Process for the preparation of endotoxin-free or endotoxin-depleted nucleic acids and/or oligonucleotides for gene therapy

US6297371A · kind A · utility

Assignee

• QIAGEN GmbH · DE

Inventors

• Metin Colpan · Essen, DE
• Joachim Schorr · Düsseldorf, DE
• Peter Moritz · Wiesendangen, CH

Key dates

• Filing date: Feb 22, 1999
• Grant date: Oct 2, 2001
• Expiry date: Feb 22, 2019

Classification

• Technology area (CPC Y): Emerging Cross-Sectional Technologies
• CPC primary: Y10T436/10
• WIPO field: Chemical engineering
• WIPO sector: Chemistry

Abstract

A process for the isolation and purification of nucleic acids and/or oligonucleotides for use in gene therapy wherein said nucleic acids and/or oligonucleotides are isolated or purified from an essentially biological source, characterized in that PA1 said essentially biological sources are lysed, the fractions obtained are optionally freed or depleted from the remainder of said biological sources by per se known mechanical methods, such as centrifugation, filtration; PA1 the fractions thus treated are subsequently treated with affinity chromatographic material or with inorganic chromatographic material for the removal of endotoxins; followed by PA1 isolation of said nucleic acids and/or oligonucleotides on an anion exchanger which is designed such that DNA begins to desorb from the anion exchanger only at an ionic strength corresponding to a sodium chloride solution of a concentration higher by at least 100 mM than one corresponding to the ionic strength at which RNA begins to desorb from the anion exchanger material.