Adenovirus derived helper virus for enhancing recombinant parvovirus production

Patent metadata

Field	Value
Publication number	US-9803219-B2
Application number	US-201213627554-A
Country	US
Kind code	B2
Filing date	Sep 26, 2012
Priority date	Mar 26, 2010
Publication date	Oct 31, 2017
Grant date	Oct 31, 2017

Abstract

Described is an adenovirus derived helper virus which may comprise the adenoviral DNA sequences for E2a, S4 (orf6), the VA1 RNA gene, and the parvovirus VP capsid gene unit. Also described is a method of efficiently preparing a recombinant parvovirus (particle) which is based on the use of various adenoviral derived helper viruses/vectors.

First claim

What is claimed is:

1. An isolated NB324K (NBK) cell comprising:
(a) an adenovirus derived helper virus comprising:
(i) adenoviral DNA sequences:
(a) an adenoviral E2a gene;
(b) an adenoviral E4(orf6) gene; and
(c) an adenoviral VAI RNA gene; and

(ii) a recombinant parvovirus VP capsid gene unit of H-1 or MVM; and

(b) a recombinant parvovirus derived from H-1 or MVM that lacks the VP1 and VP2 genes,
wherein the NBK cell produces at least two fold higher an amount of recombinant parvovirus as compared to an NBK cell which does not comprise the adenovirus derived helper virus of (a).

2. The cell of claim 1 , wherein the expression of the parvovirus VP capsid gene unit is under the control of a CMV promoter.

3. The cell of claim 1 , wherein the adenoviral DNA sequences are derived from Ad5.

4. The cell of claim 1 , wherein the adenovirus derived helper virus and the recombinant parvovirus contain a transgene.

5. The cell of claim 1 , wherein in the recombinant parvovirus the VP1 and VP2 genes are replaced by a transgene.

6. The cell of claim 5 , wherein expression of the transgene is under the control of the P38 promoter.

7. The cell of claim 5 , wherein the transgene is a gene encoding a marker protein.

8. The cell of claim 5 , wherein the transgene is a gene encoding a therapeutic or immunogenic polypeptide.

9. The cell of claim 8 , wherein the therapeutic protein is a cytotoxic polypeptide, cytokine and/or chemokine.

Assignees

• Deutsches Krebsforsch

Inventors

• El-Andaloussi Nazim
• Marchini Antonio
• Rommelaere Jean
• Leuchs Barbara
• Endele Max

Classifications

• C12N2750/14344

  Chimeric viral vector comprising heterologous viral elements for production of another viral vector · CPC title

• A61K35/13

  Tumour cells, irrespective of tissue of origin (tumour vaccines A61K39/00) · CPC title

• C12N2750/14351

  Methods of production or purification of viral material · CPC title

• C12N2710/10344

  Chimeric viral vector comprising heterologous viral elements for production of another viral vector · CPC title

• A61P35/00

  Antineoplastic agents · CPC title