ICP0-mediated enhanced expression system

Patent metadata

Field	Value
Publication number	US-11286465-B2
Application number	US-201816230990-A
Country	US
Kind code	B2
Filing date	Dec 21, 2018
Priority date	May 13, 2015
Publication date	Mar 29, 2022
Grant date	Mar 29, 2022

Abstract

Methods and compositions for increasing the production of recombinant proteins by introducing ICP0 to cells capable of producing a recombinant protein are encompassed. In one method, the recombinant protein is a protein that is required for the replication of a replication defective virus, wherein the recombinant protein is provided to the replication defective virus in trans.

First claim

What is claimed is:

1. A method of increasing heterologous recombinant protein expression from a viral promoter in vitro, comprising:
introducing by transfection a herpes simplex virus (HSV) infected cell polypeptide zero (ICP0) to a cell comprising a transgene encoding the heterologous protein operably linked to the viral promoter, wherein the cell is a mammalian cell and is capable of expressing the recombinant protein from the viral promoter; and
isolating the recombinant protein from the cell, wherein:
(i) the recombinant protein is non-viral; and/or
(ii) the ICP0 is introduced by transfection with a plasmid comprising a sequence encoding ICP0 operably linked to a CMV promoter, an SV40 promoter, or a non-inducible promoter comprising a TATA box, GC-box, CCAAT box, B recognition element, and an initiator element; the heterologous recombinant protein is heterologous relative to the cell; and the heterologous protein is expressed by 24 hours after introduction of ICP0.

2. The method of claim 1 , wherein the recombinant protein is an antibody, a vaccine antigen, a hormone, or an enzyme.

3. The method of claim 1 , wherein the recombinant protein is a mammalian protein.

4. The method of claim 1 , wherein the recombinant protein is non-viral.

5. The method of claim 1 , wherein the ICP0 is introduced by transfection with a plasmid encoding ICP0 and comprising a non-inducible promoter, and the non-inducible promoter is a CMV promoter, an SV40 promoter, or a promoter comprising a TATA box, GC-box, CCAAT box, B recognition element, and an initiator element.

6. The method of claim 1 , wherein the cell is a Vero, BHK, CHO, HKB, HEK, NSO and U-2 OS, WI-38, MRC-5, MDCK, FRhL-2, or PERC6 cell.

7. The method of claim 1 , wherein the cell is a Vero cell.

Assignees

• Sanofi Pasteur Biologies Llc
• Sanofi Pasteur Inc

Inventors

• Delagrave Simon
• Lai Charchang

Classifications

• C12N2710/16651

  Methods of production or purification of viral material · CPC title

• C12N2710/16634

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

• C12N2710/16622

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title

• C12N7/00 Primary

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• A61K39/12

  Viral antigens · CPC title