Resumo

PADILLA-LONDONO, Jonathan Mauricio; TREJOS-SUAREZ, Juanita; CORONADO, Sandra Milena  e  CARDONA-PEREZ, Néstor Iván. Production of Polyclonal Antibodies against Toxoplasma gondii Recombinant protein with Nitric Oxide Synthase activity and immunologic marking in Tachyzoites. Infect. [online]. 2008, vol.12, n.2, pp.111-119. ISSN 0123-9392.

The nitric oxide synthase (NOS)is an enzyme well described on mammals but little is known about the role of these enzymes on pathogenic parasites. We produced polyclonal antibodies against a recombinant NOS enzyme from Toxoplasma gondii nd e lso er formed n mmunol locali zation of the enzyme on tachyzoites. We used two New Zealand rabbits (Oryctolagus cuniculus) to perform intramuscular immunization and we used two types of adjuvant: aluminum hydroxide and Freund ’s adjuvant. We tested the generation of polyclonal antibodies by an indirect ELISA assay. The results showed that levels of antibodies were higher in rabbits immunized with Freund ’s adjuvant than with aluminum hydroxide (p =0.005). In order to test cross reactions, we used a recombinant Toxoplasma metallooprotease, a recombinant cysteine protease from Entamoeba histolytica, albumin 2%, hydroxide aluminium and Freund ’s adjuvant as antigens on indirect ELISA assays for the polyclonal serum antibodies. No serum showed absorbances higher than the cutoff level with these antigens, indicating that the polyclonal antibodies were specific for recombinant Toxoplasma NOS. Additionally, we performed immunodetection of the enzyme on Toxoplasma gondii tachyzoites and we obtained a diffuse labeling of the parasite.

Palavras-chave : Toxoplasma gondii; nitric oxide synthase; policlonal antibodies; aluminum hydroxide; Freund´s adjuvant.

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