Production and Characterization of IgY Polyclonal Antibodies Specific to Human Interleukin-6 and Their Neutralization Potential for Anti-inflammatory Responses

Abstract

Background: Interleukin-6 plays an essential role in cytokine storm and cytokine release syndrome, which occur in response to pathogen infection or tissue injury and are associated with severe symptoms. Neutralizing IL-6 can help reduce symptom severity. Chicken eggs serve as an excellent alternative antibody source compared to mammalian serum. The immunoglobulin Y (IgY) in the chicken’s blood is transferred to and depos-ited within the egg yolk in large amounts. Several IgY products have been developed for therapeutic applications in various diseases. This study focuses on producing anti-human IL-6 (IL-6) IgY antibodies to support therapeutic advancements. Methods: Anti-IL-6 IgY was generated by immunizing three hens with recombinant IL-6 protein mixed with alum adjuvant, immunized four times at three-week intervals. Then, IgY was extracted from egg yolks. The specificity of IgY was determined by Western blot. The neutralizing activity against secreted IL-6 was demonstrated by Human Coronavirus OC43 (HCoV-OC43)-infected cells and Lipopolysaccharide (LPS)-stimulated human lung fibroblast MRC-5 cells. Results: The specific anti-IL-6 was detected starting from day 10 to day 90 after im-munization. The average yield of total IgY was 17.97±15.66 mg per egg. The extracted anti-IL6 IgY antibody exhibited efficient neutralizing effects against secreted IL-6 in the HCoV-OC43-infected or LPS-stimulated cells in a dose-dependent manner. Conclusion: This study highlights the ease of production and the satisfactory yield of anti-IL-6 IgY derived from chicken eggs. The antibody demonstrates an in vitro inhibi-tory effect on IL-6, with potential applications in therapeutic development.