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 Steptococcus pneumoniae serotypes.
1. A polysaccharide-carrier protein conjugate, wherein the polysaccharide comprises a repeating unit of the following structure:
wherein the carrier protein is CRM197; and further wherein the polysaccharide-carrier protein conjugate has a molecular weight from 1000 kDa to 10,000 kDa.
2. The polysaccharide-carrier protein conjugate of claim 1 , wherein the carrier protein is conjugated to the polysaccharide at the aldehyde of an activated repeating unit of the following structure:
3. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 50 kDa and 1,000 kDa.
4. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 100 kDa and 800 kDa.
5. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide has a molecular weight of between 100 kDa and 300 kDa.
6. The polysaccharide-carrier protein conjugate of claim 1 , wherein the degree of conjugation of the conjugate is between 2 and 15.
7. The polysaccharide-carrier protein conjugate of claim 1 , wherein the degree of conjugation of the conjugate is between 3 and 10.
8. The polysaccharide-carrier protein conjugate of claim 1 , wherein the degree of conjugation of the conjugate is between 5 and 10.
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 0.8 to 1.2.
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 0.94.
12. The polysaccharide-carrier protein conjugate of claim 1 , wherein the polysaccharide-carrier protein conjugate has a polysaccharide to carrier protein mass ratio of about 0.81.
13. 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.
14. 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.
15. 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.
16. 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.
17. The polysaccharide-carrier protein conjugate of claim 1 , wherein the degree of conjugation of the conjugate is between 5 and 10; wherein the mass ratio of polysaccharide to carrier protein in the conjugate is between 0.8 and 1.2; and wherein the conjugate comprises less than about 15% of free polysaccharide compared to the total amount of polysaccharide.
18. The polysaccharide-carrier protein conjugate of claim 2 , wherein the carrier protein is conjugated to the polysaccharide at the aldehyde in at least 50% of the repeating units.
19. The polysaccharide-carrier protein conjugate of claim 1 , wherein the carrier protein is conjugated to the polysaccharide at the aldehyde of an activated repeating unit of the following structure:
wherein the carrier protein is conjugated to the polysaccharide at the aldehyde in at least 50% of the repeating units; wherein the degree of conjugation of the conjugate is between 5 and 10; wherein the mass ratio of polysaccharide to carrier protein in the conjugate is between 0.8 and 1.2; and wherein the conjugate comprises less than about 15% of free polysaccharide compared to the total amount of polysaccharide.
20. An immunogenic composition comprising the polysaccharide carrier-protein conjugate of claim 1 , and a pharmaceutically acceptable carrier.
21. The immunogenic composition of claim 20 , further comprising one or more additional polysaccharide-carrier protein conjugates, wherein each of the conjugates comprises a polysaccharide of a particular S. pneumoniae serotype conjugated to CRM197, and wherein the S. pneumoniae serotypes are selected from the group consisting of: 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 7B, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 17F, 18C, 19A, 19F, 20, 21, 22A, 22F, 23A, 23B, 23F, 24F, 27, 28A, 31, 33F, 34, 35A, 35B, 35F, and 38.
22. The immunogenic composition of claim 21 , wherein the immunogenic composition is formulated to comprise: 0.4 to 4 μg/mL of each polysaccharide present, except for serotype 6B polysaccharide which, if present, comprises 0.8 to 8 μg/mL of polysaccharide; and CRM197 carrier protein in an amount from about 0.5× to 3× the total amount of polysaccharide.
23. The immunogenic composition of claim 22 , wherein the immunogenic composition is formulated to further comprise 150 mM sodium chloride, 20 mM L-histidine buffer, and 0.05 to 2% w/v surfactant.
24. The immunogenic composition of claim 23 , further comprising an adjuvant.
25. The immunogenic composition of claim 24 , wherein the adjuvant is an aluminum-based adjuvant.
26. The immunogenic composition of claim 25 , wherein the adjuvant is selected from the group consisting of aluminum phosphate, aluminum sulfate, and aluminum hydroxide.
27. The immunogenic composition of claim 26 , wherein the adjuvant is aluminum phosphate.
28. The immunogenic composition of claim 27 , wherein the aluminum phosphate adjuvant is present at a concentration from 0.05 to 0.5 mg/mL.
29. A method of inducing an immune response to a Streptococcus pneumonaie capsular polysaccharide, comprising administering to a human an immunologically effective amount of the immunogenic composition of claim 21 .
30. A method of inducing an immune response to a Streptococcus pneumonaie capsular polysaccharide, comprising administering to a human an immunologically effective amount of the immunogenic composition of claim 21 .
31. The method of claim 30 , wherein the immunogenic composition is a single 0.5 mL dose formulated to comprise: 2 μg of each polysaccharide present, except for serotype 6B polysaccharide which, if present, is 4 μg; about 32 μg CRM197 carrier protein; 0.125 mg aluminum phosphate adjuvant; 150 mM sodium chloride, 20 mM L-histidine buffer, and 0.2% w/v PS-20.
32. An immunogenic composition comprising the polysaccharide carrier-protein conjugate of claim 19 , and a pharmaceutically acceptable carrier.
33. The immunogenic composition of claim 32 , further comprising one or more additional polysaccharide-carrier protein conjugates, wherein each of the conjugates comprises a polysaccharide of a particular S. pneumoniae serotype conjugated to CRM197, and wherein the S. pneumoniae serotypes are selected from the group consisting of: 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 7B, 7C, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 17F, 18C, 19A, 19F, 20, 21, 22A, 22F, 23A, 23B, 23F, 24F, 27, 28A, 31, 33F, 34, 35A, 35B, 35F, and 38.
34. A method of inducing an immune response to a Streptococcus pneumonaie capsular polysaccharide, comprising administering to a human an immunologically effective amount of the immunogenic composition of claim 32 .
35. A method of inducing an immune response to a Streptococcus pneumonaie capsular polysaccharide, comprising administering to a human an immunologically effective amount of the immunogenic composition of claim 33 .
36. The method of claim 35 , wherein the immunogenic composition is a single 0.5 mL dose formulated to comprise: 2 μg of each polysaccharide present, except for serotype 6B polysaccharide which, if present, is 4 μg; about 32 μg CRM197 carrier protein; 0.125 mg aluminum phosphate adjuvant; 150 mM sodium chloride, 20 mM L-histidine buffer, and 0.2% w/v PS-20.