PROTEINASE K RESISTANT SURFACE PROTEIN OF NEISSERIA MENINGITIDIS

• US 20080124353A1
• Filed: 10/30/2007
• Published: 05/29/2008
• Est. Priority Date: 03/17/1995
• Status: Abandoned Application

• Current Assignee: ID Biomedical Corporation (GSK plc)
• Original Assignee: ID Biomedical Corporation (GSK plc)
• Inventors: Martin, Denis, Hamel, Josee, Brodeur, Bernard R., Rioux, Clement
• Application Number:
  US11/929,892
• Publication Number:
  US 20080124353A1
• Time in Patent Office: Days
• US Class Current: 424/190.1
• CPC Class Codes: A61K 2039/522 avirulent or attenuated
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: A61K 39/095 Neisseria
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/22 from Neisseriaceae (F)

Abstract

The identification of a highly conserved, immunologically accessible antigen at the surface of Neisseria facilitates treatment, prophylaxis, and diagnosis of Neisseria diseases. This antigen is highly resistant to Proteinase K and has an apparent molecular weight of 22 kDa on SDS-PAGE. Specific polynucleotides encoding proteins of this class have been isolated from three Neisseria meningitidis strains and from one Neisseria gonorrhoeae strain. These polynucleotides have been sequenced, and the corresponding full-length amino acid sequences of the encoded polypeptides have been deduced. Recombinant DNA methods for the production of the Neisseria surface protein, and antibodies that bind to this protein are also disclosed.

Claims

1. A method of isolating a polypeptide comprising:

a) isolating a culture of Neisseria meningitidis bacteria; and

b) isolating an outer membrane portion from the culture of the bacteria,

wherein the polypeptide comprises (i) an amino acid sequence at least 90% identical to the amino acid sequence set forth in SEQ ID NO;

2, SEQ ID NO;

4, or SEQ ID NO;

6, wherein the polypeptide is capable of eliciting an antibody that specifically binds to a protein consisting of the amino acid sequence set forth in SEQ ID NO;

2, SEQ ID NO;

4, or SEQ ID NO;

6;

or (ii) the amino acid sequence set forth in SEQ ID NO;

2, SEQ ID NO;

4, or SEQ ID NO;

6.

2. The method according to claim 1 further comprising isolating the polypeptide from the outer membrane portion.

3. The method of claim 1, further comprising treating the outer membrane portion with proteinase K.

4. The method of claim 1 wherein the polypeptide is substantially purified from other N. meningitidis proteins.

5. The method of claim 1 wherein the polypeptide is a recombinant polypeptide.

6. The method of claim 1 wherein the polypeptide is capable of inducing an immunological response to N. meningitidis.

7. The method of claim 1 wherein the polypeptide comprises a polypeptide fragment of the amino acid sequence set forth in any one of SEQ ID NO:

2, SEQ ID NO;

4, or SEQ ID NO;

6, wherein the polypeptide fragment has at least one immunogenic epitope, and wherein the polypeptide fragment is capable of eliciting an antibody that specifically binds to a protein consisting of the sequence set forth in any one of SEQ ID NO;

2, SEQ ID NO;

4, SEQ ID NO;

6, and SEQ ID NO;

8.

8. The method according to claim 7 wherein the polypeptide fragment comprises the amino acid sequence set forth at (a) residue 31 to residue 55 of SEQ ID NO:

2;

(b) residue 51 to residue 86 of SEQ ID NO;

2;

or (c) residue 110 to residue 140 of SEQ ID NO;

2.

9. The method according to claim 7 wherein the polypeptide fragment comprises the amino acid sequence set forth in any one of SEQ ID NO:

9, SEQ ID NO;

10, SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, SEQ ID NO;

14, SEQ ID NO;

15, SEQ ID NO;

16, SEQ ID NO;

17, SEQ ID NO;

18, SEQ ID NO;

19, SEQ ID NO;

20, SEQ ID NO;

21, SEQ ID NO;

22, SEQ ID NO;

23, SEQ ID NO;

24, SEQ ID NO;

25, and SEQ ID NO;

26.

10. A method of manufacturing a vaccine comprising (a) isolating the polypeptide according to the method of claim 1 or claim 7;

and (b) formulating the polypeptide with a pharmaceutically acceptable excipient.

11. The method according to claim 10 further formulating the polypeptide with a pharmaceutically acceptable adjuvant.

12. The method according to claim 10 wherein the vaccine is formulated in a suitable vehicle.

13. A method for producing a recombinant polypeptide wherein the recombinant polypeptide comprises (a) the amino acid sequence set forth from amino acid residue 31 to amino acid residue 55 of SEQ ID NO:

2;

(b) the amino acid sequence set forth from amino acid residue 51 to amino acid residue 86 of SEQ ID NO;

2;

or (c) the amino acid sequence set forth from amino acid residue 110 to amino acid residue 140 of SEQ ID NO;

2, said method comprising culturing a host cell that comprises an expression vector, wherein the expression vector comprises a polynucleotide that encodes the recombinant polypeptide, and wherein the polynucleotide is operatively linked to one or more expression control sequences.

14. The method according to claim 13 wherein the recombinant polypeptide at least 90% identical to the amino acid sequence set forth in SEQ ID NO:

2, SEQ ID NO;

4, SEQ ID NO;

6, or SEQ ID NO;

8.

15. The method according to claim 13 wherein the recombinant polypeptide is capable of inducing an immunological response against Neisseria.

16. The method according to claim 13 wherein the recombinant polypeptide is capable of eliciting an antibody that specifically binds to a protein consisting of the amino acid sequence set forth in SEQ ID NO:

2, SEQ ID NO;

4, SEQ ID NO;

6, or SEQ ID NO;

8.

17. The method according to claim 13 wherein the host cell is a eukaryotic cell or a bacteria cell.

18. The method according to claim 13 wherein the host cell is a bacterial cell.

19. The method according to claim 13 wherein the one or more expression control sequences is heterologous.

20. The method according to claim 18 further comprising (a) isolating a culture of the bacterial cell;

and (b) isolating an outer membrane portion from said bacterial cell culture.

21. The method according to claim 20 further comprising isolating the recombinant polypeptide from the outer membrane portion.

22. A method of manufacturing a vaccine comprising (a) producing a recombinant polypeptide according to the method of claim 13;

and (b) formulating the recombinant polypeptide with a pharmaceutically acceptable excipient.