Patent ID: 6649751
Filing Date: 2003-11-18
Classification: A61K,C07K,C12N,C12Q

Abstract:
A process for deprotecting and purifying chemically synthesized RNA having one or more chemical modifications, comprising:(a) deprotection, wherein said deprotection comprises removing any exocyclic protecting groups from said RNA molecule by contacting said RNA with an alkylamine or a mixture of ammonium hydroxide and alkylamine at a temperature of about 60-70 degrees centigrade for about 5-15 minutes, under conditions suitable for the removal of said exocyclic protecting groups, wherein said alkylamine is selected from the group consisting of ethylamine, propylamine and butylamine; and (b) purification, wherein said purification comprises: (i) loading said RNA on to reverse phase high-performance liquid chromatography (HPLC) column, wherein said RNA comprises a 5â€²-protecting group; (ii) eluting said RNA by passing a suitable buffer through said reverse phase column; (iii) removing the 5â€²-protecting group from said RNA; (iv) loading said RNA lacking said 5â€²-protecting group on to an anion exchange high-performance liquid chromatography (HPLC) column; (v) eluting said RNA by passing a suitable buffer through said anion exchange column; and (vi) collecting the eluate from said anion exchange column and recovering said RNA from said eluate, under conditions which allow for the purification of the RNA.