BATCHES OF RECOMBINANT ADENOVIRUS WITH ALTERED TERMINAL ENDS

• US 20140073032A1
• Filed: 03/11/2013
• Published: 03/13/2014
• Est. Priority Date: 03/12/2012
• Status: Active Grant

• Current Assignee: Crucell Holland B.V (Johnson & Johnson)
• Original Assignee: Crucell Holland B.V (Johnson & Johnson)
• Inventors: Custers, Jerome H. H. V., Vellinga, Jort
• Granted Patent: US 8,932,607 B2
• Time in Patent Office: Days
• US Class Current: 435/239
• CPC Class Codes: C12N 2710/10321 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2710/10343 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2710/10351 Methods of production or purification of viral material
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector
• CPC Class Codes: C12N 2760/16121 Viruses as such, e.g. new isolates, mutants or their genomic sequences
• CPC Class Codes: C12N 2760/16151 Methods of production or purification of viral material
• CPC Class Codes: C12N 2760/16234 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

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

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 (nucleotides 1-8 of SEQ ID NO;

7).

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, wherein the adenovirus comprises a transgene.

7. The composition of claim 1, comprising at least 1×

10⁷ recombinant adenovirus particles.

18. The composition of claim 7, comprising at least 1×

10⁸ recombinant adenovirus particles.

19. The composition of claim 18, comprising at least 1×

10⁹ recombinant adenovirus particles.

20. The composition of claim 19, comprising at least 1×

10¹⁰ recombinant adenovirus particles.

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 molecular cloning step to exchange naturally occurring 5′

termini of an adenovirus genome with altered 5′

termini comprising as terminal nucleotides the polynucleotide;

CTATCTAT (nucleotides 1-8 of SEQ ID NO;

7),

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 (nucleotides 1-8 of SEQ ID NO;

7).

10. The method according to claim 8, wherein the batch comprises at least 1×

10⁷ recombinant adenovirus particles.

11. The method according to claim 8, wherein the recombinant adenovirus is a recombinant human adenovirus of serotype 5, 26, 35, 49, or 50.

12. The method according to claim 8, wherein the recombinant adenovirus is a recombinant human adenovirus of serotype 26 or 35.

13. The method according to claim 8, wherein the recombinant adenovirus lacks at least a portion of the E1 region.

14. The method according to claim 8, wherein the recombinant adenovirus comprises a transgene.

15. The method according to claim 8, further comprising purifying the recombinant adenovirus.

16. The method according to claim 15, further comprising formulating the recombinant adenovirus into a pharmaceutical composition.

17. The method according to claim 8, wherein step b) is performed in a bioreactor.

9. 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 (nucleotides 1-8 of SEQ ID NO;

7),

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 (nucleotides 1-8 of SEQ ID NO;

7).