Patent ID: 6989442
Filing Date: 2006-01-24
Classification: C07H,C12N

Abstract:
1. A process for synthesis, one-pot deprotection, and purification of a nucleic acid molecule having one or more ribonucleotides, comprising the steps of: a) synthesizing said nucleic acid molecule using a method selected from the group consisting of solid phase phosphoramidite, solution phase phosphoramidite, solid phase H-phosphonate, solution phase H-phosphonate, hybrid phase phosphoramidite, and hybrid phase H-phosphonate-based synthetic methods; b) contacting said nucleic acid molecule from step (a) with aqueous alkylamine, trialkylamine, or a mixture of alkylamine and trialkylamine, under conditions suitable for the removal of any 2′-amino protecting groups, exocyclic amino (base) protecting groups and/or phosphate protecting groups, which may be individually present or absent, from said molecule; c) contacting reaction mixture having said nucleic acid molecule from step (b) with a mixture of polar solvent and trialkylamineehydrogen fluoride under conditions suitable for the removal of a 2′-OH protecting group; d) loading reaction mixture having said nucleic acid molecule from step c) onto ion-exchange purification media in a suitable buffer comprising buffers selected from the group consisting of water, 20% ethanol in about 20 mM sodium phosphate and about 0.1 M NaCl and acetonitrile in about 20 mM sodium phosphate and about 0.1 M NaCl; e) applying a purification gradient using a suitable elution buffer, analyzing the fractions and allowing for the Pure fractions to be pooled and desalted.