Produção de hibridomas e clones para obtenção de anticorpos monoclonais anti-Neospora caninum Nc-1 (Apicomplexa, Sarcocystidae)

• Ano de defesa: 2009
• Autor(a) principal: Devens, Bruna Alves
• Orientador(a): Viloria, Marlene Isabel Vargas
• Banca de defesa: Guimarães, Antonio Marcos , Sales Junior, Policarpo Ademar , Oliveira, Leandro Licursi de
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Viçosa
• Programa de Pós-Graduação: Mestrado em Medicina Veterinária
• Departamento: Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de
• País: BR
• Palavras-chave em Português: Neospora caninum; Anticorpos monoclonais
• Palavras-chave em Inglês: Neospora caninum; Monoclonal antibodies
• Área do conhecimento CNPq: CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::PATOLOGIA ANIMAL
• Link de acesso: http://locus.ufv.br/handle/123456789/4977

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.