Complexes of cytomegalovirus proteins

Patent metadata

Field	Value
Publication number	US-9683022-B2
Application number	US-201314410461-A
Country	US
Kind code	B2
Filing date	Jun 29, 2013
Priority date	Jul 6, 2012
Publication date	Jun 20, 2017
Grant date	Jun 20, 2017

Abstract

An isolated human cytomegalovirus (HCMV) membrane protein complex that comprises gH, gL and at least one more HCMV glycoprotein is provided. In some embodiments the complex consists of gH, gL and gO. In other embodiments the complex consists of gH, gL, pUL128, pUL130 and pUL131A. Processes for expressing and purifying such complexes, and subsequent uses of such complexes in immunogenic compositions and vaccines, are also provided.

First claim

The invention claimed is:

1. A composition comprising a purified human Cytomegalovirus (HCMV) pentameric complex comprising: a pentamer-forming fragment of HCMV gH having a truncated transmembrane (TM) domain and a truncated ectodomain as compared to wild-type HCMV gH; HCMV gL or pentamer-forming fragment thereof; HCMV pUL128 or pentamer-forming fragment thereof; HCMV pUL130 or pentamer-forming fragment thereof; and HCMV pUL131A or pentamer-forming fragment thereof, wherein the truncated TM domain consists of an amino acid sequence having a deletion of the amino acids corresponding to residues 718 to 736 of wild-type sequence SEQ ID NO: 1, and wherein the truncated ectodomain consists of an amino acid sequence having a deletion of the amino acids corresponding to residues 716 and 717 of wild-type sequence SEQ ID NO: 1.

2. The composition of claim 1 , wherein said gL, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 7-9 and 31.

3. The composition of claim 1 , wherein said gL, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs: 7-9 and 31.

4. The composition of claim 1 , wherein said pUL128, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 13-15 and 33.

5. The composition of claim 1 , wherein said pUL128, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs: 13-15 and 33.

6. The composition of claim 1 , wherein said pUL130, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 16, 17, and 34.

7. The composition of claim 1 , wherein said pUL130, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs: 16, 17, and 34.

8. The composition of claim 1 , wherein said pUL131A, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 18-20, and 35.

9. The composition of claim 1 , wherein said pUL131A, or pentamer-forming fragment thereof, comprises a sequence selected from the group consisting of SEQ ID NOs: 18-20, and 35.

10. The composition of claim 1 , that is a pharmaceutical comprising the purified HCMV pentameric complex and an adjuvant.

11. The pharmaceutical of claim 10 , wherein the adjuvant comprises an aluminum salt, an oil-in-water emulsion, or a combination thereof.

12. The pharmaceutical of claim 11 , wherein the adjuvant comprising an aluminum salt comprises aluminum hydroxide, aluminum oxyhydroxide, aluminum hydroxide adsorbed to a TLR7 agonist, or a combination thereof.

13. The pharmaceutical of claim 11 , wherein the oil-in-water emulsion comprises MF59, AS03, or a combination thereof.

14. The composition of claim 1 , wherein said pentamer-forming fragment of HCMV gH comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 4, 6, 29, and 30.

15. The composition of claim 14 , wherein said pentamer-forming fragment of HCMV gH comprises a sequence selected from the group consisting of SEQ ID NOs: 4, 6, 29, and 30.

16. The composition of claim 15 , wherein said pentamer-forming fragment of HCMV gH comprises the sequence SEQ ID NO: 6.

17. A purified human Cytomegalovirus (HCMV) pentameric complex comprising: a pentamer-forming fragment of HCMV gH having a truncated transmembrane (TM) domain and a truncated ectodomain as compared to wild-type HCMV gH; HCMV gL or pentamer-forming fragment thereof; HCMV pUL128 or pentamer-forming fragment thereof; HCMV pUL130 or pentamer-forming fragment thereof; and HCMV pUL131A or pentamer-forming fragment thereof, wherein the truncated TM domain consists of an amino acid sequence having a deletion of the amino acids corresponding to residues 718 to 736 of wild-type sequence SEQ ID NO: 1, and wherein the truncated ectodomain consists of an amino acid sequence having a deletion of the amino acids corresponding to residues 716 and 717 of wild-type sequence SEQ ID NO: 1.

18. The purified HCMV pentameric complex of claim 17 , wherein said gL, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 7-9 and 31.

19. The purified HCMV pentameric complex of claim 17 , wherein said pUL128, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 13-15 and 33.

20. The purified HCMV pentameric complex of claim 17 , wherein said pUL130, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 16, 17, and 34.

21. The purified HCMV pentameric complex of claim 17 , wherein said pUL131A, or pentamer-forming fragment thereof, comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 18-20, and 35.

22. The purified HCMV pentameric complex of claim 17 , wherein said pentamer-forming fragment of HCMV gH comprises a sequence that is at least 80% identical to any one of SEQ ID NOs: 4, 6, 29, and 30.

23. The purified HCMV pentameric complex of claim 22 , wherein said pentamer-forming fragment of HCMV gH comprises a sequence selected from the group consisting of SEQ ID NOs: 4, 6, 29, and 30.

24. The purified HCMV pentameric complex of claim 23 , wherein said pentamer-forming fragment of HCMV gH comprises the sequence SEQ ID NO: 6.

25. The composition of claim 1 , wherein said gL, or pentamer-forming fragment thereof, comprises a sequence that is at least 85% identical to any one of SEQ ID NOs: 7-9 and 31.

26. The composition of claim 1 , wherein said gL, or pentamer-forming fragment thereof, comprises a sequence that is at least 90% identical to any one of SEQ ID NOs: 7-9 and 31.

27. The composition of claim 1 , wherein said gL, or pentamer-forming fragment thereof, comprises a sequence that is at least 95% identical to any one of SEQ ID NOs: 7-9 and 31.

28. The composition of claim 1 , wherein said pUL128, or pentamer-forming fragment thereof, comprises a sequence that is at least 85% identical to any one of SEQ ID NOs: 13-15 and 33.

29. The composition of claim 1 , wherein said pUL128, or pentamer-forming fragment thereof, comprises a sequence that is at least 90% identical to any one of SEQ ID NOs: 13-15 and 33.

30. The composition of claim 1 , wherein said pUL128, or pentamer-forming fragment thereof, comprises a sequence that is at least 95% identical to any one of SEQ ID NOs: 13-15 and 33.

31. The composition of claim 1 , wherein said pUL130, or pentamer-forming fragment thereof, comprises a sequence that is at least 85% identical to any one of SEQ ID NOs: 16, 17, and 34.

32. The composition of claim 1 , wherein said pUL130, or pentamer-forming fragment thereof, comprises a sequence that is at least 90% identical to any one of SEQ ID NOs: 16, 17, and 34.

33. The composition of claim 1 , wherein said pUL130, or pentamer-forming fragment thereof, comprises a sequence that is at least 95% identical to any one of SEQ ID NOs: 16, 17, and 34.

34. The composition of claim 1 , wherein said pUL131A, or pentamer-forming fragment thereof, comprises a sequence that is at least 85% identical to any one of SEQ ID NOs: 18-20, and

Assignees

• Glaxosmithkline Biologicals Sa
• Glaxosmithkline Biologicals Sa

Inventors

• Carfi Andrea
• Wen Yingxia

Classifications

• C07K14/005 Primary

  from viruses · CPC title

• C12N2710/16151

  Methods of production or purification of viral material · CPC title

• A61K2039/545

  characterised by the dose, timing or administration schedule · CPC title

• C12N2710/16134

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title

• A61K2039/572

  cytotoxic response · CPC title