Processes of purifying oligonucleotides
AssigneeIsis Pharmaceuticals, Inc.
InventorsMax N. Moore, John Charles Arthur, Kent Vansooy, Anthony N. Scozzari
Life of this patent
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Abstract
Methods for preparing purified oligonucleotides by treating a solution comprising an oligonucleotide with an aggregating agent and a precipitation enhancer under conditions sufficient to form an oligonucleotide aggregate and isolating the oligonucleotide aggregate to produce a purified oligonucleotide.
Claims61
A method for preparing a purified oligonucleotide comprising the steps of:
- a) providing a solution comprising an oligonucleotide, said solution being effluent obtained from chromatography of crude oligonucleotide;
- b) treating said solution with an aggregating agent and a precipitation enhancer under conditions sufficient to form an oligonucleotide aggregate;
- and
- c) isolating said oligonucleotide aggregate to form said purified oligonucleotide.
A method for preparing a purified oligonucleotide comprising the steps of:
- a) providing a solution comprising an oligonucleotide, said solution being effluent obtained from chromatography of crude oligonucleotide;
- b) treating said solution with ethanol, wherein said ethanol is at a temperature of between about 15° C. and about 25° C. and a sodium salt under conditions sufficient to form an oligonucleotide aggregate;
- and
- c) isolating said oligonucleotide aggregate to form said purified oligonucleotide.
A method for preparing a purified oligonucleotide comprising the steps of:
- a) providing a solution comprising an oligonucleotide, said solution being effluent obtained from chromatography of crude oligonucleotide;
- b) treating said solution with a precipitation enhancer with subsequent treatment with an aggregating agent under conditions sufficient to form an oligonucleotide aggregate;
- and
- c) isolating said oligonucleotide aggregate to form said purified oligonucleotide.
A method for preparing a purified oligonucleotide comprising the steps of:
- treating a first solution comprising a 5′-protected oligonucleotide with an aggregating agent under conditions sufficient to form a first oligonucleotide aggregate;
- isolating said first oligonucleotide aggregate;
- dissolving the isolated first oligonucleotide aggregate in a solvent thereby forming a second solution;
- treating said second solution with a deprotecting reagent effective to remove said 5′-protecting groups;
- treating said second solution with an aggregating agent and a precipitation enhancer under conditions sufficient to form a second oligonucleotide aggregate;
- isolating said second oligonucleotide aggregate;
- dissolving said second oligonucleotide aggregate in a solvent to give a third solution;
- and
- treating said third solution with an aggregating agent and a precipitation enhancer under conditions sufficient to form a third oligonucleotide aggregate;
- and
- isolating said third oligonucleotide aggregate to give said purified oligonucleotide.
A method for preparing a purified oligonucleotide comprising the steps of:
- treating a first solution comprising an oligonucleotide with an aggregating agent and a precipitation enhancer under conditions sufficient to form a first oligonucleotide aggregate;
- isolating said first oligonucleotide aggregate;
- dissolving the isolated first oligonucleotide aggregate in a solvent thereby forming a second solution;
- treating said second solution with an aggregating agent and a precipitation enhancer under conditions sufficient to form a second oligonucleotide aggregate;
- and
- isolating said second oligonucleotide aggregate to give said purified oligonucleotide.
Dependencies are inferred from the claim text.