Isolated polypeptide of the toxin A and toxin B proteins of
This present invention provides C-TAB.G5 and C-TAB.G5.1 isolated polypeptides comprising the receptor binding domains of C. difficile toxin A and toxin B as set forth in the amino acid sequences of SEQ ID NO: 2 and SEQ ID NO: 4. The C-TAB.G5 and C-TAB.G5.1 isolated polypeptides may be used to neutralize toxic effects of C. difficile toxin A and/or toxin B.
1. A method for producing a recombinant polypeptide derived from Clostridium difficile toxins A and B, the method comprising
i) introducing a recombinant nucleic acid encoding said polypeptide into a host cell, and
ii) culturing the host cell under conditions that allow expression of said polypeptide, wherein the polypeptide comprises an amino acid sequence having at least 85% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 4, wherein the sequence identity is determined by Needleman-Wunsch global alignment.
2. The method of claim 1 , wherein the polypeptide comprises 19 repeating units of the C-terminal domain of toxin A.
3. The method of claim 1 , wherein the polypeptide comprises at least 23 repeating units of the C-terminal domain of toxin B.
4. The method of claim 1 , wherein the polypeptide has the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4.
5. The method of claim 1 , wherein the polypeptide is capable of inducing neutralizing antibodies against both C. difficile toxins A and B.
6. The method of claim 1 , wherein the polypeptide is useful for the treatment and/or prevention of C. difficile associated disease (CDAD) or infection.
7. The method for producing the polypeptide of claim 1 , wherein the method further comprises
iii) isolating said polypeptide.
8. The method of claim 1 , wherein the host cell is E. coli.
9. A method for producing a pharmaceutical composition comprising a recombinant polypeptide derived from Clostridium difficile toxins A and B, the method comprising combining the recombinant polypeptide and a pharmaceutically acceptable carrier or excipient, wherein the polypeptide comprises an amino acid sequence having at least 85% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 4, wherein the sequence identity is determined by Needleman-Wunsch global alignment.
10. The method of claim 9 , wherein the polypeptide comprises 19 repeating units derived from the C-terminal domain of toxin A of C. difficile.
11. The method of claim 9 , wherein the polypeptide comprises at least 23 repeating units of the C-terminal domain of toxin B of C. difficile.
12. The method of claim 9 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4.
13. The method of claim 9 , wherein the composition is capable of inducing neutralizing antibodies against C. difficile toxins A and B.
14. A nucleic acid comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence having at least 85% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2 or SEQ ID NO: 4, wherein the sequence identity is determined by Needleman-Wunsch global alignment.
15. The nucleic acid of claim 14 , wherein the nucleic acid sequence encodes a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2 or SEQ ID NO: 4.
16. The nucleic acid of claim 14 , wherein the nucleic acid sequence encodes a polypeptide comprising the amino acid sequence having at least 99% sequence identity to the amino acid sequence as set forth in SEQ ID NO: 2 or SEQ ID NO: 4.
17. The nucleic acid of claim 14 , wherein the polypeptide comprises 19 repeating units derived from the C-terminal domain of toxin A of C. difficile and/or at least 23 repeating units of the C-terminal domain of toxin B of C. difficile.
18. The nucleic acid of claim 14 , wherein the nucleic acid sequence encodes a polypeptide consisting of the amino acid sequence as set forth in SEQ ID NO: 2.
19. The nucleic acid of claim 14 , wherein the nucleic acid sequence encodes a polypeptide consisting of the amino acid sequence as set forth in SEQ ID NO: 4.
20. A method for producing the nucleic acid of claim 14 , the method comprising:
i) amplifying a nucleic acid comprising the nucleic acid sequence; and/or
ii) introducing a vector comprising the nucleic acid sequence into a host cell, culturing the host cell under conditions that allow replication of the vector, and isolating the vector.