Peptides, devices, and methods for the detection of Ehrlichia antibodies

Patent metadata

Field	Value
Publication number	US-10948487-B2
Application number	US-201916552381-A
Country	US
Kind code	B2
Filing date	Aug 27, 2019
Priority date	Oct 11, 2012
Publication date	Mar 16, 2021
Grant date	Mar 16, 2021

Abstract

The invention provides peptide compositions and mixtures useful for the detection of antibodies that bind to Ehrlichia antigens. The peptide compositions and mixtures comprise polypeptide sequences based on an immunogenic fragment of the Ehrlichia Outer Membrane Protein 1 (OMP-1) protein. The invention also provides devices, methods, and kits comprising such peptide compositions and mixtures useful for the detection of antibodies that bind to Ehrlichia antigens and the diagnosis of monocytic and/or granulocytic ehrlichiosis.

First claim

What is claimed:

1. A population of isolated peptides comprising two or more different peptides, the population of peptides comprising:
(i) one or more peptides comprising the sequence of S-X 2 -K-E-X 5 -K-Q-X 8 -T-X 10 -X 11 -X 12 -X 13 -G-L-K-Q-X 18 -W-X 20 -G-X 22 -X 23 -X 24 -X 25 -X 26 -G-G-G-G-G-N-F-S-A-K-E-E-X 39 -A-E-T R-X 44 -T-F-G-L-X 49 -K-Q-Y-D-G-A-X 56 -I-X 58 -E-N-Q-V-Q-N-K-F-T-I-S-N-C(SEQ ID NO:72) or fragment thereof comprising at least 20 contiguous amino acids of SEQ ID NO: 72, wherein X 2 is an amino acid selected from the group consisting of A and V, X 5 is an amino acid selected from the group consisting of E and D, X 8 is an amino acid selected from the group consisting of T and P, X 10 is an amino acid selected from the group consisting of T and V, X 11 is an amino acid selected from the group consisting of G and A, X 12 is an amino acid selected from the group consisting of L and V, X 13 is an amino acid selected from the group consisting of Y and F, X 18 is an amino acid selected from the group consisting of D and N, X 20 is an amino acid selected from the group consisting of D and N, X 22 is an amino acid selected from the group consisting of S and V, X 23 is an amino acid selected from the group consisting of A, S, and T, X 24 is an amino acid selected from the group consisting of A and I, X 25 is an amino acid selected from the group consisting of T and P, X 26 is an amino acid selected from the group consisting of S, N, and K, X 39 is any amino acid, X 44 is any amino acid, X 49 is any amino acid, X 56 is any amino acid, and X 58 is any amino acid,
(ii) one or more peptides comprising the sequence of S-X 2 -K-E-X 5 -K-Q-X 8 -T-X 10 -X 11 -X 12 -X 13 -G-L-K-Q-X 18 -W-X 20 -G-X 22 -X 23 -X 24 -X 25 -X 26 -G-G-G-G-G-N-F-S-A-K-E-E-X 39 -A-X 41 -T-R-X 44 -T-F-G-X 48 -X 49 -K-Q-Y-D-G-A-X 56 -I-X 58 -E-N-Q-V-Q-N-K-F-T-I-S-N-C(SEQ ID NO: 3) or a fragment thereof comprising at least 20 contiguous amino acids of SEQ ID NO: 3, wherein X 2 is an amino acid selected from the group consisting of A and V, X 5 is an amino acid selected from the group consisting of E and D, X 7 is any amino acid, X 8 is an amino acid selected from the group consisting of T and P, X 10 is an amino acid selected from the group consisting of T and V, X 11 is an amino acid selected from the group consisting of G and A, X 12 is an amino acid selected from the group consisting of L and V, X 13 is an amino acid selected from the group consisting of Y and F, X 18 is an amino acid selected from the group consisting of D and N, X 20 is an amino acid selected from the group consisting of D and N, X 22 is an amino acid selected from the group consisting of S and V, X 23 is an amino acid selected from the group consisting of A, S, and T, X 24 in SEQ ID NOs: 72 and is an amino acid selected from the group consisting of A and I, X 25 is an amino acid selected from the group consisting of T and P, X 26 is an amino acid selected from the group consisting of S, N, and K, X 39 is any amino acid, X 41 is an amino acid selected from the group consisting of D and N, X 44 is any amino acid, X 48 is an amino acid selected from the group consisting of V and A, X 49 is any amino acid, X 56 is any amino acid, and X 58 is any amino acid,
(iii) one or more peptides comprising the sequence of F-S-A-K-E-E-X 7 -A-E-T-R-X 12 -T-F-G-L-X 17 -K-Q-Y-D-G-A-X 24 -I-X 26 -E-N-Q-V-Q-N-K-F-T-I-S-N-C(SEQ ID NO: 71) or a fragment thereof comprising at least 20 contiguous amino acids of SEQ ID NO: 71, wherein X 7 is any amino acid, X 12 is any amino acid, X 17 is any amino acid, X 24 is any amino acid, and X 26 is any amino acid, or
(iv) one or more peptides from each of (i) and (ii), one or more peptides from each of (i) and (iii), one or more peptides from each of (ii) and (iii), or one or more peptides from each of (i), (ii), and (iii).

