Functional influenza virus-like particles (VLPS)
The present invention discloses and claims virus like particles (VLPs) that express and/or contains seasonal influenza virus proteins, avian influenza virus proteins and/or influenza virus proteins from viruses with pandemic potential. The invention includes vector constructs comprising said proteins, cells comprising said constructs, formulations and vaccines comprising VLPs of the inventions. The invention also includes methods of making and administrating VLPs to vertebrates, including methods of inducing substantial immunity to either seasonal and avian influenza, or at least one symptom thereof.
1. A method of manufacturing a vaccine composition comprising
(a) expressing in a host cell at least one influenza VLP, wherein said VLP comprises influenza proteins M1, HA and NA,
(b) growing the host cell under conditions which allow the formation of VLPs,
(c) purifying the VLPs, and
(b) suspending the VLPs in a pharmaceutically acceptable carrier or excipient;
wherein the M1 protein is derived from the influenza strain A/Indonesia/5/05 and the HA and NA proteins are derived from a different influenza virus strain.
2. The method of claim 1 , wherein VLPs are produced expressing avian, pandemic, and/or seasonal influenza proteins.
3. The method of claim 1 , wherein the VLP comprises influenza proteins consisting of a HA protein and an NA protein from a single influenza strain, and an M1 protein from the influenza strain A/Indonesia/5/05.
4. The method of claim 1 , wherein the VLP comprises influenza proteins consisting of a HA protein from one influenza strain, an NA protein from a second influenza strain, and an M1 protein from the influenza strain A/Indonesia/5/05.
5. The method of claim 1 , wherein a second VLP comprising influenza proteins consisting of a HA protein and an NA protein from a second influenza strain, and an M1 protein from the influenza strain A/Indonesia/5/05, wherein the HA and NA proteins of the second VLP are from a different strain in the first VLP.
6. The method of claim 1 , wherein a third VLP comprising influenza proteins consisting of a HA protein and an NA protein from a third influenza strain, and an M1 protein from the influenza strain A/Indonesia/5/05, wherein the HA and NA proteins of the third VLP are from a different strain in the first and second VLPs.
7. The method of claim 1 , wherein a fourth VLP comprising influenza proteins consisting of a HA protein and an NA protein from a fourth influenza strain, and an M1 protein from the influenza strain A/Indonesia/5/05, wherein the HA and NA proteins of the fourth VLP are from a different strain in the first, second and third VLPs.
8. The method of claim 1 , wherein a recombinant construct encoding influenza proteins is used to transfect, infect, or transform the host cell.
9. The method of claim 1 , wherein the host cell is an Sf9 cell.
10. The method of claim 8 , wherein the recombinant construct is baculovirus.
11. The composition of claim 1 , wherein an immune response is stimulated against one or more influenza strains.
12. The method of claim 1 , wherein VLPs purification comprises one or more of filtration, chromatography, and centrifugation, or any combination thereof.
13. The method of claim 12 , wherein the purification comprises centrifugation and the centrifugation is gradient centrifugation.
14. The method of claim 12 , wherein the purification comprises chromatography and the chromatography is ion exchange chromatography.
15. The method of claim 1 , wherein a pharmaceutically acceptable carrier is selected from the group consisting of saline, buffered saline, dextrose, glycerol, and a sterile isotonic aqueous buffer, and combinations thereof.