Virus preparations and methods

• US 6,267,967 B1
• Filed: 03/05/1999
• Issued: 07/31/2001
• Est. Priority Date: 03/05/1998
• Status: Expired due to Fees

• Current Assignee: Cantab Pharmaceuticals Research Limited
• Original Assignee: Cantab Pharmaceuticals Research Limited
• Inventors: Slater, Nigel Kenneth Harry, Johnston, Michael Denis, OʼKeeffe, Roderic Simon
• Primary Examiner(s): MOSHER, MARY
• Application Number:
  US09/263,140
• Time in Patent Office: 879 Days
• Field of Search: 435/239, 424/229.1, 424/231.1
• US Class Current: 424/229.1
• CPC Class Codes: A61K 39/12 Viral antigens
• CPC Class Codes: A61K 39/245 Herpetoviridae, e.g. herpes simplex virus
• CPC Class Codes: A61P 31/22 for herpes viruses
• CPC Class Codes: C12N 2710/16634 Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
• CPC Class Codes: C12N 2710/16651 Methods of production or purification of viral material
• 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

Herpesvirus preparations, e.g. cultured HSV type 2, e.g. genetically disabled virus for vaccine use, can be purified, e.g. for subsequent pharmaceutical formulation, with solid phase affinity reagents containing sulfate- or sulfonate-comprising binding groups, e.g. sulfated polysacharide groups, e.g. heparin or dextran sulfate, and eluting e.g. with salt solutions. The process can be combined with other culture and harvesting steps.

Claims

1. A process for purifying a herpesvirus preparation, which comprises (a) culturing host cells infected with the virus, (b) contacting the cells with an eluant without substantial cell breakage, and eluting the virus from said cells in said eluant, (c) contacting the herpesvirus-containing eluate to be purified, with an affinity binding reagent that comprises a solid phase carrying a sulfate or sulfonate-comprising binding group that can bind materials with affinity for heparin, thereby to bind said herpesvirus to the affinity binding reagent, (d) washing said affinity binding reagent carrying said herpesvirus, and (e) eluting said herpesvirus from said binding reagent.

2. A process according to claim 1, comprising the further step of afterwards formulating said herpesvirus with a pharmaceutically acceptable carrier or excipient, and optionally sterilising and freezing or freeze-drying the preparation.

3. A process according to claim 2 wherein the herpesvirus comprises infectious herpesvirus, and the resulting pharmaceutical formation is for use as a vaccine.

4. A process according to claim 2, in which (i) the herpesvirus is applied to the affinity reagent from a liquid containing sodium chloride or other pharmaceutically acceptable salt in a concentration greater than about 0.4 M, or containing sulfated or sulfonated polysaccharides, and (ii) the elution from the affinity reagent is carried out with a saline eluant or a sulfated or sulfonated polysaccharide eluant.

5. A process according to claim 1, wherein said affinity binding reagent carries binding groups containing sulfate and nonionic polar groups.

6. A process according to claim 5, wherein said affinity binding reagent carries heparin or dextran sulfate.

7. A process according to claim 1, wherein said affinity binding reagent carries sulfated polysaccharide groups.

8. A process according to claim 1, wherein said affinity binding reagent carries pendent polyarylamide chains substituted by sulfoisobutyl groups.

9. A process according to claim 1, wherein the herpesvirus preparation is infectious human herpes simplex virus type 2 (HSV-2).

10. A process according to claim 9, wherein the eluant used to eluate virus from the cells comprises sulfated polysaccharide, and wherein the virus is eluted from the affinity binding reagent with a high salt solution.

11. A process according to claim 1, wherein the eluant used to elute virus from the cells comprises a hypertonic salt solution, or comprises sulfated or sulfonated polysaccharide.

12. A process according to claim 1, further comprising separating the cells from their culture medium, after culturing host cells infected with the virus, and prior to contacting the cells with the eluant.

13. A process for purifying a herpesvirus preparation, comprising:

(a) contacting a herpesvirus-containing composition with an affinity binding reagent, thereby to bind the herpesvirus to the affinity binding reagent, wherein the herpesvirus-containing composition is a liquid which comprises sodium chloride or other pharmaceutically acceptable salt in a concentration greater than about 0.4 M, or a liquid which comprises a sulfated or sulfonated polysaccharide, and the affinity binding reagent comprises a solid phase carrying a sulfate- or sulfonate-comprising binding group that can bind materials with affinity for heparin;

(b) washing said affinity binding reagent carrying said herpesvirus; and

(c) eluting said herpesvirus from said binding reagent.

14. A process according to claim 13, comprising the further step of afterwards formulating said herpesvirus with a pharmaceutically acceptable carrier or excipient, and optionally sterilizing and freezing or freeze-drying the preparation.

15. A process according to claim 14, wherien the herpesvirus comprises infectious herpesvirus, and the resulting pharmaceutical formulation is for use as a vaccine.

16. A process according to claim 13, wherein said affinity binding reagent carries binding groups containing sulfate and nonionic polar groups.

17. A process according to claim 13, wherein said affinity binding reagent carries sulfated polysaccharide groups.

18. A process according to claim 13, wherein said affinity binding reagent carries pendent polyacrylemide chains substituted by sulfoisobutyl groups.

19. A process according to claim 13, wherein the herpesvirus preparation is infectious human herpes simplex virus type 2 (HSV-2).