ICP0-mediated enhanced expression system
AssigneeSanofi Pasteur Biologics, LLC
InventorsSimon Delagrave, CharChang Lai
Life of this patent
Estimating the term…
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.
Claims16
A method for producing a replication defective Herpes simplex virus (HSV) in cell culture comprising:
- a) providing a cell comprising a nucleic acid encoding at least one recombinant protein, wherein the recombinant protein is required for replication of said replication defective HSV;
- b) introducing a herpes simplex virus infected cell polypeptide zero (ICP0) to the cell by transduction with a virus other than HSV or transfection, thereby increasing the expression of the recombinant protein;
- c) introducing a replication defective HSV to the cell, wherein the recombinant protein complements the replication defective virus in trans;
- d) replicating said replication defective HSV in the cell;
- and
- e) isolating the replication defective HSV from the cell;
- wherein the recombinant protein is ICP8 and/or pUL5.
A method for producing replication-defective Herpes simplex virus 2 (HSV-2) vaccine HSV529 comprising the following steps, in order:
- a) introducing ICP0 to AV529-19 cells, wherein ICP0 increases the expression of at least one of ICP8 or pUL5 in the AV529-19 cells;
- b) infecting the AV529-19 cells with HSV529;
- c) replicating HSV529 in the cell;
- and
- d) isolating HSV529 from the cell.
A method for increasing the yield of replication defective HSV-2 vaccine HSV529 comprising the following steps, in order:
- a) introducing ICP0 to AV529-19 cells, wherein ICP0 increases the expression of at least one of ICP8 or pUL5 in the AV529-19 cells;
- b) infecting the AV529-19 cells with HSV529;
- c) replicating HSV529 in the cell;
- and
- d) isolating HSV529 from the cell.
Dependencies are inferred from the claim text.