Chimeric immunogenic polypeptides

Patent metadata

Field	Value
Publication number	US-11046734-B2
Application number	US-201916524446-A
Country	US
Kind code	B2
Filing date	Jul 29, 2019
Priority date	Jul 27, 2018
Publication date	Jun 29, 2021
Grant date	Jun 29, 2021

Abstract

Provided herein are chimeric polypeptides that may be used, e.g., for the diagnosis of or vaccination against Ehrlichia chaffeensis and/or Ehrlichia canis.

First claim

What is claimed is:

1. An isolated polypeptide, wherein the isolated polypeptide comprises:
(i) TRP36R1 (SEQ ID NO:8) and TRP140 (SEQ ID NO: 23), or sequences at least 90% identical; and
(ii) wherein at least one of the immunogenic sequences is contiguously repeated in the polypeptide.

2. The isolated polypeptide of claim 1 , wherein each of the TRP36R1 (SEQ ID NO:8) and TRP140 (SEQ ID NO: 23), or sequences at least 90% identical are contiguously repeated 1, 2, 3, 4, 5, 6, or 7 times in the polypeptide.

3. The isolated polypeptide of claim 1 , wherein the isolated polypeptide further comprises one or more of (SEQ ID NOs:11-16 or 36-42), wherein the one or more of (SEQ ID NOs:11-16 or 36-42) are contiguously repeated 0, 1, 2, or 3 times.

4. The isolated polypeptide of claim 1 , wherein each of the immunogenic sequences are contiguously repeated from 1 to 3 times in the polypeptide.

5. The isolated polypeptide of claim 4 , wherein each of the immunogenic sequences are contiguously repeated from 1 to 2 times in the polypeptide.

6. The isolated peptide of claim 1 , wherein the isolated polypeptide comprises at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all of the following immunogenic sequences: TRP120 (SEQ ID NO:22), TRP140 (SEQ ID NO:23), A34N1 (SEQ ID NO:7), TRP63 (SEQ ID NO:18), TRP47 (SEQ ID NO:17), TRP75 (SEQ ID NO:19), TRP28 (SEQ ID NO:2), TRP36R1 (SEQ ID NO:8), TRP36R2 (SEQ ID NO:9), TRP36R3 (SEQ ID NO:10), TRP36CO (SEQ ID NO:36), TRP19 (SEQ ID NO:1), HSP (SEQ ID NO:24), or a sequence at least 90% identical; wherein each of the immunogenic sequences are contiguously repeated from 1 to 7 times in the polypeptide.

7. The polypeptide of claim 1 , wherein the isolated polypeptide comprises TRP36R1 and TRP140.

8. The polypeptide of claim 7 , wherein the TRP36R1 is contiguously repeated 4-8 times, and wherein the TRP140 is contiguously repeated 1-3 times.

9. The polypeptide of claim 8 , wherein the polypeptide comprises or consists of 8 repeats of TRP36R1 and 4 repeats of TRP140.

10. The polypeptide of claim 9 , wherein the polypeptide comprises or consists of SEQ ID NO:27.

11. The polypeptide of claim 7 , wherein the polypeptide further comprises TRP19.

12. The polypeptide of claim 11 , wherein the polypeptide comprises or consists of SEQ ID NO:26.

13. The polypeptide of claim 1 , wherein the isolated polypeptide comprises at least two, at least three, at least four, at least five or all of the immunogenic sequences: TRP32, TRP120, TRP36, TRP140, TRP28, and/or HSP.

14. The polypeptide of claim 13 , wherein the TRP36R1 is repeated 2-6 times, and wherein the other immunogenic sequences are repeated 1-3 times.

15. The polypeptide of claim 14 , wherein the polypeptide comprises all of TRP32, TRP120, TRP36, TRP140, TRP28, and HSP.

16. The polypeptide of claim 15 , wherein the polypeptide comprises or consists of SEQ ID NO:28.

17. The polypeptide of claim 13 , wherein the polypeptide comprises TRP120, TRP36, TRP140, and TRP28.

18. The polypeptide of claim 17 , wherein the polypeptide comprises or consists of SEQ ID NO:29.

19. The polypeptide of claim 1 , wherein the isolated polypeptide comprises at least three, at least four, at least five or all of TRP32R1, TRP32R2, TRP32R3, TRP32R4, TRP120, and A34N1.

