Antibodies anti-Shiga toxin 2 B subunit from chicken egg yolk: isolation, purification and neutralization efficacy

• Autores: Parma, Y. R.; Chacana, Pablo; Rogé, Ariel; Kahl, A; Cangelosi, Adriana; Geoghegan, Patricia A.; Lucchesi, P. M. A.; Fernández-Miyakawa, Mariano E
• Año de publicación: 2011
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: Toxinas Shiga; Antitoxinas; Pollos; Neutralización
• Institución: Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"

Fil: Parma, Y. R. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Instituto de Patobiología; Argentina.
Fil: Chacana, Pablo. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Instituto de Virología; Argentina.
Fil: Rogé, Ariel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Productos de Biológicos. Servicio Sueros y Antígenos; Argentina.
Fil: Kahl, A. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Instituto de Virología; Argentina.
Fil: Cangelosi, Adriana. ANLIS Dr.C.G.Malbrán. Centro Nacional de Control de Calidad de Biológicos. Servicio de Inmunoterapéuticos; Argentina.
Fil: Geoghegan, Patricia A. ANLIS Dr.C.G.Malbrán. Centro Nacional de Control de Calidad de Biológicos. Servicio de Inmunoterapéuticos; Argentina.
Fil: Lucchesi, P. M. A. Universidad Nacional del Centro. Facultad de Ciencias Veterinarias. Departamento de Sanidad Animal y Medicina Preventiva. Laboratorio Inmunoquímica y biotecnología; Argentina.
Fil: Fernández-Miyakawa, Mariano. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias. Instituto de Patobiología; Argentina.
Shiga toxins (Stx1 and Stx2) are the main virulence factors of enterohemorrhagic Escherichia coli (EHEC), a foodborne pathogen associated with diarrhea, hemorrhagic colitis and hemolytic uremic syndrome. The aim of this study was to evaluate the antibodies against Stx2 obtained from egg yolks of laying hens immunized with a recombinant Stx2B subunit. A high specific response in serum was observed 25 days after the first immunization and IgY antibodies were extracted from day 47th and purified from egg yolk. A concentration of 0.84 mg of total IgY/ml of egg yolk was obtained, of which 8% were antigen specific. The ability of anti-Stx2B IgY to recognize Stx2B and Stx2 either in solid-phase or in solution were evaluated and compared with anti-Stx2B rabbit antibodies by Western blotting and ELISA. The protective efficacy of IgY against Stx2 was determined by in vitro and in vivo experiments. The results show that IgY was able to recognize Stx2B and Stx2 in denatured conditions, attached to a solid-phase and free in solution. The anti-Stx2B IgY could effectively block the biological activity of Stx2 on Vero cells and protect mice from Stx2 challenge. The data suggest that immunization of hens with Stx2B could be a strategy to obtain at low cost a relatively high concentration of anti-Stx2 egg yolk IgY, able to neutralize Stx2 lethal activity. IgY technology could be an useful tool for research, diagnosis and therapy of EHEC infection.