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Permanent link (DOI): https://doi.org/10.7939/R3V693

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Antibody mediated “Universal” Osteoclast targeting platform Open Access

Descriptions

Other title
Subject/Keyword
Monoclonal antibody
Osteoclast
RANK receptor
Single chain Fraction variable
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Newa,Madhuri
Supervisor and department
Doschak,Michael ( Faculty of Pharmacy and Pharmaceutical Sciences)
Examining committee member and department
Jurasz, Paul (Faculty of Pharmacy and Pharmaceutical Sciences)
Löbenberg,Raimar (Faculty of Pharmacy and Pharmaceutical Sciences)
Wishart, David (Department of Biological Sciences)
Krawetz,Roman ( Faculty of Medicine)
Kaur,Kamaljit (Faculty of Pharmacy and Pharmaceutical Sciences)
Doschak,Michael ( Faculty of Pharmacy and Pharmaceutical Sciences)
Department
Faculty of Pharmacy and Pharmaceutical Sciences
Specialization
Pharmaceutical Sciences
Date accepted
2013-07-10T10:03:52Z
Graduation date
2013-11
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Osteoclasts are the sole cells responsible for bone resorption and their activity is central to the process of bone remodeling. Excessive osteoclast activity leads to increased bone resorption, predisposing individuals to bone conditions such as osteoporosis, Paget’s disease and the focal bone erosions seen in rheumatoid arthritis. Hence, the pharmacological arrest of osteoclasts is the mainstay of treating many bone diseases. However, none of the current antiresorptive therapies for bone loss target osteoclast cells directly. As osteoclasts express the Receptor Activator of Nuclear factor Kappa B (RANK) receptor, the essential signaling receptor for osteoclast differentiation, it was hypothesized that antibody-like molecules generated with specificity against RANK could selectively target and deliver conjugated drug cargo to the osteoclast. The objectives of this thesis were to generate, characterize and evaluate monoclonal antibodies against the osteoclast RANK receptor, in order to develop a pharmaceutical platform capable of selective and targeted drug delivery to osteoclast cells. Using hybridoma technology, a specific monoclonal antibody against recombinant human RANK receptor was generated. Synthesis of osteoclast-targeting bioconjugates with that antibody was conducted using the antiresorptive peptide hormone calcitonin. Conjugate characterization was undertaken and its efficacy tested on osteoclast cell cultures using various osteoclast specific assays. Both the conjugate as well as the antibody itself showed remarkable inhibition of osteoclast activity. The desirable result obtained with the mere binding of the antibody to the receptor led us towards new research work, focusing on valuable scale-up production and therapeutic use. As a new research direction, single-chain Fraction variable (scFv) antibody-like molecules was expressed against the human RANK receptor using phage display technology, to circumvent complications associated with murine-derived antibodies. The Anti-RANK scFv showed specificity towards osteoclast RANK receptors and also showed an inhibitory effect on osteoclast activity. With the increase in development trends for biologics as therapeutics and the growing knowledge on the importance of osteoclast targeted therapy, this novel biologic reagent may find utility as “universal osteoclast targeting platform”. This may provide a meaningful strategy in terms of osteoclast targeting and drug delivery with the aim of treating or controlling the progression of osteoclast related bone disorders.
Language
English
DOI
doi:10.7939/R3V693
Rights
Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
Citation for previous publication
Newa, M.; Bhandari, K. H.; Tang, L.; Kalvapalle, R.; Suresh, M.; Doschak, M. R.: Antibody-mediated "universal" osteoclast targeting platform using calcitonin as a model drug. Pharm Res 2011, 28, 1131-43.Bhandari, K.H., Newa, M., Chapman, J. & Doschak, M.R. (2012) Synthesis, characterization and evaluation of bone targeting salmon calcitonin analogs in normal and osteoporotic rats. J Control Release, 158, 44-52

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