Methods for conjugation of oligosaccharides or polysaccharides to protein carriers through oxime linkages via 3-deoxy-D-manno-octulsonic acid
Methods for preparing an oligosaccharide-protein carrier immunogenic conjugate or a polysaccharide-protein carrier immunogenic conjugate. The methods include obtaining an oligosaccharide or polysaccharide having a KDO moiety located at the terminal reducing end of the oligosaccharide or polysaccharide that includes a carbonyl functional group; and reacting the carbonyl functional group of the KDO moiety with an aminooxylated protein carrier molecule resulting in a conjugate that includes a covalent oxime bond between the oligosaccharide and the protein carrier or the polysaccharide and the protein carrier.
1. An immunogenic conjugate comprising the structure of:
Pr-Sp-O—N═C(COOH)-anh-KDO—OS
wherein Pr is a carrier protein, Sp is an optional spacer moiety, anh-KDO is an anhydro moiety from 3-deoxy-D-manno-octulosonic acid, and OS is an oligosaccharide or polysaccharide residue from the cleavage of Lipid A from a lipopolysaccharide, wherein the lipopolysaccharide is obtained from Haemophilus ducreyi, Bordetella bronchiseptica, Bordetetla parapertussis, Bordetella pertussis, Vibrio cholerae, Haemophilus influenza, or a mixture thereof.
2. The conjugate of claim 1 , wherein the oligosaccharide or polysaccharide includes an O-antigen chain and a core oligosaccharide.
3. The conjugate of claim 1 , wherein the lipopolysaccharide is obtained from Bordetella bronchiseptica or Bordetella parapertussis and the OS includes at least:
-4-β-ManNAc3ANcAN-4-β-GlcNAc3NAcAN-4-β-GalNAc-4-β-ManNAc3NAcA-3-β-FucNAc4NMe-.
4. A pharmaceutical composition comprising the conjugate of claim 3 and a pharmaceutically acceptable carrier.
5. The conjugate of claim 1 , wherein the lipopolysaccharide is obtained from Haemophilus ducreyi and the OS includes at least:
Hex 3 HexNAcHep 4 .
6. The conjugate of claim 1 , wherein the oligosaccharide or polysaccharide is obtained from Bordetella bronchiseptica, Bordetella parapertussis , or Bordetella pertussis.
7. The conjugate of claim 1 , wherein the oligosaccharide or polysaccharide is obtained from Bordetella bronchiseptica or Bordetella pertussis.
8. The conjugate of claim 1 , wherein the oligosaccharide or polysaccharide is obtained from Bordetella parapertussis.
9. A pharmaceutical composition comprising the conjugate of claim 1 and a pharmaceutically acceptable carrier.
10. The conjugate of claim 1 , wherein the lipopolysaccharide is obtained from Bordetella bronchiseptica.
11. A pharmaceutical composition comprising the conjugate of claim 10 and a pharmaceutically acceptable carrier.
12. The conjugate of claim 1 , wherein there are 5 to 20 OS chains bound to the single carrier protein.
13. The conjugate of claim 1 , wherein the OS is not oxidized.
14. A method of eliciting an immune response in a subject, comprising administering to the subject the conjugate of claim 1 , thereby eliciting an immune response in the subject.
15. An immunogenic conjugate comprising:
an oligosaccharide or polysaccharide having a core oligosaccharide, wherein the oligosaccharide or polysaccharide is obtained from Bordetella bronchiseptica, Bordetella parapertussis, Bordetella pertussis , or a mixture thereof; and
a carrier protein,
wherein the oligosaccharide or polysaccharide is conjugated to the carrier protein via an oxime covalent bond with a 3-deoxy-D-manno-octulsonic acid moiety located at the terminal reducing end of the oligosaccharide or polysaccharide.
16. The conjugate of claim 15 , wherein the oligosaccharide or polysaccharide is not oxidized.