Methods for preparing vesicles and formulations produced therefrom
View Patent ↗The present disclosure provides methods for preparing vesicles. In general, these methods include steps of providing a lyophilized lipid product and rehydrating the lyophilized lipid product with an aqueous solution comprising an antigen such that antigen-containing vesicles are formed. The lyophilized lipid product is prepared by dissolving vesicle-forming lipids in a polar-protic water-miscible organic solvent to produce a lipid solution and then lyophilizing the lipid solution. The present disclosure also provides antigen-containing vesicle formulations prepared using these methods. The present disclosure also provides kits that include a lyophilized lipid product in a first container and an aqueous solution comprising an antigen in a second container.
1. A method of entrapping antigens within lipid vesicles, wherein the method comprises:
dissolving vesicle-forming lipids in tert-butanol to produce a lipid solution, wherein the lipid solution comprises 1-monopalmitoyl glycerol, dicetylphosphate, cholesterol, and a bile acid comprising sodium deoxycholate, at a ratio of 5:1:4:0.5;
lyophilizing the lipid solution to produce a lyophilized lipid product; and
rehydrating the lyophilized lipid product with an aqueous solution comprising an antigen consisting of an attenuated or inactivated virus such that antigen-containing lipid vesicles are formed.
2. The method of claim 1 , wherein the aqueous antigen solution further comprises a lyoprotectant.
3. The method of claim 2 , wherein the lyoprotectant is sucrose.
4. The method of claim 1 , wherein the attenuated virus or inactivated virus is hepatitis A or influenza.
5. The method of claim 1 , further comprising a step of adding an adjuvant after the antigen-containing lipid vesicles are formed.
6. The method of claim 1 , wherein the lyophilized lipid product comprises an adjuvant.
7. The method of claim 1 , further comprising a step of lyophilizing the antigen-containing lipid vesicles.
8. The method of claim 7 , further comprising a step of rehydrating the antigen-containing lipid vesicles after they have been lyophilized.
9. The method of claim 1 , wherein the antigen exhibits reduced antigenic integrity upon exposure to tert-butanol.