versión On-line ISSN 1025-0298
REYES, Gisele et al. Detection of fibronectin and collagen binding-proteins from Leptospira interrogans serovar Canicola. Vaccimonitor [online]. 2007, vol.16, n.3, pp. 1-6. ISSN 1025-0298.
ABSTRACT As part of the studies conducted to obtain a vaccine formulation by subunits against human leptospirosis, purification and characterization of fibronectin and collagen binding-proteins in Leptospira interrogans was developed. Outer membrane proteins from Leptospira were solubilized with Triton X-114 and were applied in a first step into an IgG AntiBSA- Sepharose 2B CL affinity chromatographic column to eliminate BSA as main contaminant of culture media for this microorganism. The BSA free fraction was applied into a Fibronectin-Sepharose 4B CNBr affinity column. A fibronectin binding protein from Leptospira interrogans serovar Canicola strain 87, with a molecular weight of 40 kDa was obtained. For this, specific sera obtained from experimentally infected rats with each serovar and mixes of human sera of convalescent patients were used. This protein showed to be antigenic and conserved in Canicola, Copenhageni and Mozdok serovars. Solubilized outer membrane protein by Triton X114 was applied into another affinity column of Collagen-Sepharose 4B CNBr to isolate collagen binding proteins from Leptospira interrogans serovar Canicola strain 87. A collagen binding protein with a molecular weight of 25 kDa which showed to be antigenic when evaluated by Dot-Blot against human sera of convalescent patients was found.
Palabras llave : Leptospira; collagen; fibronectin; affinity chromatography.