ExPEC glycoconjugate vaccine formulations
Compositions and methods for inducing an immune response against extra-intestinal pathogenic Escherichia coli (ExPEC) are described. In particular, multivalent vaccines containing O-antigen polysaccharide covalently bound to an exotoxin A of Pseudomonas aeruginosa (EPA) carrier protein that can withstand multiple environmental stresses are described.
1. A composition comprising:
(a) at least one O-antigen polysaccharide, wherein the at least one O-antigen polysaccharide is covalently bound to an exotoxin A of Pseudomonas aeruginosa (EPA) carrier protein;
(b) 3% to 8% (w/v) sorbitol;
(c) 5 to 15 mM methionine;
(d) 5 to 20 mM phosphate buffer comprising potassium phosphate and sodium phosphate at a pH of 6.5 to 7.5;
(e) 0.01% to 0.2% (w/v) surfactant; and
(f) water.
2. The composition of claim 1 , wherein the concentration of sorbitol is 5% (w/v).
3. The composition of claim 1 , wherein the concentration of methionine is 10 mM.
4. The composition of claim 1 , wherein the concentration of the phosphate buffer is 10 mM, and the pH of the phosphate buffer is 7.0.
5. The composition of claim 1 , wherein the surfactant is polysorbate 80 (PS80).
6. The composition of claim 1 , wherein the surfactant is selected from the group consisting of poloxamer 188, polysorbate 20 (PS20), and polysorbate 80 (PS80).
7. The composition of claim 1 , which is stable for at least 6 months when stored at a temperature of 2-8° C.
8. The composition of claim 1 , comprising two or more O-antigen polysaccharides, wherein the concentration of each of the O-antigen polysaccharides is independently about 1 to 200 μg/mL.
9. The composition of claim 1 , wherein the polysaccharide:carrier protein weight/weight ratio is 1:10 to 1:2.
10. The composition of claim 1 , which is stable for at least 6 months when stored at a temperature of −18° C. to −80° C.
11. The composition of claim 1 , comprising a concentration of total O-antigen polysaccharide of 4 to 1000 μg/mL.
12. The composition of claim 1 , comprising a total concentration of EPA carrier protein of 40 to 2000 μg/mL.
13. The composition of claim 1 , in a container made of glass, polycarbonate, or polyethylene terephthalate.
14. A composition comprising:
(a) at least one O-antigen polysaccharide, wherein the at least one O-antigen polysaccharide is covalently bound to an exotoxin A of Pseudomonas aeruginosa (EPA) carrier protein;
(b) 5% (w/v) sorbitol;
(c) 10 mM methionine;
(d) 10 mM phosphate buffer comprising KH 2 PO 4 and Na 2 HPO 4 at a pH of 7.0;
(e) 0.02% (w/v) polysorbate 80 (PS80); and
(f) water.
15. A method of preparing a composition according to claim 1 , the method comprising mixing:
(a) at least one O-antigen polysaccharide, wherein the at least one O-antigen polysaccharide is covalently bound to an exotoxin A of Pseudomonas aeruginosa (EPA) carrier protein;
(b 3% to 8% (w/v) sorbitol;
(c) 5 to 15 mM methionine;
(d) 5 to 20 mM phosphate buffer comprising potassium phosphate and sodium phosphate at a pH of 6.5 to 7.5;
(e) 0.01% to 0.2% (w/v) surfactant; and
(f) water,
to thereby prepare the composition.
16. The method according to claim 15 , further comprising storing the composition at a temperature of 2-8° C. for at least 6 months.
17. The method according to claim 15 , further comprising storing the composition at a temperature of −18° C. to −80° C. for at least 6 months.
18. A method for stably maintaining an O-antigen polysaccharide in a liquid composition, comprising preparing the liquid composition and storing the liquid composition at a temperature of 2-8° C., wherein the liquid composition comprises:
(a) 1 to 200 μg/mL of the O-antigen polysaccharide covalently bound to an exotoxin A of Pseudomonas aeruginosa (EPA) carrier protein;
(b) 3% to 8% (w/v) sorbitol;
(c) 5 to 15 mM methionine;
(d) 5 to 20 mM phosphate buffer comprising potassium phosphate and sodium phosphate at a pH of 6.5 to 7.5;
(e) 0.01% to 0.2% (w/v) surfactant, and
(f) water.
19. The method of claim 18 , wherein the liquid composition comprises:
(a) 1 to 200 μg/mL of the O-antigen polysaccharide covalently bound to the EPA carrier protein, optionally one or more other O-antigen polysaccharides covalently bound to an EPA carrier protein;
(b) 5% (w/v) sorbitol;
(c) 10 mM methionine;
(d) 10 mM phosphate buffer comprising KH 2 PO 4 and Na 2 HPO 4 at a pH of 7.0;
(e) 0.02% (w/v) polysorbate 80 (PS80); and
(f) water.
20. The composition of claim 1 , in a syringe.