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 an influenza virus-like particle (VLP) comprising
(a) expressing in an Sf9 insect cell, one or more nucleic acids encoding an influenza
HA protein, and influenza NA protein, and an influenza M1 protein, wherein the M1 protein is derived from a different influenza virus strain as compared to the HA and NA proteins, and wherein the M1 protein is from the influenza strain A/Indonesia/5/05;
(b) growing the Sf9 insect cell under conditions which allow the formation of a VLP comprising the influenza proteins.
2. The method of claim 1 further comprising
(c) purifying the VLPs.
3. The method of claim 2 further comprising
(d) suspending the VLPs in a pharmaceutically acceptable carrier or excipient.
4. The method of claim 1 , wherein the nucleic acids are expressed in the Sf9 insect cell using a baculovirus construct.
5. The method of claim 1 , wherein VLPs are produced expressing avian, pandemic, and/or seasonal influenza proteins.
6. The method of claim 1 , wherein the HA protein and the NA protein are from the same influenza strain.
7. The method of claim 2 , wherein the purification comprises one or more of filtration, chromatography, and centrifugation.
8. The method of claim 7 , wherein the purification comprises centrifugation and the centrifugation is gradient centrifugation.
9. The method of claim 8 , wherein the purification comprises chromatography and the chromatography is ion exchange chromatography.
10. The method of claim 1 , wherein the pharmaceutically acceptable carrier comprises saline.
11. The method of claim 1 , wherein the M1 protein has the sequence of SEQ ID NO:49.