SciELO - Scientific Electronic Library Online

 
vol.27 número2Cholinesterases in total blood measured with a semiquantitative technique, and plasma or erythrocyte cholinesterases measured with quantitative techniques índice de autoresíndice de assuntospesquisa de artigos
Home Pagelista alfabética de periódicos  

Serviços Personalizados

Journal

Artigo

Indicadores

Links relacionados

  • Em processo de indexaçãoCitado por Google
  • Não possue artigos similaresSimilares em SciELO
  • Em processo de indexaçãoSimilares em Google

Compartilhar


Biomédica

versão impressa ISSN 0120-4157versão On-line ISSN 2590-7379

Resumo

CASTANEDA, Nadia Yadira; CHAPARRO-OLAYA, Jacqueline  e  CASTELLANOS, Jaime E. Production and characterization of a polyclonal antibody against rabies virus phosphoprotein. Biomédica [online]. 2007, vol.27, n.2, pp.257-267. ISSN 0120-4157.

Introduction. The expression of recombinant viral proteins has been a useful tool to study molecular biology and pathogenesis of virus infections. Because commercial specific antibodies to rabies virus phosphoprotein (P) are currently unavailable, these antibodies must be generated de novo in order to study the role of P protein during the infectious process. Objective. A polyclonal antibody was produced and characterized for use against the phosphoprotein (P) of rabies virus. The antibody was raised in rabbits with a recombinant viral phosphoprotein (P) produced in Escherichia coli. Materials and methods. Gene P coding for the viral phosphoprotein (P) was amplified by RT-PCR and cloned into the expression vector PinPointTM Xa-1 T. The recombinant protein P was expressed in Escherichia coli, purified by affinity chromatography and used to produce a polyclonal antibody anti-P. The antibody anti-P was purified and characterized by immunocytochemistry, immunofluorescence, fluorometric CELL-ELISA and Western blotting. Results. The recombinant viral phosphoprotein was successfully expressed as a 50 kd biotinylated fusion protein which corresponds to the whole protein P of rabies virus. The polyclonal antibody raised against this recombinant protein P was able to detect with high specificity, protein P in cultures of sensorial neurons infected with rabies virus. Conclusions. The P protein obtained from heterologous expression in Escherichia coli became a specific antigen that was used to produce a polyclonal antibody capable of detecting native P protein in rabies virus infected cells.

Palavras-chave : rabies virus; immunochemistry; phosphoproteins; recombinant proteins; Escherichia coli.

        · resumo em Espanhol     · texto em Espanhol     · Espanhol ( pdf )

 

Creative Commons License Todo o conteúdo deste periódico, exceto onde está identificado, está licenciado sob uma Licença Creative Commons