Chimeric immunogenic polypeptides

Patent metadata

Field	Value
Publication number	US-11780892-B2
Application number	US-202117331518-A
Country	US
Kind code	B2
Filing date	May 26, 2021
Priority date	Jul 27, 2018
Publication date	Oct 10, 2023
Grant date	Oct 10, 2023

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) immunogenic sequences TRP120 (SEQ ID NO:22) and TRP75 (SEQ ID NO:19), 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 TRP120 (SEQ ID NO: 22) and TRP75 (SEQ ID NO: 19), or a sequence 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 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 TRP140 (SEQ ID NO:23), A34N1 (SEQ ID NO:7), TRP63 (SEQ ID NO:18), TRP47 (SEQ ID NO:17), 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 TRP36, TRP140, TRP28, or HSP.

8. The polypeptide of claim 7 , wherein the TRP36R1 if present is repeated 2-6 times, and wherein the other immunogenic sequences are repeated 1-3 times.

9. The polypeptide of claim 8 , wherein the polypeptide comprises all of TRP32, TRP120, TRP36, TRP140, TRP28, and HSP.

10. The polypeptide of claim 9 , wherein the polypeptide comprises SEQ ID NO:28.

11. 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.

12. The polypeptide of claim 11 , wherein TRP120 and A34N1 are each contiguously repeated 1-3 times.

13. The polypeptide of claim 12 , wherein TRP120 and A34N1 are each contiguously repeated 2 times.

14. The polypeptide of claim 1 , wherein the polypeptide comprises A34N1, TRP47, and/or TRP63.

15. The polypeptide of claim 14 , wherein the polypeptide comprises A34N1.

16. The polypeptide of claim 1 , wherein the polypeptide comprises TRP36R1, TRP140, TRP95R, and/or TRP95C.

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

18. The polypeptide of claim 1 , wherein the isolated polypeptide comprises A34N1, TRP63, TRP47, TRP75, TRP28, TRP36, TRP19, TRP140, and/or HSP.

19. The polypeptide of claim 1 , wherein the polypeptide further comprises at least two of TRP36R1, TRP36R2, TRP36R3, and/or TRP36CO.

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

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

22. The polypeptide of claim 21 , wherein the linker is a glycine linker.

23. The polypeptide of claim 22 , wherein the glycine linker has the amino acid sequence -(G) X -, wherein X=3-5.

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

25. The pharmaceutical preparation of claim 24 , wherein the pharmaceutical preparation is formulated for parenteral, intravenous, subcutaneous, intranasal, sublingual, or intradermal administration.

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

27. The polypeptide of claim 26 , wherein the solid support is glass or plastic.

28. The polypeptide of claim 26 , wherein the solid support is comprised in a lateral flow assay, or microfluidic device.

29. An isolated polypeptide of Formula I (A s -B t —C u -D v -E w -F x -G y -H z ) n ,
wherein 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 TRP120 (SEQ ID NO:22) and TRP75 (SEQ ID NO:19).

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

31. 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.

32. 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.

33. A method of inducing an immune response in a mammalian subject comprising administering to the subject an effective amount of a pharmaceutical preparation comprising a polypeptide of a pharmaceutical composition of claim 30 .

34. 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 therapeutic compound to treat Ehrlichia infection in the subject.

35. The polypeptide of claim 1 , wherein the polypeptide comprises A34N1, TRP63, and TRP75.

36. The polypeptide of claim 35 , wherein the polypeptide comprises SEQ ID NO:34.

Assignees

• Res Found Dev

Inventors

• Mcbride Jere W
• Walker David H

Classifications

• C07K14/29 Primary

  from Richettsiales (O) · CPC title

• A61K39/0233 Primary

  Rickettsiales, e.g. Anaplasma · CPC title

• G01N33/56911

  Bacteria · 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