Advisor: Julie Champion, Ph.D. (Chemical and Biomolecular Engineering)

Committee Members:

Jennifer E. Curtis, Ph.D. (Physics)

James E. Dahlman, Ph.D. (Biomedical Engineering)

Ravi S. Kane, Ph.D. (Chemical and Biomolecular Engineering)

Mark Prausnitz, Ph.D. (Chemical and Biomolecular Engineering) 

Surface Engineering of Protein Nanoparticles for Intranasal Vaccination

Intranasal delivery of vaccines offers a promising alternative approach to invasive intramuscular injection, with additional benefits such as inducing mucosal antibodies and cellular responses to neutralize pathogens before entering systemic circulation. However, nasal secretions and mucosa are biological barriers that have been shown to inhibit the delivery of antigens and nanoparticles to nasal-associated lymphoid tissue (NALT) and lungs. Protein nanoparticles are composed of proteins at high mass-to-carrier ratio, while allowing for biocompatibility and tunable physiochemical properties. They have been demonstrated to be effective vaccines and drug delivery carriers. The surfaces of these carriers can be decorated with coatings and chemical modifications, which can alter transport and immune responses due to their interaction with biological barriers and cells. In this work, we evaluate intranasal localization of engineered surface-coated protein nanoparticles and assess their immune response following vaccination in murine models. To understand the principles behind modifying nanoparticle surface formulations will assist in improving accessibility to the NALT and delivery of protein-based nanocarriers for non-vaccine intranasal delivery. We screened ovalbumin nanoparticles coated with polyethylene glycol (PEG) and layer-by-layer coating of trimethyl chitosan and CpG oligodeoxynucleotide adjuvants delivered intranasally in murine models and compared to unmodified protein nanoparticles. The localization and biodistribution were observed using non-invasive in vivo imaging and for regional localization and tissues using both flow cytometry and immunohistochemistry. Surface-coated nanoparticles were used for intranasal vaccination in a murine model and characterized for the mucosal antigen-specific response, as well as systemic humoral and cellular responses through antibody titers and T-cell activation. The findings and designs from screening coatings with model ovalbumin nanoparticles were incorporated into influenza antigen nanoparticle formulations.  Two influenza antigens (hemagglutinin and matrix protein 2 - (A/California/07/2009(H1N1)) were used to construct a subunit protein nanoparticle vaccine with surface structure control using bioconjugation. A layer-by-layer (LBL) coating approach was used to survey specific formulation based on their administration route. Overall, our findings indicated that LBL surface formulation improved nasal biodistribution and immune response upon intranasal delivery, highlighting a new nanoparticle formulation for nasal vaccines.