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US 6,469,158 B1Granted
Synthesis, deprotection, analysis and purification of RNA and ribozymes
AssigneeRibozyme Pharmaceuticals, Inc.
InventorsNassim Usman, Francine Wincott, David Sweedler, Leonid Beigelman, Lech W. Dudycz, Susan Grimm, Anthony DiRenzo, Danuta Tracz
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Abstract
Method for purification and synthesis of RNA molecules and enzymatic RNA molecules in enzymatically active form.
Claims17
A method for the purification of an enzymatic RNA molecule of 28 or more nucleotide bases comprising the steps of:
- a) loading a mixture comprising said RNA on to an anion exchange high-performance liquid chromatography (HPLC) column;
- b) eluting said RNA by passing a buffer through said column;
- and
- c) collecting the eluate from said column and recovering said RNA from said eluate;
- wherein said method is performed under conditions which allow for the purification of an enzymatic RNA molecule of 28 or more bases.
2 more elements
A method for the purification of an enzymatic RNA molecule of 28 or more nucleotide bases comprising the steps of:
- a) loading a mixture comprising said RNA on to reverse phase high-performance liquid chromatography (HPLC) column, wherein said RNA comprises a 5′-protecting group;
- b) eluting said RNA by passing a buffer through said reverse phase column;
- c) removing the 5′-protecting group from said RNA;
- d) loading said RNA lacking said 5′-protecting group onto an anion exchange high-performance liquid chromatography (HPLC) column;
- e) eluting said RNA by passing a buffer through said anion exchange column;
- and
- f) collecting the eluate from said anion exchange column and recovering said RNA from said eluate;
- wherein said method is performed under conditions which allow for the purification of an enzymatic RNA molecule of 28 or more bases.
5 more elements
Dependencies are inferred from the claim text.