2. The population of isolated peptides of claim 1 , wherein the population comprises a peptide according to (i), wherein X 39 of SEQ ID NO: 72 is K; X 44 of SEQ ID NO: 72 is selected from the group consisting of K and R; X 49 of SEQ ID NO: 72 is selected from the group consisting of E and D; X 56 of SEQ ID NO: 72 is selected from the group consisting of K and Q; and X 58 of SEQ ID NO: 72 is selected from the group consisting of E and T.

3. The population of isolated peptides of claim 1 , wherein the population comprises a peptide according to (ii), wherein X 39 of SEQ ID NO: 3 is K; X 44 of SEQ ID NO: 3 is selected from the group consisting of K and R; X 49 of SEQ ID NO: 3 is selected from the group consisting of E and D; X 56 of SEQ ID NO: 3 is selected from the group consisting of K and Q; and X 58 of SEQ ID NO: 3 is selected from the group consisting of E and T.

4. The population of isolated peptides of claim 1 , wherein the population comprises a peptide according to (iii), wherein X 7 of SEQ ID NO: 71 is K; X 12 of SEQ ID NO: 71 is selected from the group consisting of K and R; X 17 of SEQ ID NO: 71 is selected from the group consisting of E and D; X 24 of SEQ ID NO: 71 is selected from the group consisting of K and Q; and X 26 of SEQ ID NO: 71 is selected from the group consisting of E and T.

5. The population of isolated peptides of claim 1 , comprising three or more different peptides.

6. The population of isolated peptides of claim 1 , wherein the fragment comprising at least 20 contiguous amino acids of SEQ ID NO 72, 3, or 71 comprises a cysteine residue at the carboxyl terminus of the peptide.

7. The population of isolated peptides of claim 1 , wherein the fragment comprising at least 20 contiguous amino acids of SEQ ID NO 72, 3, or 71 further comprises a cysteine residue at the amino terminus of the peptide.

8. The population of isolated peptides of claim 1 , wherein the fragment comprises at least 25, 30, or 35 contiguous amino acids of SEQ ID NO: 72, 3, or 71.

9. The population of isolated peptides of claim 1 , wherein one or more peptides comprises an additional N-terminal peptide sequence that is a native OMP-1 sequence or non-OMP-1 Ehrlichia antigen.

10. The population of isolated peptides of claim 1 , wherein one or more peptides comprises an additional C-terminal peptide sequence that is a native OMP-1 sequence or non-OMP-1 Ehrlichia antigen.

11. The population of isolated peptides of claim 1 , wherein one or more peptides is conjugated to a ligand or is biotinylated.

12. The population of isolated peptides of claim 1 , wherein one or more peptides is conjugated to avidin, streptavidin, neutravidin, serum albumin, keyhole limpet hemocyanin (KLH), an enzyme, a metallic nanoparticle, or a nanoshell.

13. The population of isolated peptides of claim 1 , wherein one or more peptides is conjugated to bovine serum albumin.

14. The population of isolated peptides of claim 1 , comprising at least one peptide of (i), at least one peptide of (ii), and at least one peptide of (iii); wherein each peptide in the population is conjugated to bovine serum albumin.

15. The population of isolated peptides of claim 1 , wherein the peptides are immobilized to a solid support optionally through a metallic nanolayer.

16. The population of isolated peptides of claim 15 , wherein the solid support is a plurality of beads, a flow path in a lateral flow immunoassay device, a well in a microtiter plate, or a flow path in a rotor.

17. A population of isolated peptides comprising
(i) one or more peptides comprising the sequence of S-X 2 -K-E-X 5 -K-Q-X 8 -T-X 10 -X 11 -X 12 -X 13 -G-L-K-Q-X 18 -W-X 20 -G-X 22 -X 23 -X 24 -X 25 -X 26 -G-G-G-G-G-N-F-S-A-K-E-E

Assignees

• Zoetis Services Llc

Inventors

• Mehra Rajesh K
• Aron Kenneth P
• Bleile Dennis M
• Forsyth Timothy P
• Walker Jeremy D
• Cuesico Cristina R

Classifications

• G01N33/56911 Primary

  Bacteria · CPC title

• C07K17/14

  Peptides being immobilised on, or in, an inorganic carrier · CPC title

• G01N2469/20

  Detection of antibodies in sample from host which are directed against antigens from microorganisms · CPC title

• G01N2333/29

  from Richettsiales (o) · CPC title

• C07K14/29

  from Richettsiales (O) · CPC title