INGEBI   02650
INSTITUTO DE INVESTIGACIONES EN INGENIERIA GENETICA Y BIOLOGIA MOLECULAR "DR. HECTOR N TORRES"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
POSITIONAL SCANNING SYNTHETIC LIBRARIES TO THE STUDY OF T. cruzi RECOMBINANT HUMAN ANTIBODY SPECIFICITY
Autor/es:
L NIBORSKI; V JUDKOWSKI V ; C PINILLA; A MENTABERRY; KA GOMEZ; MJ LEVIN
Lugar:
Puerto Madryn
Reunión:
Congreso; XLVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular; 2010
Institución organizadora:
SAIB
Resumen:
It is well established that protective immune mechanisms against T. cruzi are constituted in part by strong antibody (Ab) responses to the parasite. However, it has also been proposed that some parasite Abs can cross-react to human antigens, suggesting that autoimmunity may have a role in the pathogenesis of Chagas disease. The aim of this study was to elucidate the antigen specificity of relevant human T. cruzi antibodies. Using phage display technology, two single chain variable fragments (scFvs) 6B and A1 were isolated from Chagas heart Disease patients. Western-blot analysis revealed that scFv 6B and scFv A1 recognized T. cruzi proteins of 175 and 47 kDa respectively. Interestingly, scFv A1 also identified a 47 kDa protein in mouse brain lysate. The antigen specificity of scFv 6B and scFv A1 was examined using a positional scanning hexapeptide library, which was screened by competitive ELISA for its ability to inhibit Ab binding to a parasite lysate. This proteomic information together with a positional scanning based biometrical analysis that systematically compares the screening results of the library with selected protein databases will result on the identification of candidate T. cruzi antigens as well as cross-reactive antigens from the human and murine species.