Generation and Characterization of IgY antibodies from Lohmann Brown Hens Immunized with Salmonella spp. for their Subsequent Application in Nanotherapy

Abstract

Salmonellosis is a common type of food poisoning caused by the Salmonella enterica bacterium. This disease is common in Ecuador and it is propagated through poultry. A new alternative for controlling the infection in humans and animals is through passive immunity. Consequently, specific antibodies can be developed as therapeutic agents based on biodegradable nanoparticles liposomes. Specific antibodies are produced by inoculation of hens with specific antigens, followed by retrieving the antibodies accumulated in yolk. The objective of this work was to purify specific IgY antibodies against Salmonella spp from Lohmann Brown hens. A group of hens was immunized with a pool of Salmonella enterica subsp enterica serovar Enteritidis and Salmonella enterica subsp enterica serovar Infantis. These two pools of bacteria were previously inactivated with a treatment (T1) by forlmaldehyde or with a treatment (T2) by heat. Lipids were removed using 0.1% pectin, the protein pelleted with ammonium sulfate (35% w/v) is re-suspended in PB buffer 0.025M pH 8. The Total IgY antibodies were purified by DEAE cellulose. The highest yield of total IgY was 5.5 mg IgY/mL yolk following the (T2) treatment at 63 days post-immunization. The characterization was performed with ELISA and MABA techniques and the immunoreactivity was evaluated by Western blot in total antisera and purified protein, hybridized with commercial anti-chicken antibodies from SIGMA. The heavy chain 67 kDa and the light chain 25 kDa were visualized. Such antibodies could able to ensure quick and immediate protection against several biotargets, the new IgY has potential for nanotherapy against Salmonelosis diseases.