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 serotypes may be included in multivalent pneumococcal conjugate vaccines having polysaccharides from multiple additional Streptococcus pneumoniae serotypes.
1. A polysaccharide-carrier protein conjugate, wherein the polysaccharide has an S. pneumoniae serotype 23B polysaccharide repeating unit of the following structure:
wherein the carrier protein is CRM197, and further wherein the conjugate has a molecular weight from 1,000 kDa to 10,000 kDa.
2. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.4 to 2.0.
3. The polysaccharide-carrier protein conjugate of claim 1 , wherein the carrier protein is conjugated to the serotype 23B polysaccharide unit through the 2 nd or 3 rd carbon of the rhamnose sugar.
4. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 50 kDa and 1,000 kDa.
5. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 100 kDa and 800 kDa.
6. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 100 kDa and 300 kDa.
7. The polysaccharide-carrier protein conjugate of claim 1 , wherein the degree of conjugation of the conjugate is between 2 and 15.
8. The polysaccharide-carrier protein conjugate of claim 1 , wherein the degree of conjugation of the conjugate is between 8 and 12.
9. 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 3.0.
10. 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.
11. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio of about 1.3.
12. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate comprises less than about 30% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
13. 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.
14. 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.
15. 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.
16. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 100 kDa and 300 kDa; wherein the degree of conjugation of the conjugate is between 4 and 7; wherein the mass ratio of polysaccharide to carrier protein in the conjugate is between 0.5 and 1.5; and wherein the conjugate comprises less than about 15% of free polysaccharide compared to the total amount of polysaccharide.
17. A polysaccharide-carrier protein conjugate, wherein the polysaccharide has an S. pneumoniae serotype 23B polysaccharide repeating unit of the following structure:
wherein the carrier protein is CRM197; and further wherein the wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.5 to 3.0.
18. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio from 0.5 to 1.5.
19. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio of about 1.3.
20. The polysaccharide-carrier protein conjugate of claim 17 , wherein the carrier protein is conjugated to the serotype 23B polysaccharide unit through the 2 nd or 3 rd carbon of the rhamnose sugar.
21. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide has a molecular weight of between 50 kDa and 1,000 kDa.
22. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide has a molecular weight of between 100 kDa and 800 kDa.
23. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide has a molecular weight of between 100 kDa and 300 kDa.
24. The polysaccharide-carrier protein conjugate of claim 17 , wherein the degree of conjugation of the conjugate is between 2 and 15.
25. The polysaccharide-carrier protein conjugate of claim 17 , wherein the degree of conjugation of the conjugate is between 8 and 12.
26. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide-carrier protein conjugate comprises less than about 30% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
27. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide-carrier protein conjugate comprises less than about 25% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
28. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide-carrier protein conjugate comprises less than about 20% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
29. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide-carrier protein conjugate comprises less than about 15% of non-covalently associated polysaccharide compared to the total amount of polysaccharide.
30. The polysaccharide-carrier protein conjugate of claim 17 , wherein the polysaccharide has a molecular weight of between 100 kDa and 300 kDa; wherein the degree of conjugation of the conjugate is between 4 and 7; wherein the mass ratio of polysaccharide to carrier protein in the conjugate is between 0.5 and 1.5; wherein the conjugate comprises less than about 15% of free polysaccharide compared to the total amount of polysaccharide; and wherein the conjugate has a molecular weight from 1,000 kDa to 10,000 kDa.