EGG YOLK IMMUNOGLOBULIN AGAINST PSEUDOMONAS AERUGINOSA OPRF, PREVENTION AND TREATMENT OF INFECTIONS

Background and Aim:Nosocomial infection is a main problem for patients after hospitalization. Pseudomonas aeruginosa is an opportunistic bacteria which can cause these hospital acquired infections. To prevent development of novel resistant strains, antibiotic therapy should be replaced with other methods. Based on existing condition, antibody utilization can be an appropriate approach. In comparison to other antibodies, egg yolk immunoglobulin (IgY) is a reasonable choice as it has many advantages like easy production and extraction procedure.Methods:In this study hens were immunized intramuscularly with OprF recombinant protein which is highly conserved among various P.aeruginosa strains. IgY was extracted from egg yolks in two stages. First, egg yolks diluted 7 times (by volume) with water, pH adjusted to 5.0 with 0.5 M HCl, and the mixture was frozen at -20 °C. The frozen mixture was transferred to a funnel with conventional filter paper and allowed to melt freely at room temperature. In the second isolation stage, IgY precipitated with 8.8% w/v NaCl, pH was adjusted with 0.5 M HCl to 4.0. Mixtures were stirred for 2 hours at room temperature and then centrifuged at 3,380 g for 20 min at 4 °C. Supernatants were discarded and the pellets dissolved in PBS. The antibody titers were measured by ELISA technique.Results:ELISA results showed that antibody titers significantly increased in comparison to control samples and the produced immunoglobulin had the ability of detection and attachment to the OprF.Conclusion:The obtained immunoglobulin could be an effective choice for passive immunotherapy against P.aeruginosa infections.

Keywords:

Pseudomonas aeruginosa , OprF , IgY