Vaccine compositions having improved stability and immunogenicity
Disclosed herein are nanoparticles suitable for use in vaccines. The nanoparticles present antigens from pathogens surrounded to and associated with a detergent core resulting in enhanced stability and good immunogenicity. Dosages, formulations, and methods for preparing the vaccines and nanoparticles are also disclosed.
1. A composition comprising:
(i) a nanoparticle comprising a non-ionic detergent core and a viral glycoprotein, wherein the viral glycoprotein consists of a trimeric influenza HA glycoprotein, wherein the influenza HA glycoprotein contains a head region that projects outward from the non-ionic detergent core and a transmembrane domain that is associated with the non-ionic detergent core, wherein the amino acid sequence of the influenza HA glycoprotein has 100% identity to the amino acid sequence of the native influenza HA protein; wherein the non-ionic detergent is PS80;
and
(ii) a pharmaceutically acceptable buffer.
2. The composition of claim 1 , wherein the influenza HA protein sub-type is selected from the group consisting of H1, H3, H4, H5, and H7.
3. The composition of claim 1 , wherein the pharmaceutically acceptable buffer comprises (i) sodium phosphate at 15 mM to 50 mM; (ii) NaCl at about 150 to about 250 mM; wherein the composition pH is between 7.0 and 7.5.
4. The composition of claim 1 , wherein the pharmaceutically acceptable buffer comprises (i) sodium phosphate at about 25 mM; (ii) NaCl at about 150 mM; wherein the composition pH is about 7.5.
5. A vaccine composition comprising:
(i) a nanoparticle comprising a non-ionic detergent core and a viral glycoprotein, wherein the viral glycoprotein consists of a trimeric influenza HA glycoprotein, and wherein the non-ionic detergent is PS80, wherein the influenza HA glycoprotein contains a head region that projects outward from the non-ionic detergent core and a transmembrane domain that is associated with the non-ionic detergent core, wherein the amino acid sequence of the influenza HA glycoprotein has 100% identity to the amino acid sequence of the native influenza HA protein;
(ii) a buffer comprising 25 mM sodium phosphate, pH 7.5, and 150 mM sodium chloride; and
(iii) an ISCOM matrix adjuvant.
6. The vaccine composition of claim 5 , wherein the ISCOM matrix adjuvant comprises a first ISCOM matrix containing fraction A, and a second ISCOM matrix containing fraction C.
7. The vaccine composition of claim 6 , wherein the first ISCOM matrix comprises 70% to 95% by weight of the total ISCOM matrix, and the second ISCOM matrix comprises the remainder.
8. The vaccine composition of claim 7 , wherein the first ISCOM matrix comprises 85% by weight of the total ISCOM matrix, and the second ISCOM matrix comprises the remainder.
9. The vaccine composition of claim 5 , wherein the influenza HA protein sub-type is selected from the group consisting of H1, H3, H4, H5, and H7.
10. A method of preparing a recombinant influenza nanoparticle comprising steps of:
(i) binding a protein extract comprising a first detergent and influenza HA glycoprotein to a protein purification column, wherein the column binds the influenza glycoprotein;
(ii) performing a detergent exchange by substantially replacing the first detergent with a second detergent; and
(iii) eluting the bound influenza glycoprotein from the column in the presence of the second detergent to provide the nanoparticle, wherein the second detergent is PS80;
wherein the transition midpoint (Tm) of the nanoparticle, as measured by differential scanning calorimetry is at least about 60, wherein no buffer used during preparation has a pH of below 7.0, and wherein the amino acid sequence of the influenza HA glycoprotein has 100% identity to the amino acid sequence of the native influenza HA glycoprotein, and wherein the nanoparticle consists of a viral glycoprotein and a non-ionic detergent core, wherein the viral glycoprotein is a trimeric HA glycoprotein, and wherein the HA glycoprotein contains a head region that projects outward from the non-ionic detergent core and a transmembrane domain that is associated with the non-ionic detergent core, and wherein the non-ionic detergent is PS80.
11. The method of claim 10 , wherein the first detergent is NP-9.
12. The composition of claim 1 , wherein the nanoparticle has a transition midpoint (Tm) peak in a range of about 60° C. to about 75° C. and wherein the Tm is measured by Differential Scanning calorimetry.
13. The vaccine composition of claim 5 , wherein the nanoparticle has a transition midpoint (Tm) peak in a range of about 60° C. to about 75° C. and wherein the Tm is measured by Differential Scanning calorimetry.
14. The composition of claim 1 , further comprising a second nanoparticle comprising a second influenza HA glycoprotein, wherein the HA glycoprotein contains a head region, which projects outward from the non-ionic detergent core, and a transmembrane domain, which is associated with the non-ionic detergent core, wherein the second influenza HA glycoprotein comprises an HAI polypeptide and an HA2 polypeptide connected by a disulfide bond, wherein the non-ionic detergent is PS80.
15. The vaccine composition of claim 5 , further comprising a second nanoparticle comprising a second influenza HA glycoprotein, wherein the HA glycoprotein contains a head region, which projects outward from the non-ionic detergent core, and a transmembrane domain, which is associated with the non-ionic detergent core, wherein the second influenza HA glycoprotein comprises an HAI polypeptide and an HA2 polypeptide connected by a disulfide bond, wherein the non-ionic detergent is PS80.
16. The composition of claim 1 , comprising a second nanoparticle, wherein the HA glycoprotein in the first nanoparticle is from a different influenza strain than the second nanoparticle.
17. The composition of claim 1 , comprising three nanoparticles, wherein the HA glycoprotein in each nanoparticle is from a different influenza strain than the other two nanoparticles.
18. The composition of claim 1 , comprising four nanoparticles, wherein the HA glycoprotein in each nanoparticle is from a different influenza strain than the other three nanoparticles.
19. The vaccine composition of claim 5 , comprising a second nanoparticle, wherein the HA glycoprotein in the first nanoparticle is from a different influenza strain than the second nanoparticle.
20. The vaccine composition of claim 5 , comprising three nanoparticles, wherein the HA glycoprotein in each nanoparticle is from a different influenza strain than the other two nanoparticles.
21. The vaccine composition of claim 5 , comprising four nanoparticles, wherein the HA glycoprotein in each nanoparticle is from a different influenza strain than the other three nanoparticles.
22. The composition of claim 1 , wherein the HA glycoprotein is a HA0 glycoprotein.
23. The vaccine composition of claim 5 , wherein the HA glycoprotein is a HA0 glycoprotein.
24. The method of claim 10 , wherein the HA glycoprotein is a HA0 glycoprotein.