Produção de hibridomas e clones produtores de anticorpos monoclonais anti- Nucleosídeo trifosfato difosfohidrolase 1 (NTPDase-1) recombinante de Trypanosoma cruzi

• Ano de defesa: 2010
• Autor(a) principal: Valle, Myrian Augusta Araújo Neves do
• Orientador(a): Fietto, Juliana Lopes Rangel
• Banca de defesa: Silva Júnior, Abelardo , Fietto, Luciano Gomes , Siqueira-batista, Rodrigo , Sossai, Sidimar
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• Idioma: por
• Instituição de defesa: Universidade Federal de Viçosa
• Programa de Pós-Graduação: Mestrado em Biologia Celular e Estrutural
• Departamento: Análises quantitativas e moleculares do Genoma; Biologia das células e dos tecidos
• País: BR
• Palavras-chave em Português: NTPDase-1; Hibridoma; Anticorpos monoclonais; T. cruzi
• Palavras-chave em Inglês: NTPDase-1; Hybridoma; Monoclonal; T. cruzi
• Área do conhecimento CNPq: CNPQ::CIENCIAS BIOLOGICAS::BIOLOGIA GERAL
• Link de acesso: http://locus.ufv.br/handle/123456789/2330

The protozoan Trypanosoma cruzi is the etiologic agent of Chagas disease. The ecto-nucleotidases E-NTPDase family are enzymes that hydrolyze nucleotide tri-and / or diphosphate monophosphates in their products. The NTPDase-1 T. cruzi is a protein present on its surface, with the possibility of also being secreted into the extracellular milieu. Its function is still not completely understood, but recent data strongly suggest its involvement in virulence and infectivity of the parasite is considered a good target for blocking infection. Due to the difficulty of diagnosis in chronic phase, morbidity and disability caused by disease treatment, it is extremely important to developing new and more efficient methods of diagnosis and treatment, in which case the monoclonal antibodies a powerful tool. The purpose of this work was the production of monoclonal antibodies that specifically recognize the NTPDase-1 T. detriment cruzi NTPDase-1 Leishimania infantum chagasi. For assessing the expression of heterologous NTPDase-1 in bacterial system (E. coli BL-21) and its purification by affinity chromatography on nickel-agarose column (Ni-NTA). It was used for the immunization of mice to protein obtained, with the confirmation and titration of hyperimmune serum made by dot-ELISA. B lymphocytes from immunized mice were fused with myeloma cells (SP2 / 0) to obteção hybridoma, and 47.13% of hybridomas secreting anti-NTPDase-1 T. cruzi, the hybridoma were selected by ELISA methodo. After limiting dilution, 49 clones recognized the NTPDase-1 recombinant T. cruzi which represents approximately 27% of positive clones. In addition 10 clones also recognized the NTPDase-1 Leishmania infantum chagasi. Therefore, we isolated 39 clones that specifically recognized only the NTPDase-1 recombinant T. cruzi.