Preparation of Anti-RANKL IgY for Chicken Bone Health

• Author / Creator

  Sharma, Kumakshi

• Osteoporosis is one of the prevalent disorders that exist in both humans and chickens. The bone disorder is characterized by depletion of bone mineral content, leading to fragile bones and increases susceptibility to fractures. During the egg laying cycle, cage layers are more prone to fragile bones and fractures, due to lack of physical activity, nutritional factors and bone resorption. Receptor activator of nuclear factor kappa B Ligand (RANK-L) expressed on the surface of osteoblasts play a major role in bone resorption process. Interaction and binding of RANK-L to its receptor (RANK) on the surface of osteoclasts, results in differentiation and maturation of osteoclasts, hence, leading to osteclastogenesis. This thesis focuses on development of therapeutic that would inhibit the activity of RANKL and prevent bone breakdown in chickens. Immunoglobulins in chicken egg yolk, calledIgY have been reported to be potential candidate against several bacterial pathogens. Producing IgY against RANK-L would be an effective way to treat osteoporosis in chickens by binding to RANK-L, hence preventing bone resorption caused by RANK – RANK-L interaction. The aim of this project is to produce recombinant chicken RANK-L using E. Coli and Baculovirus expression systems. Because the expression of chicken RANK-L, E. Coli expression system led to formation of insoluble inclusion bodies, there were challenges to solubilize and purify the proteins. However, baculovirus expression system was successfully able to produce intracellular recombinant ligands. The chickens were immunized intramuscularly with chicken RANK-L, and then IgY specific to RANK-L was isolated and purified from egg yolk. The affinity and binding of specific IgY with respective RANK-L was investigated and showed by immunoasays. A preliminary in vitro study was done on murine marcrophage cells (RAW 246.7). The cells showed positive tartrate-resistant acid phosphatase activity when the marcrophages were treated with mouse and chicken RANK-L and there was no activity when cells were treated with RANK-L and their specific IgY.

• Subjects / Keywords

  • Osteoporosis, Chickens

• Graduation date

  Fall 2018

• Type of Item

  Thesis

• Degree

  Master of Science

• DOI

  https://doi.org/10.7939/R3JH3DJ46

• License

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