.referee2.fl_str_mv |
Sales Junior, Policarpo Ademar |
dc.contributor.referee2Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4764903E9 |
dc.contributor.referee3.fl_str_mv |
Oliveira, Leandro Licursi de |
dc.contributor.referee3Lattes.fl_str_mv |
http://lattes.cnpq.br/0578231392218162 |
contributor_str_mv |
Salcedo, Joaquín Hernán Patarroyo Benjamin, Laércio dos Anjos Viloria, Marlene Isabel Vargas Guimarães, Antonio Marcos Sales Junior, Policarpo Ademar Oliveira, Leandro Licursi de |
dc.subject.por.fl_str_mv |
Neospora caninum Anticorpos monoclonais |
topic |
Neospora caninum Anticorpos monoclonais Neospora caninum Monoclonal antibodies CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL |
dc.subject.eng.fl_str_mv |
Neospora caninum Monoclonal antibodies |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL |
description |
The Neospora caninum is a protozoan apicomplexan with implication in abortions in many countries. They are included in the economical losses, the abortament, the proportion of discarded cows and the replacement of new animals in the flock, the fall in the milk production, as well as in the fat production in the milk. It is known that the kits existent diagnoses in the healthy market of high cost and they are not produced at the national market, it is needed, to offer diagnosis alternatives reliable, and fast, with national technology. The proposal of the present work was the production of monoclonal antibodies (mAb) of high likeness against the Neospora caninum (sample Nc-1), for the use in immunodiagnostic tests as immunofluorescence, immunohistochemistry, ELISA and dot-ELISA. For the production of mAb the sonicated protozoan was used, originating from the culture in cells VERO and it was purified by filtration, it was used the tachyzoites for the immunization of the mice BALB/c, using as saponin adjuvant , what allowed the obtaining of polyclonal antibodies. The fusing of the esplenic cells of the immunized mice and the myelomas SP2/0 resulted in the obtaining of 72,4% of hybridomas secretors of antibodies anti-Nc-1. After the cloning for limiting dilution was obtained 78,2% of the clones secretors of antibodies anti-Nc- 1. After the recloning for limiting dilution, they were obtained 4 clones secretors of mAb of the class IgG. Obtained mAb IgG2a were capable to reveal the proteins of mass molecular 25 kDa and 70 kDa for "western blotting" to inhibit the multiplication of the taquyzoites in cells VERO. The monoclonal antibodies produced can be used in the development of more sensitive tests, in the epidemic studies of the neosporosis or they can be used in the identification of relevant proteins for the development of vaccines. |
publishDate |
2009 |
dc.date.available.fl_str_mv |
2009-08-03 2015-03-26T13:46:40Z |
dc.date.issued.fl_str_mv |
2009-02-20 |
dc.date.accessioned.fl_str_mv |
2015-03-26T13:46:40Z |
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.citation.fl_str_mv |
DEVENS, Bruna Alves. Production of hybridomas and clones to obtain monoclonal antibodies anti - Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae). 2009. 3 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2009. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/4977 |
identifier_str_mv |
DEVENS, Bruna Alves. Production of hybridomas and clones to obtain monoclonal antibodies anti - Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae). 2009. 3 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2009. |
url |
http://locus.ufv.br/handle/123456789/4977 |
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Universidade Federal de Viçosa |
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Mestrado em Medicina Veterinária |
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UFV |
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BR |
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Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de |
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Universidade Federal de Viçosa |
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