Claims
- 1. A process for purifying recombinant papillomavirus virus-like particles (VLPs) comprising:(a) contacting a partially purified VLP-containing cell lysate with a hydroxyapatite medium in a chromatography column, under conditions such that the VLPs bind to the hydroxyapatite medium; (b) eluting the bound VLPs with a solution comprising phosphate anions; and (c) recovering the eluted VLPs.
- 2. A process according to claim 1 wherein the VLPs consist essentially of L1 protein.
- 3. A process according to claim 1 wherein the VLPs are human papillomavirus (HPV) VLPs.
- 4. A process according to claim 3 wherein the VLPs are selected from the group consisting of: HPV type 6a, HPV type 6b, HPV type 11, HPV type 16, HPV type 18, HPV type 31, HPV type 33, and HPV type 45.
- 5. A process according to claim 4 wherein the eluted VLPs are at least 75% pure.
- 6. A process according to claim 4 wherein the eluted VLPs are at least 90% pure.
- 7. A process according to claim 1 wherein the VLPs comprise L1 protein and a L2:fusion protein.
- 8. A process according to claim 7 wherein the L2:fusion protein has a L2 portion which is less than full length.
- 9. A process of making a purified human papillomavirus VLP product, suitable for use in a human vaccine comprising:(a) partially purifying a cell lysate, wherein the cell lysate is from yeast cells which have been transformed to express HPV L1 VLPs; (b) contacting the partially purified VLP-containing cell lysate with a hydroxyapatite medium in a chromatography column, under conditions such that the VLPs bind to the hydroxyapatite medium; (c) eluting the bound VLPs with a solution comprising phosphate anions; and (d) recovering the eluted VLPs.
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a U.S. national application of PCT/US99/17930, filed Aug. 10, 1999 and which published as WO 00/09671, Feb. 24, 2000, and which claims the benefit of U.S. Provisional Patent Application No. 60/096,568, filed Aug. 14, 1998.
PCT Information
Filing Document |
Filing Date |
Country |
Kind |
PCT/US99/17930 |
|
WO |
00 |
Publishing Document |
Publishing Date |
Country |
Kind |
WO00/09671 |
2/24/2000 |
WO |
A |
US Referenced Citations (2)
Number |
Name |
Date |
Kind |
4612283 |
Sugahara et al. |
Sep 1986 |
A |
5085781 |
Tsuru et al. |
Feb 1992 |
A |
Foreign Referenced Citations (6)
Number |
Date |
Country |
WO 9531532 |
Nov 1995 |
WO |
9609375 |
Mar 1996 |
WO |
9615247 |
May 1996 |
WO |
9629413 |
Sep 1996 |
WO |
9630520 |
Oct 1996 |
WO |
WO 9708298 |
Mar 1997 |
WO |
Non-Patent Literature Citations (2)
Entry |
Scopes, Robert K. Protein Purification, Principles and Practise, Second Edition. Springer-Verlag, New York, 1987. pp. 173-176.* |
Tsuru, S. et al. “Adsorption and Preparation of Human Viruses Using Hydroxyapatite Column”, Bio-Medical Materials and Engineering, 1991, vol. 1, pp. 143-147. |
Provisional Applications (1)
|
Number |
Date |
Country |
|
60/096568 |
Aug 1998 |
US |