Virus preparations and methods
View Patent ↗Methods are disclosed for the preparation of herpesvirus, such as herpes simplex virus type 2 for vaccine use. Such viruses can be grown on serum free or serum containing media and can be prepared from the virus containing culture supernatant or virus containing cells. The virus is prepared for subsequent pharmaceutical formulation by methods which may include treatment with solid phase affinity reagents containing sulfate- or sulfonate-comprising binding groups. Such sulfated polysaccharide groups as heparin or dextran sulfate may be used, and eluted with salt solutions. The process can be combined with other culture, harvesting and formulation steps.
1. A process for preparing a recombinant herpesvirus for a vaccine or therapy, which comprises:
(a) culturing host cells, selected from the group of host cells consisting of Vero, or MRC5 cells, infected with the virus,
(b) separating the medium from the infected cells and
(c) using the separated virus containing medium for further purification, immunotherapy or vaccination of an animal.
2. The process of claim 1 in which said host cells are selected from the group consisting of Vero and MRC5 cells.
3. The process of claim 1 in which said host cells are Vera cells.
4. The process of claim 1 wherein said herpesvirus is an HSV-2 which has the PK domain deleted.
5. The process of claim 1 wherein said herpesvirus is ICP10deltaPK.
6. The process of claim 1 wherein said culture is carried out in a serum free medium.
7. The process of claim 1 wherein said culture is carried out in a medium containing about 5 μg/mL protein or less.
8. The process of claim 1 where said culture is carried out in a medium with added epidermal growth factor.
9. The process of claim 1 where said culture is carried out in a medium with added insulin.
10. A process according to claim 1 , further comprising the step:
(d) formulating said herpesvirus with a pharmaceutically acceptable carrier, excipient or stabilizing agent.
11. A process according to claim 1 further comprising the step:
(e) sterilizing and freezing or freeze-drying the preparation.
12. A process according to claim 1 wherein the herpesvirus comprises infectious herpesvirus.
13. A process according to claim 1 wherein the resulting pharmaceutical formulation is for use as an immunotherapy.
14. A process for growing recombinant herpesvirus comprising the step:
(a) culturing host cells, selected from the group of cells consisting of Vero, or MRC5 cells, infected with the virus in a serum free medium.
15. The process of claim 14 in which said host cells are selected from the group consisting of Vero and MRC5 cells.
16. The process of claim 14 in which said host cells are Vero cells.