Capsular gram-positive bacteria bioconjugate vaccines
The present invention encompasses a novel S. aureus bioconjugate vaccine. More generally, the invention is directed to Gram-positive and other bioconjugate vaccines containing a protein carrier, at least one polysaccharide such as a capsular Gram-positive polysaccharide, and, optionally, an adjuvant or pharmaceutically acceptable carrier. The instant invention also includes methods of producing Gram-positive and other bioconjugate vaccines. An N-glycosylated protein is also provided that contains one or more polysaccharides such as Gram-positive polysaccharides. The invention is additionally directed to engineered prokaryotic organisms comprising nucleotide sequences encoding a glycosyltransferase of a first prokaryotic organism and a glycosyltransferase of a second prokaryotic organism. The invention further includes plasmids and prokaryotic cells transformed with plasmids encoding polysaccharides and enzymes which produce an N-glycosylated protein and/or bioconjugate vaccine. Further, the invention is directed to methods of inducing an immune response in a mammal comprising administering said bioconjugate vaccines.
1. A Gram-negative host prokaryotic organism comprising:
(i) a nucleotide sequence encoding at least one glycosyltransferase from a Gram-positive bacterium, wherein said Gram-positive bacterium is a Serotype 5 isolate of Staphylococcus aureus ;
(ii) a nucleotide sequence encoding at least one glycosyltransferase from a Gram-negative bacterium;
(iii) a nucleotide sequence encoding a carrier protein, wherein said carrier protein is Pseudomonas aeruginosa Exotoxin, and wherein said carrier protein comprises the amino acid sequence D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline; and
(iv) a nucleotide sequence encoding an oligosaccharyl transferase.
2. The host prokaryotic organism of claim 1 , wherein said glycosyltransferase from a Gram-negative bacterium is a P. aeruginosa glycosyltransferase.
3. The host prokaryotic organism of claim 2 , wherein said S. aureus is a methicillin-resistant strain.
4. The host prokaryotic organism of claim 2 , comprising at least two glycosyltransferases from different Gram-positive bacterium strains.
5. The host prokaryotic organism of claim 2 , wherein said host organism is E. coli.
6. The host prokaryotic organism of claim 1 , wherein said host organism is E. coli.
7. The host prokaryotic organism of claim 3 , wherein said host organism is E. coli.
8. The host prokaryotic organism of claim 1 , wherein said oligosaccharyl transferase is from Campylobacter jejuni.
9. The host prokaryotic organism of claim 2 , wherein said oligosaccharyl transferase is from Campylobacter jejuni.
10. The host prokaryotic organism of claim 1 , wherein said carrier protein is engineered to comprise the amino acid sequence D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline.
11. The host prokaryotic organism of claim 2 , wherein said carrier protein is engineered to comprise the amino acid sequence D/E-X-N-Z-S/T, wherein X and Z may be any natural amino acid except proline.
12. A composition comprising the host prokaryotic organism of claim 1 .
13. A composition comprising the host prokaryotic organism of claim 2 .
14. A composition comprising the host prokaryotic organism of claim 6 .
15. A method of producing a bioconjugate comprising a S. aureus Serotype 5 isolate polysaccharide linked to P. aeruginosa Exotoxin, comprising culturing the host prokaryotic organism of claim 1 under conditions suitable for the production of proteins.
16. A method of producing a bioconjugate comprising a S. aureus Serotype 5 isolate polysaccharide linked to P. aeruginosa Exotoxin, comprising culturing the host prokaryotic organism of claim 2 under conditions suitable for the production of proteins.
17. A method of producing a bioconjugate comprising a S. aureus Serotype 5 isolate polysaccharide linked to P. aeruginosa Exotoxin, comprising culturing the host prokaryotic organism of claim 6 under conditions suitable for the production of proteins.