Production and applications for polyvalent vaccines against diseases caused by papilloma viruses

• US 20050026257A1
• Filed: 02/13/2004
• Published: 02/03/2005
• Est. Priority Date: 07/30/2001
• Status: Active Grant

• Current Assignee: Deutsches Krebsforschungszentrum Stiftung des Offentlichen Rechts
• Original Assignee: Deutsches Krebsforschungszentrum Stiftung des Offentlichen Rechts
• Inventors: Muller, Martin, Gissmann, Lutz, Pohlmeyer, Kai
• Granted Patent: US 7,354,714 B2
• Time in Patent Office: Days
• US Class Current: 435/70.210
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: C07K 16/084 Papovaviridae, e.g. papillomavirus, polyomavirus, SV40, BK virus, JC virus
• CPC Class Codes: C12N 2710/20034 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Abstract

A vaccine against disease caused by papilloma viruses is described in certain embodiments, as well as certain vectors, obtainable by the following methods: (a) one or more expression vectors that contain the DNA code for a structural protein of papilloma viruses or a fragment thereof are injected into mammals, whereby in at least some of the expression vectors randomly generated heterologous sequences are inserted into the DNA code (b) serums are obtained from the mammals and these are tested for the presence of antibodies against particles of various papilloma virus types, and (c) using the serums tested, the structural protein gene clones are identified that code for a polyvalent vaccine, and (d) the vaccine is produced from them. Procedures for producing a vaccine is also described, together with its use for vaccination against diseases caused by papilloma viruses.

Claims

1. A method of making an antibody against a plurality of papilloma virus types, comprising:

harvesting antibodies from a mammal following introduction into the mammal of a plurality of DNA clones, wherein each DNA clone comprises an expression vector comprising DNA that encodes at least a portion of a structural protein of a papilloma virus.

2. The method of claim 1 further comprising testing the mammal to detect an antibody against a papilloma virus generated by introducing the DNA clone into the mammal.

3. The method of claim 1, wherein the papilloma virus is a human pathogenic papilloma virus.

4. The method of claim 1, wherein the structural protein is L1.

5. The method of claim 1 wherein the at least a portion of a structural protein of a papilloma virus comprises at least one randomly generated heterologous sequence.

6. The method of claim 5 wherein the DNA clone heterologous sequences have a length of between 6 and 200 bases.

7. The method of claim 5 wherein the structural protein is L1 protein, the DNA clone comprises DNA that encodes for a hyper-variable portion of the L1 protein, and wherein the at least one randomly generated heterologous sequence comprises a heterologous sequence inserted into the hyper-variable portion of the L1 protein.

8. The method of claim 5, wherein the least one randomly generated heterologous sequence is disposed in a surface loop of the structural protein.

9. The method of claim 1, wherein the number of DNA clones is between 10 and 10,000.

10. The method of claim 1, wherein the number of DNA clones is at least 25.

11. A library of DNA vectors, comprising:

a plurality of DNA clones each encoding and comprising a sequence that encodes at least a portion of a structural protein of a papilloma virus, wherein the sequence further comprises a randomly generated heterologous portion having a length of at least three nucleotides.

12. The library of claim 11, wherein the structural protein is L1.

13. The library of claim 11, wherein the randomly generated heterologous portion is disposed in a surface loop of the structural protein.

14. The library of claim 11, wherein the heterologous sequences have a length of between 3 and 200 bases.

15. The library of claim 11, wherein the number of expression vectors is between 10 and 10,000.

16. The library of claim 11, wherein the number of structural protein gene clones is at least 25.

17. The library of claim 16, wherein the structural protein is L1.

18. The library of claim 17, wherein the at least a portion of the L1 structural protein is a hyper-variable portion of L1 protein.

19. An animal, the animal comprising a plurality of DNA clones each encoding and comprising a sequence that encodes at least a portion of a structural protein of a papilloma virus.

20. The animal of claim 19, wherein the sequence further comprises a randomly generated heterologous portion having a length of at least three nucleotides.

21. The animal of claim 19, wherein the structural protein is L1.

22. The animal of claim 19, wherein the heterologous sequences have a length of between 3 and 200 bases.

23. The animal of claim 19, wherein the number of DNA clones is between 10 and 10,000.

24. The animal of claim 19, wherein the number of DNA clones is at least 25.

25. The animal of claim 19, wherein the randomly generated heterologous portion is disposed in a surface loop of the structural protein.

26. A method of creating an antibody, the method comprising introducing a plurality of DNA clones into an animal, with each DNA clone encoding and comprising a sequence that encodes at least a portion of a structural protein of a papilloma virus.

27. The method of claim 26, wherein the sequence further comprises a randomly generated heterologous portion having a length of at least three nucleotides.

28. The method of claim 26, wherein the structural protein is L1.

29. The method of claim 26, wherein the heterologous sequences have a length of between 3 and 200 bases.

30. The method of claim 26, wherein the number of DNA clones is between 10 and 10,000.

31. The method of claim 26, wherein the number of DNA clones is at least 25.

32. The method of claim 26, further comprising screening serum from the animal for the presence of at least one antibody that is reactive against a plurality of papilloma virus types.

33. The method of claim 32, wherein the screening method comprises a high-throughput method of detecting the antibodies.

34. The method of claim 32, wherein a plurality of animals are used, and each of the plurality of animals receives a plurality of DNA clones.

35. The method of claim 32, wherein the screening method comprises ELISA.

36. An antibody comprising a specific binding for at least two types of human papilloma virus.

37. The antibody of claim 36, wherein the at least two types of human papilloma virus include HPV 16 and HPV 18.

38. A method of making a library of nucleic acid vectors, comprising:

preparing a plurality of nucleic acid clones each encoding and comprising a nucleic acid sequence that encodes at least a portion of a structural protein of a papilloma virus, wherein the sequence further comprises a randomly generated heterologous portion.

39. The method of claim 38 wherein the nucleic acids comprise DNA.

40. The library of claim 38, wherein the structural protein is L1.

41. The library of claim 38, wherein the randomly generated heterologous portion is disposed in a surface loop of the structural protein.

42. The library of claim 38, wherein the heterologous sequences have a length of between 3 and 200 bases.

43. The library of claim 38, wherein the number of nucleic acid clones is between 10 and 10,000.

44. The library of claim 38, wherein the number of nucleic acid clones is at least 25.

45. The library of claim 38, wherein the structural protein is L1.

46. The library of claim 45, wherein the at least a portion of the L1 structural protein is a hyper-variable portion of L1 protein.