Large-scale recombinant adeno-associated virus (rAAV) production and purification
US6593123B1 · kind B1 · utility
Assignee
Inventors
Key dates
| Filing date | Aug 7, 2000 |
| Grant date | Jul 15, 2003 |
| Priority date | — |
| Expiry date | Aug 7, 2020 |
Classification
- Technology area (CPC C)Chemistry; Metallurgy
- CPC primaryC12N2750/14151
- WIPO fieldBiotechnology
- WIPO sectorChemistry
Abstract
Methods are provided for large-scale purification of recombinant AAV (rAAV) virions that were produced in the absence of infectious adenovirus. Preferably, the rAAV is produced in a host cell line via triple-transfection with an accessory function vector, an AAV vector, and an AAV helper vector. The methods include preparing a lysate from the host cell line and passing that lysate over various combinations of ion exchange chromatography media and/or affinity chromatography media. The affinity chromatography medium is an AAV receptor or an antibody with binding affinity for AAV, e.g., heparin sulfate. A variety of cation exchange and anion exchange media are contemplated by the present invention. In certain embodiments, optional purification steps may be included, such as filtering the lysate through one or more filters, or treating the lysate with a nuclease.
Source: USPTO / EPO open patent data. Objective bibliographic and citation counts.