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Single-vial filovirus glycoprotein vaccines: Biophysical characteristics and immunogenicity after co-lyophilization
Kendall B Preston1, Teri Ann S Wong2, Albert To2
1Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, CO 80303, United States.
Abstract:
Zaire ebolavirus (EBOV), Sudan ebolavirus (SUDV), and Marburg marburgvirus (MARV) are the most prevalent and pathogenic species of filovirus. Previously, we showed that glycoprotein antigens from each virus could be lyophilized to create thermostable monovalent subunit vaccines. However, cross-protection is not expected from the monovalent vaccines and therefore developing a trivalent filovirus vaccine would be desirable. Subunit protein vaccines often require the addition of an adjuvant to sufficiently boost the immunogenicity. Typically, liquid suspensions or emulsions of adjuvants and lyophilized antigens are stored in separate vials to avoid destabilizing interactions and are only mixed immediately before administration. Herein, we describe the development and characterization of monovalent and trivalent filovirus vaccines that are co-lyophilized with a squalane-in-water emulsion adjuvant. We found that the single-vial presentation retained adjuvant particle diameter and zeta potential after lyophilization and reconstitution. Furthermore, the trivalent vaccines elicited high antibody levels against all three antigens in mice and non-human primates. These results advance the prospect of developing a single-vial trivalent filovirus vaccine, which would enable easier distribution and administration of the vaccine to resource-poor areas.
Insights
A new single-vial filovirus vaccine combines Zaire ebolavirus, Sudan ebolavirus, and Marburg marburgvirus antigens with an adjuvant. This co-lyophilized vaccine is thermostable and elicits strong immune responses, simplifying distribution.
Area of Science:
- Virology
- Immunology
- Vaccine Development
Background:
- Zaire ebolavirus (EBOV), Sudan ebolavirus (SUDV), and Marburg marburgvirus (MARV) are highly pathogenic filoviruses.
- Thermostable monovalent subunit vaccines against these viruses have been developed.
- A trivalent vaccine is desirable for broader protection, but subunit vaccines typically require adjuvants, complicating formulation.
Purpose of the Study:
- To develop and characterize a single-vial, co-lyophilized trivalent filovirus vaccine.
- To assess the stability of the vaccine formulation after lyophilization and reconstitution.
- To evaluate the immunogenicity of the trivalent vaccine in preclinical models.
Main Methods:
- Co-lyophilization of filovirus glycoprotein antigens (EBOV, SUDV, MARV) with a squalane-in-water emulsion adjuvant.
- Characterization of adjuvant particle size and zeta potential post-lyophilization and reconstitution.
- Immunization of mice and non-human primates with monovalent and trivalent vaccines.
- Measurement of antibody levels against all three filovirus antigens.
Main Results:
- The single-vial co-lyophilized formulation maintained adjuvant integrity (particle diameter and zeta potential).
- Trivalent vaccines successfully elicited high antibody titers against EBOV, SUDV, and MARV antigens.
- Preclinical models demonstrated the immunogenicity of the co-lyophilized trivalent vaccine.
Conclusions:
- A single-vial, co-lyophilized trivalent filovirus vaccine is feasible and immunogenic.
- This formulation simplifies vaccine preparation and administration.
- The development advances the potential for easier distribution of filovirus vaccines, particularly in resource-limited settings.
