Graduation Date

Fall 12-16-2022

Document Type

Dissertation

Degree Name

Doctor of Philosophy (PhD)

Programs

Pharmaceutical Sciences

First Advisor

Yuri L. Lyubchenko

Second Advisor

Joseph A. Vetro

Third Advisor

Yuxiang Dong

Fourth Advisor

Luis A. Marky

MeSH Headings

Vaccine, nanotechnology, Alzheimer's, coronavirus, epitope, modular

Abstract

Vaccines are among the most effective and safe therapeutics known to mankind. Historically the approach to the creation of new vaccines has been dominated by empirical research, trial, and error. This thesis focuses on the more modern, rational approaches to vaccinology based on the current understanding of the inner workings of the immune system. We used nanotechnology for the generation of novel immunogens, addressed several challenges, and characterized the immune responses.

The preparation, characterization, and troubleshooting of several vaccines against dimeric amyloid beta are covered in chapters 4-6. Despite being ultimately unsuccessful, this part provided a deeper understanding of the caveats of DNA-based assembly of immunogens and was instrumental for the preparation of the second generation of DNA-assembled vaccines directed against SARS-CoV2. We demonstrated that these vaccines, despite their simplicity, can produce broadly neutralizing immune responses.

We concluded that DNA-based assembly could be used for the preparation of effective modular vaccines, but their efficacy is contingent upon the biological stability of the DNA duplexes. Formulation via attachment to gold nanoparticles was used as such a strategy, and the generated immune responses demonstrated the efficient formation of immunological synapses and activation of B cells. Alternative strategies for the stabilization of the assemblies in the absence of gold are discussed.

Finally, we studied DNA nanorings and the mechanical forces which come to exist in them due to the different stiffness of double- and single-stranded DNA. We developed a model for the generation of mechanically stressed ssDNA/dsDNA hybrid nanorings and characterized them using atomic force microscopy.

Comments

2022 Copyright, the authors

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