Claims
- 1. 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.
- 2. The method of claim 1, wherein the chromatographic matrix material comprises a macroporous DEAE resin.
- 3. The method of claim 1, wherein the chromatographic matrix material comprises a macroporous hydroxyapatite.
- 4. The method of claim 1, wherein the chromatographic matrix material comprises a cellulose or silica membrane resin in conjunction with a macroporous resin.
Parent Case Info
The present application is a national stage application filed under 35 USC chapter 371, from the international application No. PCT/US96/13872, filed on Aug. 30, 1996, which was a continuation in part of U.S. provisional patent application Ser. No. 60/002,967, filed on Aug. 30, 1995, now abandoned.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
102e Date |
371c Date |
PCT/US96/13872 |
8/30/1996 |
|
|
6/29/1998 |
6/29/1998 |
Publishing Document |
Publishing Date |
Country |
Kind |
WO97/08298 |
3/6/1997 |
|
|
US Referenced Citations (1)
Number |
Name |
Date |
Kind |
5837520 |
Shabram et al. |
Nov 1998 |
|
Foreign Referenced Citations (4)
Number |
Date |
Country |
0187894 |
Jul 1986 |
EPX |
1168015 |
Apr 1964 |
DEX |
1951800 |
Apr 1971 |
DEX |
1364342 |
Jan 1988 |
RUX |
Non-Patent Literature Citations (7)
Entry |
Tamayose et al. Human Gene Therapy. 1996, vol. 7, pp. 507-513, Mar. 1, 1996. |
Flotte et al., 1995, Gene Therapy 2:29-37. |
Chiorini et al., 1995, Human Gene Therapy 6:1531-1541. |
Fisher et al., 1996, J. Virol. 70:520-532. |
Hughe et al., 1995, Human Gene Therapy 6:1403-1416. |
Ivanov, A.E., et al., 1990, Bioorg. Khim. 16:1028-1039. |
O'Neil et al., 1993. Bio/technology 11:173-178. |