Multivalent pneumococcal vaccines

1. A multivalent vaccine comprising fifteen or more species of immunogenic complexes, wherein each species of the immunogenic complexes comprises:

(a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from a distinct Streptococcus pneumoniae serotype; and

(b) a fusion protein comprising:

(i) a biotin-binding moiety; and

(ii) at least one polypeptide antigen comprising:

a first polypeptide antigen comprising an SP1500 polypeptide or an antigenic fragment thereof; and

a second polypeptide antigen comprising an SP0785 polypeptide or an antigenic fragment thereof;

wherein the fusion protein comprises an amino acid sequence at least 90% identical to SEQ ID NO:6; and

wherein in each species of the immunogenic complexes, the biotinylated polysaccharide antigen is non-covalently associated with the biotin-binding moiety of the fusion protein.

2. The vaccine of claim 1 , wherein the Streptococcus pneumoniae serotype for each species of the immunogenic complexes is selected from 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 6E, 6F, 6G, 6H, 7A, 7B, 7C, 7F, 8, 9A, 9L, 9N, 9V, 10A, 10B, 10C, 10F, 11A, 11B, 11C, 11D, 11E, 11F, 12A, 12B, 12F, 13, 14, 15A, 15B, 15C, 15F, 16A, 16F, 17A, 17F, 18A, 18B, 18C, 18F, 19A, 19B, 19C, 19F, 20A, 20B, 21, 22A, 22F, 23A, 23B, 23F, 24A, 24B, 24F, 25A, 25F, 27, 28A, 28F, 29, 31, 32A, 32F, 33A, 33B, 33C, 33D, 33E, 33F, 34, 35A, 35B, 35C, 35F, 36, 37, 38, 39, 40, 41A, 41F, 42, 43, 44, 45, 46, 47A, 47F, and 48.

3. The vaccine of claim 1 , wherein the Streptococcus pneumoniae serotype for each species of the immunogenic complexes is selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20B, 22F, 23F, and 33F.

4. The vaccine of claim 1 , wherein the first polypeptide antigen comprises an amino acid sequence at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to SEQ ID NO:5 or an antigenic fragment thereof.

5. The vaccine of claim 1 , wherein the second polypeptide antigen comprises an amino acid sequence at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to SEQ ID NO:4 or an antigenic fragment thereof.

6. The vaccine of claim 1 , wherein the fusion protein comprises an amino acid sequence at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to SEQ ID NO:6.

7. The vaccine of claim 1 , wherein the biotin-binding moiety is a polypeptide comprising an amino acid sequence at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to SEQ ID NO:2 or a biotin-binding fragment thereof.

8. The vaccine of claim 1 , wherein the fusion protein comprises the amino acid sequence of SEQ ID NO: 6.

9. The vaccine of claim 2 , comprising twenty four or more species of the immunogenic complexes.

10. The vaccine of claim 9 , comprising:

a first immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 1, and (b) a fusion protein;

a second immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 2, and (b) a fusion protein;

a third immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 3, and (b) a fusion protein;

a fourth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 4, and (b) a fusion protein;

a fifth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 5, and (b) a fusion protein;

a sixth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 6A, and (b) a fusion protein;

a seventh immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 6B, and (b) a fusion protein;

an eighth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 7F, and (b) a fusion protein;

a ninth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 8, and (b) a fusion protein;

a tenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 9N, and (b) a fusion protein;

an eleventh immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 9V, and (b) a fusion protein;

a twelfth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 10A, and (b) a fusion protein;

a thirteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 11A, and (b) a fusion protein;

a fourteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 12F, and (b) a fusion protein;

a fifteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 14, and (b) a fusion protein;

a sixteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 15B, and (b) a fusion protein;

a seventeenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 17F, and (b) a fusion protein;

an eighteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 18C, and (b) a fusion protein;

a nineteenth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 19A, and (b) a fusion protein;

a twentieth immunogenic complex comprising (a) a biotinylated polysaccharide antigen, comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 19F, and (b) a fusion protein;

a twenty-first immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 20B, and (b) a fusion protein;

a twenty-second immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 22F, and (b) a fusion protein;

a twenty-third immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 23F and (b) a fusion protein; and

a twenty-fourth immunogenic complex comprising (a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from Streptococcus pneumoniae serotype 33F, and (b) a fusion protein;

wherein each fusion protein comprises:

(i) a biotin-binding moiety;

(ii) at least one polypeptide antigen comprising: a first polypeptide antigen comprising an amino acid sequence at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to SEQ ID NO:5 or an antigenic fragment thereof; and a second polypeptide antigen comprising an amino acid sequence at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to SEQ ID NO:4 or an antigenic fragment thereof;

and wherein in each of the immunogenic complexes, the biotinylated polysaccharide antigen is non-covalently associated with the biotin-binding moiety of the fusion protein.

11. A pharmaceutical composition comprising the vaccine of claim 1 , and a pharmaceutically acceptable carrier.

12. The pharmaceutical composition of claim 11 , further comprising one or more adjuvants.

13. The pharmaceutical composition of claim 12 , wherein the one or more adjuvants is or comprises a co-stimulation factor.

14. The pharmaceutical composition of claim 12 , wherein the one or more adjuvants are selected from the group consisting of aluminum phosphate, aluminum hydroxide, and phosphated aluminum hydroxide.

15. A method of making a multivalent vaccine, comprising mixing fifteen or more species of immunogenic complexes in a single formulation, wherein each species of the immunogenic complexes comprises:

(a) a biotinylated polysaccharide antigen comprising biotin and a polysaccharide antigen from a distinct Streptococcus pneumoniae serotype; and

(b) a fusion protein comprising:

(i) a biotin-binding moiety; and

(ii) at least one polypeptide antigen comprising one or more of:

a first polypeptide antigen comprising an SP1500 polypeptide or an antigenic fragment thereof; and

a second polypeptide antigen comprising an SP0785 polypeptide or an antigenic fragment thereof;

wherein the fusion protein comprises an amino acid sequence at least 90% identical to SEQ ID NO:6; and

wherein in each species of the immunogenic complexes, the biotinylated polysaccharide antigen is non-covalently associated with the biotin-binding moiety of the fusion protein.

16. The method of claim 15 , wherein the Streptococcus pneumoniae serotype for each species of the immunogenic complexes is selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20B, 22F, 23F, and 33F.