PURIFICATION OF OLIGODEOXYNUCLEOTIDE PHOSPHOROTHIOATES USING DEAE 5PW ANION ION-EXCHANGE CHROMATOGRAPHY AND HYDROPHOBIC INTERACTION CHROMATOGRAPHY

摘要

The present invention provides improved methods for separating and purifying oligonucleotide phosphorothioates. In certain aspects of the invention, oligonucleotide phosphorothioates are purified by a method comprising applying a mixture of the oligonucleotide to a column containing a DEAE-5PW anion ion-exchange chromatography resin and eluting with an elution buffer having a concentration of sodium chloride of from about 0-2M. In other aspects of the invention, oligonucleotide phosphorothioates are purified by a method comprising applying a mixture of the oligonucleotide to a column containing a phenyl-sepharose fast flow chromatography resin or a phenyl-5PW chromatography resin and eluting with an elution buffer substantially free of salts. The methods of the present invention provide the ability to purify ammoniacal solutions of oligonucleotides cheaply, quickly, and on a large, process scale.