Aseptic purification process for viruses
Described herein are processes for the purification of viruses and compositions thereof.
1. A process of purification of virus particles comprising the steps of:
(a) providing a liquid medium comprising virus particles, wherein the virus particles are greater than about 100 nm in diameter;
(b) contacting the virus particles with a solid-phase matrix comprising a ligand-activated core and an inactive shell comprising pores having a molecular weight cut off that excludes the virus particles from entering the ligand-activated core, and wherein a molecule smaller than the molecular weight cut off of the pores can enter the ligand-activated core; and
(c) separating the solid-phase matrix from the virus particles by filtration to produce a final virus preparation;
wherein the solid-phase matrix is added to the liquid medium comprising the virus particles as a slurry at a final concentration between 2.5% (v/v) and 30% (v/v); and
wherein each step of the process is performed aseptically.
2. The process of claim 1 , wherein the liquid medium comprising the virus particles is subjected to one or more pre-purification step(s) prior to step (b).
3. The process of claim 1 , wherein the virus particles are about 200 nm, 300 nm, 400 nm, 500 nm or more in diameter.
4. The process of claim 1 , wherein the molecule entering the core of the solid-phase matrix has a molecular weight less than 700 kDa.
5. The process of claim 1 , wherein the ligand of the ligand-activated core of the solid-phase matrix is octylamine.
6. The process of claim 1 , wherein the relative reduction of impurity of the final virus preparation relative to the liquid medium comprising the plurality of the viruses or virus particles is in a range from 60 to 95%.
7. The process of claim 2 , wherein the pre-purification step comprises
(i) digesting host cell genomic DNA in the liquid medium comprising the plurality of the viruses or virus particles by enzymatic treatment; and/or
(ii) ultra/diafiltration of the liquid medium comprising the plurality of the viruses or virus particles using a hollow fiber membrane having a pore size equal or greater than 750 kDa.
8. The process of claim 1 , wherein the process is followed by one or more aseptic filtration step(s).
9. The process of claim 1 , wherein the residual impurity of the final virus preparation is less than 1%.
10. The process of claim 1 , wherein the virus is propagated in a cell line selected from the group consisting of an EB66 cell line, a Vero cell line, a Vero-αHis cell line, a HeLa cell line, a HeLa-S3 cell line, a 293 cell line, a PC12 cell line, a CHO cell line, a 3T3 cell line, a PerC6 cell line, an MDSK cell line, a chicken embryonic fibroblast cell line, a duck cell line, and a diploid avian cell line.
11. The process of claim 10 , wherein said cell line is the EB66 cell line.
12. The process of claim 1 , wherein the virus belongs to a virus family selected from the group consisting of Paramyxoviridae, Orthomyxoviridae, Flaviviridae, Filoviridae, Arenaviridae, Rhabdoviridae, and Coronaviridae.
13. The process of claim 12 , wherein the virus belongs to the Paramyxoviridae virus family.
14. The process of claim 13 , wherein the virus is a measles virus.
15. A method for manufacturing a composition for immunization against a viral infection comprising purifying virus particles according to the process of claim 1 .
16. A process of purification of virus particles, wherein the virus belongs to the Paramyxoviridae family, comprising the steps of:
(a) providing a liquid medium comprising Paramyxoviridae family virus particles, wherein the Paramyxoviridae family virus particles are greater than about 100 nm in diameter;
(b) contacting the Paramyxoviridae family virus particles with a solid-phase matrix comprising a ligand-activated core and an inactive shell comprising pores having a molecular weight cut off that excludes the Paramyxoviridae family virus particles from entering the ligand-activated core, and wherein a molecule smaller than the molecular weight cut off of the pores can enter the ligand-activated core; and
(c) separating the solid-phase matrix from the Paramyxoviridae family virus particles by filtration to produce a final Paramyxoviridae family virus preparation;
wherein the solid-phase matrix is added to the liquid medium comprising the Paramyxoviridae family virus particles as a slurry at a final concentration between 2.5% (v/v) and 30% (v/v), and
wherein each step of the process is performed aseptically.
17. A process of purification of virus particles of a measles virus comprising the steps of:
(a) providing a liquid medium comprising measles virus particles, wherein the measles virus particles are greater than about 100 nm in diameter;
(b) contacting the measles virus particles with a solid-phase matrix comprising a ligand-activated core and an inactive shell comprising pores having a molecular weight cut off that excludes the measles virus particles from entering the ligand-activated core, and wherein a molecule smaller than the molecular weight cut off of the pores can enter the ligand-activated core; and
(c) separating the solid-phase matrix from the measles virus particles by filtration to produce a final measles virus preparation;
wherein the solid-phase matrix is added to the liquid medium comprising the measles virus particles as a slurry at a final concentration between 2.5% (v/v) and 30% (v/v), and
wherein each step of the process is performed aseptically.