Chromatographic purification of adeno-associated virus (AAV)
AssigneeGenzyme Corporation
InventorsCatherine E. O'Riordan, Amy E. Erickson, Alan E. Smith
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Abstract
The present invention relates to the purification of large scale quantities of active (infectious) adenovirus and AAV, especially for use in therapeutic applications. In particular, the invention provides improved methods for contacting such viruses with suitable chromatographic materials in a fashion such that any damage to the virus, particularly to surface components thereof, resulting from contact with such chromatographic materials is minimized or eliminated. The result is the ability to rapidly and efficiently purify commercial level quantities of active (infectious) virus suitable for use in therapeutic applications, e.g. gene transfer/therapy procedures.
Claims4
An improved method for purification of adeno-associated virus (AAV) from a composition containing both AAV and adenovirus, wherein the improvement comprises inclusion of a step for inactivation and removal of adenovirus, said step comprising mixing the composition with a chromatographic matrix material having pores which are approximately the size of adenovirus, such that adenovirus is inactivated, said chromatographic matrix material being selected from the group consisting of:
- a. "macroporous" resins, such as macroporous DEAE resins and the BioRad macroporous series (Biorad, Melville, N.Y.);
- b. heparinized polymers, such as Heparin Agarose (4% cross-linked) (Sigma Chemical, St. Louis, Mo.);
- c. virus-specific antibodies;
- such as anti-rep antibodies;
- d. sulfated resins, such as Sterogene-S (Sulfated Hi Flow) (Sterogene, Carlsbad, Calif.), Spherilose-S (Isco, Lincoln, Nebr.), and Cellufine.RTM. sulfate (Amicon, Beverly, Mass.);
- e. hydroxyapatite resins, such as ceramic hydroxyapatite resins from Biorad (Melville, N.Y.);
- f. a series of column separations including the use of ceramic hydroxyapatite, DEAE ion-exchange, Cellufine.RTM. sulfate, and, optionally, zinc chelate chromatography.
Dependencies are inferred from the claim text.