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Ultrasensitive Point-of-Care Dengue Diagnostics and Vaccine Applications Open Access

Descriptions

Other title
Subject/Keyword
Dengue
Bispecific Antibody
Immunoswab
Monoclonal Antibody
Chicken IgY
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Ganguly, Advaita
Supervisor and department
Loebenberg, Raimar (Faculty of Pharmacy and Pharmaceutical Sciences)
Sunwoo, Hoon H (Faculty of Pharmacy and Pharmaceutical Sciences)
Examining committee member and department
Siraki, Arno (Faculty of Pharmacy and Pharmaceutical Sciences)
Doschak, Michael (Faculty of Pharmacy and Pharmaceutical Sciences)
Careem, Faizal A (Veterinary Medicine, Universit of Calgary)
Department
Faculty of Pharmacy and Pharmaceutical Sciences
Specialization
Pharmaceutical Sciences
Date accepted
2013-09-29T20:03:40Z
Graduation date
2013-11
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Dengue virus infections can result in a range of clinical manifestations from asymptomatic infection to dengue fever and the severe disease dengue haemorrhagic fever/dengue shock syndrome. The disease is now endemic in more than 100 countries in Africa, the Americas, the eastern Mediterranean, Southeast Asia, and the Western Pacific, threatening more than 2.5 billion people. The World Health Organization estimates that there may be 50 million to 100 million cases of dengue virus infections worldwide every year, which result in 250,000 to 500,000 cases of Dengue Hemorrhagic fever and 24,000 deaths each year. The dengue virus non-structural NS1 protein is a 46–50 kDa glycoprotein when expressed in infected mammalian cells. A high circulating level of NS1 was demonstrated in the acute phase of dengue infection by antigen capture ELISAs. The precise function of dengue NS1 protein remains unclear. However, antigen detection of non-structural dengue antigens may be of benefit for an early stage rapid diagnosis of infection due to its long half-life in the blood. Five high affinity monoclonal antibodies were developed and characterized against the recombinant dengue NS1 protein using hybridoma technology. Anti-NS1 and anti-HRPO hybridomas were fused and sorted to develop a series of bi-specific antibodies. The recombinant NS1 protein was also used to immunize chickens for the development of Anti-NS1 chicken IgY polyclonal antibody. The different combinations of anti-NS1 mAbs, bi-specific antibodies and chicken IgY were used in the development of simple, rapid, inexpensive, highly sensitive, specific and easy to perform assays for the detection of dengue virus infection. The dengue envelope protein was also expressed using recombinant techniques for the evaluation of its role as a vaccine candidate. The envelope protein is known to induce neutralizing antibodies against dengue virus. The recombinant dengue envelope protein was used to immunize mice as a vaccine candidate. Serum antibody analysis confirmed virus neutralization characteristics. We also raised monoclonal antibodies against the envelope protein to better understand dengue pathogenesis as well as potential reagents in dengue diagnostic development.
Language
English
DOI
doi:10.7939/R38G8FV0S
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.
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