Peptides and methods for the detection of lyme disease antibodies
The invention provides compositions (e.g., peptide compositions) useful for the detection of antibodies that bind to Borrelia antigens. The peptide compositions comprise polypeptide sequences comprising variants in the IR6 domain of the Borrelia VlsE protein. The invention also provides devices, methods, and kits comprising such peptide compositions and useful for the detection of antibodies that bind to Borrelia antigens and the diagnosis of Lyme disease.
1. A mixture of isolated peptides comprising three or more different isolated peptides, wherein each isolated peptide comprises a sequence of L-K-K-D-D-N-I-A-A-A-X 11 -V-L-R-G-X 16 -X 17 -K-D-G-X 21 -F-A-X 24 -X 25 (SEQ ID NO: 1) wherein X 11 is an amino acid selected from the group consisting of V and L, X 16 is an amino acid selected from the group consisting of L and I, X 17 is an amino acid selected from the group consisting of A and V, X 21 is an amino acid selected from the group consisting of R, D and N, X 24 is an amino acid selected from the group consisting of I, W, and Y, and X 25 is an amino acid selected from the group consisting of K and R.
2. The mixture of claim 1 , wherein at least one of the isolated peptides comprises an additional N-terminal peptide sequence, wherein the additional N-terminal peptide sequence is a native VlsE sequence or non-VlsE Borrelia antigen.
3. The mixture of claim 2 , wherein the additional N-terminal peptide sequence is a sequence of n 1 -n 2 -S-P-n 5 -n 6 -P (SEQ ID NO: 149) or a fragment thereof, wherein n 1 is an amino acid selected from the group consisting of A and V, n 2 is an amino acid selected from the group consisting of E and D, n 5 is an amino acid selected from the group consisting of K and R, and n 6 is an amino acid selected from the group consisting of K and R.
4. The mixture of claim 1 , wherein at least one of the isolated peptides comprises an additional C-terminal peptide sequence, wherein the additional C-terminal peptide sequence is a native VlsE sequence or non-VlsE Borrelia antigen.
5. The mixture of claim 4 , wherein the additional C-terminal peptide sequence is a sequence of V-c 2 -E-G-c 5 -Q-Q-E-G-A-Q-Q-P-S-C (SEQ ID NO: 150) or a fragment thereof, wherein c 2 is an amino acid selected from the group consisting of Q and R, and c 5 is an amino acid selected from the group consisting of V and A.
6. The mixture of claim 4 , wherein the additional C-terminal peptide sequence is a sequence of A-V-c 3 -E-G-c 6 -Q-Q-E-G-A-Q-Q-P-S (SEQ ID NO: 151) or a fragment thereof, wherein c 3 is an amino acid selected from the group consisting of Q and R, and c 6 is an amino acid selected from the group consisting of V and A.
7. The mixture of claim 1 , wherein at least one of the isolated peptides comprises at least 30, 35, 40 or 45 amino acids.
8. The mixture of claim 1 , wherein at least one of said isolated peptides is conjugated to a ligand.
9. The mixture of claim 1 , wherein at least one of said isolated peptides is biotinylated.
10. The mixture of claim 1 , wherein at least one of said isolated peptides is conjugated to streptavidin or bovine serum albumin optionally via an additional terminal amino acid linking residue.
11. The mixture of claim 1 , wherein at least one of said isolated peptides is attached or immobilized to a solid support.
12. The mixture of claim 1 , wherein at least one of said isolated peptides is attached or immobilized to a bead, a flow path in a lateral flow immunoassay device, a well in a microtiter plate, or a flow path in a rotor.
13. A method for diagnosing Lyme disease in a subject, the method comprising:
contacting a sample from the subject with the mixture of isolated peptides of claim 1 ; and
detecting formation of an antibody-peptide complex comprising at least one of said isolated peptides, wherein formation of the complex is indicative the subject having Lyme disease.