Virus purification
Disclosed herein are methods and exemplary compositions associated with virus purification, exemplary aspects of which may include harvesting viral and antigenic substances from source organisms; and a purification platform comprising chemical separation and size-difference separation for the removal of contaminants, debris and impurities from the viral and protein (e.g. antigenic, including influenza hemagglutinin antigens) substances, as well as their concentration and collection.
1. A method for purifying at least one virus harvested from a source organism, comprising:
removing cellular debris from the virus by passing the at least one virus through a tangential flow filtration apparatus comprising a porous ceramic filter;
concentrating the virus; and
performing separations comprising sub-steps of separating host cell contaminants from the virus, subjecting the virus to affinity chromatography to elute the virus, and subjecting the virus to multi-modal chromatography to separate residual impurities from the virus based on size differences between the impurities as retentate and the virus as permeate.
2. The method of claim 1 , further comprising harvesting the virus from a source organism, wherein the source organism is a plant.
3. The method of claim 1 , further comprising harvesting the virus from a source organism, wherein the source organism is selected from the group consisting of bacterial, algal, yeast, insect, and mammalian organisms.
4. The method of claim 1 , wherein concentrating the at least one virus comprises passing the at least one virus through a membranous filtration device comprising a porous membrane.
5. The method of claim 1 , wherein separating host cell contaminants from the virus comprises subjecting the virus to ion-exchange chromatography.
6. The method of claim 5 , further comprising collecting the virus after the ion-exchange chromatography sub-step or the multi-modal chromatography sub-step.
7. The method of claim 1 , further comprising diluting the virus to a concentration of less than about 50 micrograms per milliliter and inactivating the virus by exposing it to light in the UV spectrum.
8. The method of claim 7 , wherein the light has an energy density between about 2400 J/m 2 and 5142 J/m 2 .
9. The method of claim 1 , wherein the source organism is a plant.
10. The method of claim 1 , wherein the method further comprises establishing a transmembrane pressure across the filter between about 10-20 psi.