Purification of herpes virus
The present disclosure provides a method to prepare purified enveloped viral particle preparations employing ion exchange chromatography and tangential flow filtration.
1. A composition comprising Herpes Simplex Virus (HSV) particles in a liquid stabilization buffer, wherein the liquid stabilization buffer comprises glutamate, 1-20 mM histidine and optionally glutamine, a salt, and a sugar, and wherein other than histidine and optionally glutamine, the liquid stabilization buffer does not contain any other amino acids.
2. The composition of claim 1 , wherein the liquid stabilization buffer comprises 20-75 mM potassium glutamate, 1-20 mM histidine, 50-250 mM salt, and 5-20% sugar.
3. The composition of claim 2 wherein the liquid stabilization buffer comprises 50 mM potassium glutamate, 10 mM histidine, 160 mM salt, and 10% sugar.
4. The composition of claim 3 , wherein the sugar is sucrose.
5. The composition of claim 2 , wherein the pH of the liquid stabilization buffer is about 7.5.
6. The composition of claim 2 , wherein the HSV is a replication defective HSV.
7. The composition of claim 6 , wherein the replication defective HSV is HSV529.
8. The composition of claim 2 , wherein the residual host cell DNA in said composition is less than 10 ng host cell DNA per 1×10 7 PFU.
9. The composition of claim 2 , wherein the composition contains greater than 1×10 7 PFU/mL.
10. The composition of claim 2 , wherein the composition contains between about 1×10 7 to 2×10 7 PFU/mL.
11. The composition of claim 2 , wherein the sugar is sucrose.
12. The composition of claim 3 , wherein the pH of the liquid stabilization buffer is about 7.5.
13. The composition of claim 12 , wherein the residual host cell DNA in said composition is less than 10 ng host cell DNA per 1×10 7 PFU.
14. The composition of claim 13 , wherein the composition contains greater than 1×10 7 PFU/mL.
15. The composition of claim 8 , wherein the composition is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.
16. The composition of claim 13 , wherein the composition is a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.
17. The composition of claim 1 , wherein the liquid stabilization buffer comprises potassium glutamate.
18. The composition of claim 1 , wherein the HSV is herpes simplex virus type-2 (HSV-2) or herpes simplex virus type-1 (HSV-1).