The aims of this research are confirming the feasibility of hybrid immunoglobulin-binding reagents, its used in ELISAs for IgG/IgY detection and detecting specific antibodies against an infectious microorganism (Salmonella spp) in various animal species using a universal diagnostic ELISA. Hybrid immunoglobulin-binding bacterial proteins (IBP) including recombinant protein LA, recombinant protein LG, and recombinant protein AG have been produced for improvement of their binding affinity to a much larger number of immunoglobulins. This hybrid bacterial protein thus represents a powerful tool for binding, detection, and purification of immunoglobulins and their fragments. However, SpLA-LG-peroxidase and SpLAG-anti-IgY-peroxidase were produced by the periodate method. They have shown to be effective reagents. Their binding affinity to immunoglobulins surpasses previous hybrid IgG-binding proteins reported, including the most known SpAG, SpLA and SpLG. The IgY fraction was isolated from the egg yolks of a variety of birds including species of chicken, bantam hen, guinea hen, quail, goose, duck, wild and domestic pigeon, parakeet, cattle egret, pheasant, and ostrich. The IgY fraction was isolated by the chloroform-polyethylene glycol (PEG) method. An ELISA for anti-Salmonella spp antibodies was employed with some modifications to determine the presence of antibodies in humans, laying hens, geese, quails, and pigeons. Salmonella are motile, flagellated rod-shaped zoonotic pathogens which may survive with or without oxygen. They belong to the family Enterobacteriaceae and is implicated with typhoid fever and food-borne illnesses. This pathogen is associated with several diseases, which may become fatal and negatively impact the health of individuals and various economies globally. The poultry industry is most impacted and vulnerable to the onslaught of this pernicious microbe.

Background: IgM, which participates in the primary immune response, is the primary antibody in egg whites. There is scant information about the production of antibodies in egg whites. This study describes the preparation of antibodies against bacterial antigens. Methods: Enzyme-linked immunosorbent assay (ELISA) was used to detect the presence of anti-egg white antibodies. The antibodies were purified using affinity chromatography. Statistical analyses were performed using SPSS version 22. Statistical significance was set at P<0.05. Results: Large amounts of anti-protein A antibodies were produced in chicken egg whites. The generation of anti-SpA antibodies was demonstrated by affinity chromatography from 9 d post-immunization egg white samples. Inhibition of agglutination was observed in samples containing anti-SpA antibodies, and agglutination at the bottom of the wells was observed in the negative samples. Conclusion: The use of hyper-immune egg whites as an option for antibiotic use could be advantageous for treating infectious diseases because of the large number of antibodies produced, reducing antigenic variation, low cost, and toxicity.

Isolation of antibodies from the egg yolk of chickens is of particular interest as a source of specific antibodies for oral administration to prevent infections and use them as immunodiagnostic reagents. The use of birds in antibody production results in a reduction in the use of laboratory animals. Immunized chickens produce larger quantities of antibodies (2000 mg IgY/month) than rodents (200 mg IgG/month) in the laboratory. According to Jerne’s network theory, it is possible to produce an antibody against the antigen-binding site of another antibody. This study assessed the hypothesis that immunization with viral peptides (immunogens) could provide a potent immune response that could be evaluated in chicken eggs. Human immunodeficiency virus 1(HIV-1) is used as an immunogen. The second hypothesis was that an orally administered antibody stimulates the production of a complementary antibody, the so-called anti-idiotypic antibody, which can potentially be therapeutical. This study reports and analyzes the use of eggs as therapeutic agents. We wanted to test the hypothesis that feeding chicks with hyperimmune eggs stimulates the production of anti-anti-idiotypic antibodies that neutralize the original HIV antigen fragments 308-331 or 421-438 of gp120 or fragment 579-601 of gp41. Future research could entail an anti-idiotype strategy for prophylactic vaccines. It is vital to note that it may need an anti-idiotype response to prime immunity against an HIV viral epitope, which may be used as a secondary element. The use of anti-idiotype immune responses in infected individuals may shift the balance of the immune system, allowing the organism to manage HIV infection. Therefore, it may be an avenue for immunotherapy to improve the fight against HIV infections. However, more studies and clinical trials are required to demonstrate similar human immune responses as observed in birds.

IFN-γ is a key player in driving cellular immunity. It is capable of orchestrating numerous and vital protective functions to enhance immune responses in cancers and infectious diseases. It exhibits its immunomodulatory effects by enhancing antigen processing and presentation,, inducing an anti-viral state, increasing leukocyte trafficking, boosting the anti-microbial properties of immune cells and by affecting cellular proliferation and apoptosis. [1] Reference

Angel A Justiz-Vaillant, Belkis Ferrer-Cosme 2020. ELISA for quantification of CXC motif chemokine ligand 2 (CXCL2) in human serum or plasma.. protocol s.i o protocol s.i o