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Permanent link (DOI): https://doi.org/10.7939/R33F4KW7V

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Rational Design of an HIV-1 Vaccine using a Filamentous Phage Carrier Open Access

Descriptions

Other title
Subject/Keyword
filamentous phage
HIV-1
glycoconjugate vaccine
Type of item
Thesis
Degree grantor
University of Alberta
Author or creator
Bailey, Justin, J.
Supervisor and department
Bundle, David R.
Examining committee member and department
Cairo, Chris (Chemistry)
Klassen, John S. (Chemistry)
Meyer, Bernd (Chemistry)
Campbell, Robert E. (Chemistry)
Department
Department of Chemistry
Specialization

Date accepted
2013-04-02T14:04:05Z
Graduation date
2013-06
Degree
Doctor of Philosophy
Degree level
Doctoral
Abstract
Glycans are a diverse and integral component of all biological organisms, making them an ideal target for vaccine development. Carbohydrates often act as weak antigens, lacking the T cell epitopes required to recruit T cell help which is vital to develop a strong and long lasting protective antibody response. Chemical conjugation of glycan antigens to a protein carrier to elicit T cell help has been a successful strategy to improve the strength of the immune response towards glycan-derived haptens. The hapten-carrier concept has been successfully used for numerous synthetic glycoconjugate vaccines now approved for human use. HIV-1 appears to be an ideal candidate for developing a glycoconjugate vaccine as the exposed HIV-1 envelope protein, gp120, is one of the most heavily N-glycosylated proteins known. Glycoconjugate vaccines displaying the glycans of gp120 have thus far failed to elicit protective antibodies that bind to gp120 or neutralize viruses bearing HIV 1 Env. The lack of cross reactivity has been attributed to an inability of the glycoconjugate vaccines to faithfully mimic the carbohydrate-dense gp120 surface. Described herein is the synthesis of two mannosyl thiol analogues of gp120 glycans, their chemical conjugation to a filamentous phage carrier, and the LC-UV-MS methodology developed to characterize the resulting glycoconjugates. The mannosyl thiols provide access to highly reactive linker chemistry while maintaining a minimal linker length, which may be essential to mimic the limited flexibility of the tightly packed glycans of gp120. Filamentous phage possess a structurally homogenous, repeating surface with closely spaced, solvent exposed amines available for conjugation. This arrangement is ideal for mimicking the glycan shield of gp120. Coupling of mannosyl thiol analogues to the phage carrier produced a densely glycosylated surface where over 80% of the 8,100+ conjugation sites of the phage were occupied, as was determined to an unprecedented level of clarity by LC-UV-MS. The phage glycoconjugates were shown to interact with anti-HIV-1 glycan-specific antibody 2G12 with 0.5 nM affinity, ranking it as one of the best 2G12-binding antigenic mimics described thus far. Mice immunized with the phage glycoconjugates elicited high titres of carbohydrate-specific antibodies that were unable to cross react with gp120.
Language
English
DOI
doi:10.7939/R33F4KW7V
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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