Compositions and Methods Related to Flavivirus Envelope Protein Domain III Antigens

• US 20080268423A1
• Filed: 08/18/2003
• Published: 10/30/2008
• Est. Priority Date: 08/16/2002
• Status: Active Grant

• Current Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Original Assignee: Board of Regents of the University of Texas System (University of Texas System)
• Inventors: Holbrook, Michael, Beasley, David, Barrett, Alan
• Granted Patent: US 7,785,799 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: G01N 2333/18 Togaviridae; Flaviviridae
• CPC Class Codes: G01N 2469/20 Detection of antibodies in sample from host which are directed against antigens from microorganisms
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention concerns methods and compositions involving flavivirus envelope protein domain III antigens for the detection of virus and detection of antibodies against the virus. Such methods and compositions may be used to detect TBE serocomplex viruses or West Nile virus infection in a subject, patient, animal or biological fluid. The present invention also concerns kits for implementing such methods. In some embodiments, kits contain a recombinant TBE serocomplex virus or West Nile virus envelope protein domain III antigen.

Claims

1. A method of screening for a flavivirus in a subject or animal host comprising:

a) contacting a sample from the subject or animal with a composition comprising a flavivirus envelope protein domain III polypeptide under conditions that permit formation of specific immunocomplex between an antibody in the sample and the envelope protein domain III polypeptide, wherein the envelope protein domain III polypeptide is a yellow fever virus envelope protein domain III polypeptide, West Nile virus envelope protein domain III polypeptide, St. Louis encephalitis virus envelope protein domain III polypeptide, Murray Valley encephalitis virus envelope protein domain III polypeptide, tick borne encephalitis serocomplex virus envelope protein domain III polypeptide or a combination thereof; and

b) detecting whether a specific immunocomplex is formed.

3. The method of claim 1, wherein the envelope protein domain III polypeptide is not a dengue fever virus envelope domain III polypeptide.

4. The method of claim 1, wherein the envelope protein domain III polypeptide is not a fusion protein.

5. The method of claim 1, wherein the envelope protein domain III polypeptide is a West Nile virus envelope protein domain III polypeptide.

6. The method of claim 1, wherein the envelope protein domain III polypeptide comprises an amino acid sequence that has at least an 80% identity with SEQ ID NO:

11.

10. The method of claim 1, wherein the envelope protein domain III polypeptide comprises amino acids 292 to 402 as set forth in SEQ ID NO:

3 or an amino acid sequence set forth in SEQ ID NO;

8-21.

11. The method of claim 1, wherein the envelope protein domain III polypeptide comprises an amino acid sequence as set forth in SEQ ID NO:

8-20.

12. The method of claim 1, further comprising at least a second envelope protein domain III polypeptide.

13. The method of claim 1, wherein the immunocomplex is detected using anti-antibody secondary reagents.

19. The method of claim 1, wherein the envelope protein domain III polypeptide is obtained from a bacteria, a mammalian or an insect cell comprising an expression vector encoding the envelope protein domain III polypeptide.

20. The method of claim 1, wherein the subject is infected with West Nile virus or a tick borne encephalitis serocomplex virus.

2. (canceled)

7-8. -8. (canceled)

14-18. -18. (canceled)

21. A composition comprising an isolated West Nile virus or tick borne encephalitis serocomplex virus envelope protein domain III polypeptide.

22. The composition of claim 21, wherein the West Nile virus envelope protein domain III polypeptide is derived from West Nile strain 382-99, EthAn4766, 385-99, Kunjin MRM16, Golblum, TL443, DakAnMg798, or 804994

23. The composition of claim 21, wherein the West Nile virus envelope protein domain III polypeptide comprises an amino acid sequence that is at least 80% identical to SEQ ID NO:

11.

28. The composition of claim 21, wherein the envelope protein domain III polypeptide is operatively linked to a substrate.

29. The composition of claim 28, wherein the substrate is a microtiter plate, a bead or a microarray.

30. The composition of claim 21, wherein the composition is a vaccine composition.

31. The composition of claim 30, further comprising an adjuvant.

24-25. -25. (canceled)

32. A kit for screening for flavivirus antibodies, in a suitable container means, comprising at least one envelope protein domain III polypeptide, wherein the at least one envelope protein domain III polypeptide is a yellow fever virus envelope protein domain III polypeptide, West Nile virus envelope protein domain III polypeptide, St. Louis encephalitis virus envelope protein domain III polypeptide, Murray Valley encephalitis virus envelope protein domain III polypeptide, a Central European encephalitis (CEE) virus envelope protein domain III polypeptide, a louping ill (LI) virus, a Russian spring-summer encephalitis (RSSE) virus envelope protein domain III polypeptide, a Langat (LGT) virus envelope protein domain III polypeptide, a Powassan virus (POW) envelope protein domain III polypeptide, an Alkhurma (ALK) envelope protein domain III polypeptide, a Kyasanur Forest disease (KFD) virus envelope protein domain III polypeptide, an Omsk hemorrhagic fever (OHF) virus envelope protein domain III polypeptide or a combination thereof.

43. A method of screening for flavivirus in a subject comprising:

a) contacting a sample from the subject with a composition from the kit of claim 32; and

,

b) detecting whether an immunocomplex is formed between an antibody and the at least one envelope protein domain III polypeptide.

33-40. -40. (canceled)

41. A kit for screening for West Nile virus antibodies or TBE serocomplex antibodies in a subject comprising:

a) an assay plate comprising a multiplicity of microtiter wells comprising a composition comprising at least one envelope protein domain III polypeptide capable of binding a flavivirus antibody in the sample that can specifically bind to at least one envelope protein domain III polypeptide; and

b) a container means comprising a labeled secondary antibody having specific binding affinity for a flavivirus antibody in the sample that can specifically bind to at least one envelope protein domain III polypeptide.

42. (canceled)