Pneumococcal polysaccharides and their use in immunogenic polysaccharide-carrier protein conjugates
The present invention provides capsular polysaccharides from Streptococcus pneumoniae serotypes identified using NMR. The present invention further provides polysaccharide-protein conjugates in which capsular polysaccharides from one or more of these serotypes are conjugated to a carrier protein such as CRM197. Polysaccharide-protein conjugates from one or more of these N serotypes may be included in multivalent pneumococcal conjugate vaccines having polysaccharides from multiple additional Steptococcus pneumoniae serotypes.
1. A polysaccharide-carrier protein conjugate wherein the polysaccharide comprises a repeating unit of the following structure:
wherein x indicates a molar ratio from 0 to 1.0; wherein the carrier protein is CRM197, diphtheria toxin fragment B (DTFB), DTFB C8, Diphtheria toxoid (DT), tetanus toxoid (TT), fragment C of TT, pertussis toxoid, cholera toxoid, E. coli LT, E. coli ST, or exotoxin A from Pseudomonas aeruginosa ; and further wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.5 to 2.0.
2. The polysaccharide-carrier protein conjugate of claim 1 , wherein the carrier protein is CRM197.
3. The polysaccharide-carrier protein conjugate of claim 1 , wherein the carrier protein is conjugated to the polysaccharide through the carbon 2 position of a glycerol-1-PO 4 sugar.
4. The polysaccharide-carrier protein conjugate of claim 2 , wherein the degree of conjugation of the conjugate is between 2 and 15.
5. The polysaccharide-carrier protein conjugate of claim 2 , wherein the degree of conjugation of the conjugate is between 3 and 13.
6. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.5 to 1.5.
7. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.8 to 1.2.
8. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate comprises less than about 50% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
9. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate comprises less than about 25% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
10. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate comprises less than about 20% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
11. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate comprises less than about 15% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
12. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 25 kDa and 5,000 kDa.
13. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 50 kDa and 1,000 kDa.
14. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 100 kDa and 300 kDa.
15. The polysaccharide-carrier protein conjugate of claim 2 , wherein the degree of conjugation of the conjugate is between 3-13; wherein the polysaccharide has a molecular weight of between 100 kDa and 300 kDa; wherein the mass ratio of polysaccharide to carrier protein in the conjugate is between 0.5 and 1.5; wherein the polysaccharide-carrier protein conjugate comprises less than about 15% of non-covalently associated polysaccharide compared to the total amount of polysaccharide; and wherein the polysaccharide-carrier protein conjugate has a molecular weight from 1,000 kDa to 10,000 kDa.
16. A polysaccharide-carrier protein conjugate wherein the polysaccharide comprises a repeating unit of the following structure:
wherein x indicates a molar ratio from 0 to 1.0; wherein the carrier protein is CRM197; and
further wherein the polysaccharide-carrier protein conjugate has a molecular weight from 1,000 kDa to 10,000 kDa.
17. The polysaccharide-carrier protein conjugate of claim 16 , wherein the carrier protein is conjugated to the polysaccharide through the carbon 2 position of a glycerol-1-PO 4 sugar.
18. The polysaccharide-carrier protein conjugate of claim 16 , wherein the degree of conjugation of the conjugate is between 2 and 15.
19. The polysaccharide-carrier protein conjugate of claim 16 , wherein the degree of conjugation of the conjugate is between 3 and 13.
20. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.5 to 2.0.
21. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.5 to 1.5.
22. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.8 to 1.2.
23. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide-carrier protein conjugate comprises less than about 50% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
24. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide-carrier protein conjugate comprises less than about 25% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
25. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide-carrier protein conjugate comprises less than about 20% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
26. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide-carrier protein conjugate comprises less than about 15% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
27. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide has a molecular weight of between 25 kDa and 5,000 kDa.
28. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide has a molecular weight of between 50 kDa and 1,000 kDa.
29. The polysaccharide-carrier protein conjugate of claim 16 , wherein the polysaccharide has a molecular weight of between 100 kDa and 300 kDa.