Title:
METHOD OF SEPARATING EXTRA-CHROMOSOMAL DNA FROM OTHER CELLULAR COMPONENTS
Document Type and Number:
WIPO Patent Application WO2002042317
Kind Code:
A3
Abstract:
The invention relates to a method of separating extra-chromosomal DNA from RNA. It also relates to DNA produced by the method and pharmaceuticals derived from such DNA, for example, DNA vaccines. The method comprises separating extra-chromosomal DNA from RNA without first digesting the RNA, and comprises: i) lysing cells comprising extra-chromosomal DNA to form a lysate; ii) clarifying the lysate; and iii) subjecting the clarified lysate to tangential flow ultra filtration under conditions, which allow a substantial amount of the RNA to pass through a membrane whilst the extra-chromosomal DNA is retained. The preferred conditions include the use of a diafiltration buffer of low conductivity and diafiltration volumes of over 10 volume equivalents.
Inventors:
CHARLTON HENRY (GB)
EON-DUVAL ALEX (GB)
MACDUFF ROBERT (GB)
EON-DUVAL ALEX (GB)
MACDUFF ROBERT (GB)
Application Number:
PCT/GB2001/005114
Publication Date:
February 06, 2003
Filing Date:
November 20, 2001
Export Citation:
Assignee:
GLAXO GROUP LTD (GB)
CHARLTON HENRY (GB)
EON-DUVAL ALEX (GB)
MACDUFF ROBERT (GB)
CHARLTON HENRY (GB)
EON-DUVAL ALEX (GB)
MACDUFF ROBERT (GB)
International Classes:
G01N30/02; A61K39/00; A61K48/00; B01D61/14; B01D71/16; B01D71/20; B01D71/26; B01D71/30; B01D71/44; B01D71/56; B01D71/68; B01J20/281; C12N15/09; C12N15/10; G01N30/26; G01N30/74; G01N30/88; (IPC1-7): C07H1/08; C12N15/10; A61K31/711
Domestic Patent References:
| WO1998030685A2 | 1998-07-16 | |||
| WO2000009680A1 | 2000-02-24 | |||
| WO2001007599A1 | 2001-02-01 |
Other References:
KAHN DAVID W ET AL: "Purification of plasmid DNA by tangential flow filtration", BIOTECHNOLOGY AND BIOENGINEERING. INCLUDING: SYMPOSIUM BIOTECHNOLOGY IN ENERGY PRODUCTION AND CONSERVATION, JOHN WILEY & SONS. NEW YORK, US, vol. 69, no. 1, 5 July 2000 (2000-07-05), pages 101 - 106, XP002151908, ISSN: 0006-3592
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