Osteoporosis is a skeletal disorder that affects millions of people worldwide, and is characterized by the accelerated loss of bone mass. Current anti-resorptive drug approaches such as bisphosphonates and salmon calcitonin exhibit severe side effects and very low bioavailability, respectively. In this study, we have designed, synthesized, and performed preliminary tests on a novel conjugate that targets the RANK receptor on bone resorbing cells (osteoclasts) in vitro with one arm, while delivering a specific effector molecule, calcitonin, to osteoclasts with the other arm. First, we successfully generated osteoclasts from precursor RAW 264.7 cells and confirmed that they were functional. We also designed a resorption assay that can be used to test the efficacy of new and existing anti-resorptive drugs. RAW 264.7 cells were then treated with an antibody to RANK to prove that anti-RANK could be used as a targeting mechanism. We then showed that delivery of calcitonin-loaded anti-Calcitonin antibodies onto osteoclasts results in the association of calcitonin onto its receptors on osteoclasts. Finally, we constructed a novel conjugate: calcitonin–Streptavidin–anti-RANK, and showed that it can be used to introduce calcitonin into an osteoclast-like microenvironment.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:AEU.10048/443 |
| Date | 11 1900 |
| Creators | Kalvapalle, Rohit |
| Contributors | Doschak, Michael (Pharmacy and Pharmaceutical Sciences), Suresh, Mavanur (Pharmacy and Pharmaceutical Sciences), Kane, Kevin (Medical Microbiology and Immunology), Matyas, John (Cell Biology and Anatomy, University of Calgary) |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | en_US |
| Detected Language | English |
| Type | Thesis |
| Format | 1742876 bytes, application/pdf |
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