Poly-N-acetyl glucosamine (PNAG/dPNAG)-binding peptides and methods of use thereof
The present invention relates to peptides, particularly human monoclonal antibodies, that bind specifically to poly-N-acetyl glucosamine (PNAG), such as Staphylococcal PNAG, in acetylated, partially acetylated and/or fully deacetylated form. The invention further provides methods for using these peptides in the diagnosis, prophylaxis and therapy of infections by bacteria that express PNAG such as but not limited to Staphylococci and E. coli . Some antibodies of the invention enhance opsonophagocytic killing and in vivo protection against bacteria that express PNAG such as but not limited to Staphylococci and E. coli . Compositions of these peptides, including pharmaceutical compositions, are also provided, as are functionally equivalent variants of such peptides.
1. A composition comprising:
(i) a nucleic acid molecule having the nucleotide sequences of SEQ ID NO: 37, SEQ ID NO: 38, and SEQ ID NO: 39; and/or
(ii) a nucleic acid molecule having the nucleotide sequences of SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42;
wherein the nucleic acid molecule(s) of (i) and (ii) are contained in the same vector.
2. The composition of claim 1 , wherein the nucleotide sequences of SEQ ID NO: 37, SEQ ID NO: 38 and SEQ ID NO: 39 encode a heavy chain variable region of Staphylococcal poly-N-acetyl glucosamine/dPNAG-binding antibody (PNAG/dPNAG-binding antibody) and the nucleotide sequences of SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42 encode a light chain variable region of PNAG/dPNAG-binding antibody, the heavy chain variable region comprising a CDR1 having an amino acid sequence of SEQ ID NO: 13, a CDR2 having an amino acid sequence of SEQ ID NO: 14, and a CDR3 having an amino acid sequence of SEQ ID NO: 15, and the light chain variable region comprising a CDR1 having an amino acid sequence of SEQ ID NO: 16, a CDR2 having an amino acid sequence of SEQ ID NO: 17, and a CDR3 having an amino acid sequence of SEQ ID NO: 18.
3. The composition of claim 1 wherein the nucleic acid molecules are derived from a hybridoma having Accession No. PTA-5932.
4. The composition of claim 2 wherein the nucleic acid molecules are derived from a hybridoma having Accession No. PTA-5932.
5. The composition of claim 1 wherein the vector is a cloning vector or an expression vector.
6. A host cell transformed or transfected with the composition of claim 1 .
7. An expression vector comprising an isolated nucleic acid molecule having the nucleotide sequences of SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, operably linked to a promoter.
8. A host cell transformed or transfected with the expression vector of claim 7 .
9. A cloning vector comprising the isolated nucleic acid molecule having the nucleotide sequences of SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, operably linked to a promoter.
10. A host cell transformed or transfected with the cloning vector of claim 9 .
11. An isolated nucleic acid molecule comprising:
a heavy chain region comprising heavy chain CDR1, heavy chain CDR2 and heavy chain CDR3 sequences comprising nucleic acid sequences of SEQ ID NO: 37, SEQ ID NO: 38 and SEQ ID NO: 39, respectively; and
a light chain region comprising light chain CDR1, light chain CDR2 and light chain CDR3 sequences comprising amino acid sequences of SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively.
12. An expression vector comprising the isolated nucleic acid molecule of claim 11 , operably linked to a promoter.
13. A host cell transformed or transfected with the expression vector of claim 12 .
14. The composition of claim 1 wherein the nucleic acid molecules of (i) and (ii) are contained within an expression vector suitable for the recombinant expression of an antibody fragment selected from the group consisting of a F(ab′) 2 fragment, a Fab fragment, a Fv fragment, or a single-chain antibody; or wherein the nucleic acid molecules of (i) and (ii) are contained within an expression vector suitable for the recombinant expression of an antibody, or fragment thereof, having an IgG1, IgG3, IgG4, IgD, IgE, IgM, IgA1, IgA2, sIgA, or IgY isotype.
15. A recombinant nucleic acid molecule comprising one or both of the nucleotide sequences of SEQ ID NO: 38 and SEQ ID NO: 39, wherein the nucleic acid molecule comprises one or more restriction sites that are 5′ to, 3′ to, or both 5′ and 3′ to the one or both nucleotide sequences of SEQ ID NO: 38 and SEQ ID NO: 39.
16. The recombinant nucleic acid molecule of claim 15 further comprising one or more of the nucleotide sequences of SEQ ID NO: 37, SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42.
17. A recombinant nucleic acid molecule comprising one or more of the nucleotide sequences of SEQ ID NO: 40, SEQ ID NO: 41, and SEQ ID NO: 42, wherein the nucleic acid molecule comprises one or more restriction sites that are 5′ to, 3′ to, or both 5′ and 3′ to the one or more nucleotide sequences of SEQ ID NO: 40, SEQ ID NO: 41, and SEQ ID NO: 42.
18. The recombinant nucleic acid molecule of claim 17 further comprising one or more of the nucleotide sequences of SEQ ID NO: 37, SEQ ID NO: 38 and SEQ ID NO: 39.
19. A recombinant nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO: 37 and the nucleotide sequence of one or both SEQ ID NO: 38 and SEQ ID NO: 39, wherein the nucleic acid molecule comprises one or more restriction sites that are 5′ to, 3′ to, or both 5′ and 3′ to the nucleotide sequence of SEQ ID NO: 37.
20. The recombinant nucleic acid molecule of claim 15 further comprising one or more of the nucleotide sequences of SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42.
21. A recombinant nucleic acid molecule comprising
one or more of the nucleotide sequences of SEQ ID NO: 37, SEQ ID NO: 38, and SEQ ID NO: 39, wherein the nucleic acid molecule comprises one or more restriction sites that are 5′ to, 3′ to, or both 5′ and 3′ to the one or more nucleotide sequences of SEQ ID NO: 37, SEQ ID NO: 38, and SEQ ID NO: 39, and
one or more of the nucleotide sequences of SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42.