IP Library › Granted Patent US 8,932,607
Granted Patent B2
US 8,932,607 · App. 13/794,318 · Granted Jan 13, 2015

Batches of recombinant adenovirus with altered terminal ends

Inventors: Jerome H. H. V. Custers (Alphen aan den Rijn, NL); Jort Vellinga (Leiden, NL)
Assignee: Crucell Holland B.V.
C12N7/00C12N2760/16234C12N2760/16121C12N2750/14143C12N2760/16151C12N2710/10321C12N2710/10343
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Quick Facts
Patent No.
US 8,932,607
App. No.
13/794,318
Granted
Jan 13, 2015
Kind
B2
Abstract

Described is a composition comprising a plurality of recombinant adenovirus particles, being a recombinant human adenovirus of serotype 5, 26, 34, 35, 48, 49 or 50, or a recombinant simian adenovirus, characterized in that the genomes of essentially all adenovirus particles in the composition comprise as the 5′ terminal nucleotides the nucleotide sequence: CTATCTAT (nucleotides 1-8 of SEQ ID NO:7). Also described are methods to produce such compositions.

Claims (52)

1. A composition comprising adenovirus particles, wherein the adenovirus is a recombinant human adenovirus of serotype 5, 11a, 26, 34, 35, 48, 49 or 50, or a recombinant simian adenovirus, wherein the genomes of essentially all adenovirus particles in the composition have as the 5′ terminal nucleotides the polynucleotide: CTATCTAT, and wherein the adenovirus comprises a transgene.

2. The composition of claim 1 , wherein the adenovirus is a human adenovirus of serotype 5, 26, 35, 49, or 50.

3. The composition of claim 2 , wherein the adenovirus is a human adenovirus of serotype 26 or 35.

4. The composition of claim 1 , which is a pharmaceutical composition.

5. The composition of claim 1 , wherein the adenovirus lacks at least a portion of the E1 region.

6. The composition of claim 1 , comprising at least 1×10 7 recombinant adenovirus particles.

7. A method for preparing a batch of recombinant adenovirus particles that have essentially all identical polynucleotides in the 5′ termini of their genomes, the method comprising:

a) performing a molecular cloning step to exchange naturally occurring 5′ termini of an adenovirus genome with altered 5′ termini comprising as terminal nucleotides the polynucleotide: CTATCTAT,

b) propagating in host cells the recombinant adenovirus having the altered 5′ termini, and

c) harvesting the recombinant adenovirus to obtain a batch of recombinant adenovirus particles that essentially all have as the 5′ terminal nucleotides of their genomes the polynucleotide: CTATCTAT, and wherein the adenovirus comprises a transgene.

8. A method for preparing a batch of recombinant adenovirus particles that have essentially all identical polynucleotides in the 5′ termini of their genomes, the method comprising:

a) performing a plaque purification of an adenovirus, wherein the recombinant adenovirus is a recombinant human adenovirus of serotype 5, 11a, 26, 34, 35, 48, 49 or 50, or a recombinant simian adenovirus, to isolate an adenovirus or recombinant adenovirus from a single plaque, wherein the adenovirus or recombinant adenovirus has as the 5′ terminal nucleotides of its genome polynucleotide: CTATCTAT,

b) propagating in host cells a recombinant adenovirus obtained from the single plaque of step a), and

c) harvesting the recombinant adenovirus to obtain a batch of recombinant adenovirus particles that essentially all have as the 5′ terminal nucleotides of their genomes the polynucleotide: CTATCTAT, and wherein the adenovirus comprises a transgene.

9. The method according to claim 7 , wherein the batch comprises at least 1×10 7 recombinant adenovirus particles.

10. The method according to claim 7 , wherein the recombinant adenovirus is a recombinant human adenovirus of serotype 5, 26, 35, 49, or 50.

11. The method according to claim 7 , wherein the recombinant adenovirus is a recombinant human adenovirus of serotype 26 or 35.

12. The method according to claim 7 , wherein the recombinant adenovirus lacks at least a portion of the E1 region.

13. The method according to claim 7 , further comprising purifying the recombinant adenovirus.

14. The method according to claim 13 , further comprising formulating the recombinant adenovirus into a pharmaceutical composition.

15. The method according to claim 7 , wherein step b) is performed in a bioreactor.

16. The composition of claim 6 , comprising at least 1×10 8 recombinant adenovirus particles.

17. The composition of claim 16 , comprising at least 1×10 9 recombinant adenovirus particles.

18. The composition of claim 17 , comprising at least 1×10 10 recombinant adenovirus particles.

19. The composition of claim 1 , wherein the adenovirus is a human adenovirus of serotype 35.

20. The composition of claim 1 , wherein the adenovirus is a human adenovirus of serotype 26.

21. The composition of claim 2 , which is a pharmaceutical composition.

22. The composition of claim 3 , which is a pharmaceutical composition.

23. The composition of claim 19 , which is a pharmaceutical composition.

24. The composition of claim 20 , which is a pharmaceutical composition.

25. The composition of claim 2 , wherein the adenovirus lacks at least a portion of the E1 region.

26. The composition of claim 3 , wherein the adenovirus lacks at least a portion of the E1 region.

27. The composition of claim 19 , wherein the adenovirus lacks at least a portion of the E1 region.

28. The composition of claim 20 , wherein the adenovirus lacks at least a portion of the E1 region.

29. The composition of claim 23 , wherein the adenovirus lacks at least a portion of the E1 region.

30. The composition of claim 24 , wherein the adenovirus lacks at least a portion of the E1 region.

31. The composition of claim 2 , comprising at least 1×10 10 recombinant adenovirus particles.

32. The composition of claim 3 , comprising at least 1×10 10 recombinant adenovirus particles.

33. The composition of claim 4 , comprising at least 1×10 10 recombinant adenovirus particles.

34. The composition of claim 5 , comprising at least 1×10 10 recombinant adenovirus particles.

35. The composition of claim 19 , comprising at least 1×10 10 recombinant adenovirus particles.

36. The composition of claim 20 , comprising at least 1×10 10 recombinant adenovirus particles.

37. The composition of claim 21 , comprising at least 1×10 10 recombinant adenovirus particles.

38. The composition of claim 22 , comprising at least 1×10 10 recombinant adenovirus particles.

39. The composition of claim 23 , comprising at least 1×10 10 recombinant adenovirus particles.

40. The composition of claim 24 , comprising at least 1×10 10 recombinant adenovirus particles.

41. The composition of claim 25 , comprising at least 1×10 10 recombinant adenovirus particles.

42. The composition of claim 26 , comprising at least 1×10 10 recombinant adenovirus particles.

43. The composition of claim 27 , comprising at least 1×10 10 recombinant adenovirus particles.

44. The composition of claim 28 , comprising at least 1×10 10 recombinant adenovirus particles.

45. The composition of claim 29 , comprising at least 1×10 10 recombinant adenovirus particles.

46. The composition of claim 30 , comprising at least 1×10 10 recombinant adenovirus particles.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 3, 2013
From: CUSTERS, JEROME H.H.V.; VELLINGA, JORT
To: CRUCELL HOLLAND B.V.
Reel/Frame 030532/0407 →
Continuity (2)
Provisional Application 61609678 · Mar 12, 2012
Related Publication 20140073032A1 · Mar 13, 2014