Immunogenic detoxified mutants of cholera toxin
View Patent ↗An immunogenic detoxified protein comprising the amino acid sequence of subunit A of a cholera toxin (CT-A) or a fragment thereof or the amino acid sequence of subunit A of an Escherichia coli heat labile toxin (LT-A) or a fragment thereof wherein the amino acids at, or in positions corresponding to Ser-63 and Arg-192 are replaced with another amino acid. The immunogenic detoxified protein is useful as vaccine for Vibrio cholerae or an enterotoxigenic strain of Escherichia coli and is produced by recombinant DNA means by site-directed mutagenesis.
1. An isolated immunogenic detoxified protein comprising the amino acid sequence of subunit A of a cholera toxin (CT-A) or the amino acid sequence of subunit A of an Escherichia coli heat labile toxin (LT-A) wherein the amino acids at positions corresponding to Ser-63 and Arg-192 of SEQ ID NO:7 or SEQ ID NO:8 are replaced with another amino acid, and further wherein the amino acid at position corresponding to Ser-63 is replaced with a lysine residue and the amino acid corresponding to position Arg-192 is replaced with a glycine or an asparagine residue.
2. A vaccine composition comprising an immunogenic detoxified protein according to claim 1 and a pharmaceutically acceptable carrier.
3. A vaccine composition according to claim 2 further comprising an adjuvant.
4. A vaccine composition according to claim 2 further comprising a second immunogenic antigen.
5. A method of vaccinating a mammal against Vibrio cholerae or an enterotoxigenic strain of Escherichia coli comprising administering an immunologically effective amount of an immunogenic detoxified protein according to claim 1 .
6. A process for the formulation of a vaccine comprising bringing an immunogenic detoxified protein according to claim 1 into association with a pharmaceutically acceptable carrier.
7. The process according to claim 6 , further comprising bringing the vaccine into association with an adjuvant.
8. The process according to claim 6 , further comprising bringing the vaccine into association with a second antigen.
9. The immunogenic detoxified protein of claim 1 , wherein the protein comprises the amino acid sequence of subunit A of a cholera toxin (CT-A).
10. The immunogenic detoxified protein of claim 9 , wherein the amino acid at, or in a position corresponding to, Arg-192 is replaced with Asn-192.
11. The immunogenic detoxified protein of claim 9 , wherein the amino acid at, or in a position corresponding to, Arg-192 is replaced with Gly-192.
12. The immunogenic detoxified protein of claim 1 , wherein the protein comprises the amino acid sequence of subunit A of an Escherichia coli heat labile toxin (LT-A).
13. The immunogenic detoxified protein of claim 2 , wherein the detoxified protein has a toxicity of less than 0.01% of the naturally occurring toxin counterpart.
14. The immunogenic detoxified protein of claim 9 , further comprising at least one CT subunit B.
15. The immunogenic detoxified protein of claim 12 , further comprising at least one LT subunit B.