20. The polypeptide of claim 19 , wherein TRP120 and A34N1 are each contiguously repeated 1-3 times.

21. The polypeptide of claim 20 , wherein TRP120 and A34N1 are each contiguously repeated 2 times.

22. The polypeptide of claim 21 , wherein the polypeptide comprises or consists of SEQ ID NO:25.

23. The polypeptide of claim 1 , wherein the polypeptide comprises a polypeptide of SEQ ID NOs: 25-35.

24. The polypeptide of claim 1 , wherein the different immunogenic sequences are not separated by a linker or a spacer.

25. The polypeptide of claim 1 , wherein the different immunogenic sequences are separated by a linker or a spacer.

26. The polypeptide of claim 25 , wherein the linker is a glycine linker.

27. The polypeptide of claim 26 , wherein the glycine linker has the amino acid sequence -(G) X -, wherein X=3-5.

28. The polypeptide of claim 1 , wherein the polypeptide is less than 500, less than 450, less than 400, less than 350, less than 300, less than 250, less than 200, or less than 150 amino acids in length.

29. The polypeptide of claim 1 , wherein the polypeptide is comprised in a pharmaceutical preparation.

30. The pharmaceutical preparation of claim 29 , wherein the pharmaceutical preparation is formulated for parenteral, intravenous, subcutaneous, intranasal, sublingual, or intradermal administration.

31. The polypeptide of claim 1 , wherein the polypeptide is attached to a solid support or is comprised in a diagnostic kit.

32. The polypeptide of claim 31 , wherein the solid support is glass or plastic.

33. The polypeptide of claim 31 , wherein the solid support is comprised in a lateral flow assay, or microfluidic device.

34. An isolated polypeptide of Formula I (A s -B t -C u -D v -E w -F x -G y -H z ) n ,
wherein each of A, B, C, D, E, F, G, and H is a peptide selected from SEQ ID NOs:1-24 and 36-42, or a sequence at least 90% identical to any one of (SEQ ID NOs:1-24 or 36-42),
wherein s, t, u, v, x, y, and z is an integer 0-8, wherein at least two of s-z are ≥1 and at least one of s-z is ≥2, and
wherein n is an integer 1-5;
wherein the polypeptide comprises TRP36R1 (SEQ ID NO:8) and TRP140 (SEQ ID NO: 23).

35. A pharmaceutical preparation comprising the polypeptide of claim 1 and a pharmaceutically acceptable excipient.

36. A method of detecting antibodies that specifically bind an Ehrlichia organism in a test sample, comprising: (a) contacting an isolated polypeptide of claim 1 ; (b) detecting the peptide-antibody complexes; wherein the detection of the peptide-antibody complexes is an indication that antibodies specific for an Ehrlichia organism are present in the test sample, and wherein the absence of the peptide-antibody complexes is an indication that antibodies specific an Ehrlichia organism are not present in the test sample.

37. A method of identifying an Ehrlichia infection in a mammalian subject comprising: (a) contacting a biological sample from the subject with an isolated polypeptide of claim 1 under conditions that allow peptide-antibody complexes to form; and (b) detecting the peptide-antibody complexes; wherein the detection of the peptide-antibody complexes is an indication that the subject has an Ehrlichia infection.

38. A method of inducing an immune response in a mammalian subject comprising administering to the subject an effective amount of a pharmaceutical preparation comprising the polypeptide of claim 1 .

39. A method of treating an Ehrlichia chaffeensis or Ehrlichia canis infection in a subject comprising:
(a) contacting a biological sample from the subject with an isolated polypeptide of claim 1 under conditions that allow peptide-antibody complexes to form;
(b) detecting the peptide-antibody complexes; wherein the detection of the peptide-antibody complexes is an indication that the subject has an Ehrlichia chaffeensis or Ehrlichia canis infection; and
(c) administering a therapeut

Assignees

• Res Found Dev

Inventors

• Mcbride Jere W
• Walker David H

Classifications

• C07K14/29 Primary

  from Richettsiales (O) · CPC title

• G01N33/6854

  Immunoglobulins · CPC title

• C07K2319/33

  fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies · CPC title

• A61K39/0233 Primary

  Rickettsiales, e.g. Anaplasma · CPC title

• G01N33/56911

  Bacteria · CPC title