C - Chemistry – Metallurgy – 07 – H
Patent
C - Chemistry, Metallurgy
07
H
C07H 21/00 (2006.01) C12P 19/34 (2006.01)
Patent
CA 2361079
A method for purifying an oligonucleotide that comprises providing an oligonucleotide attached to a substrate, wherein the oligonucleotide contains phosphate protecting groups; contacting the oligonucleotide with a reagent, e.g., an organic amine, that cleaves the phosphate protecting groups from the oligonucleotide without detaching the oligonucleotide from the substrate; isolating the oligonucleotide attached to the substrate from the cleaved phosphate protecting groups; and cleaving the oligonucleotide from the substrate. This method provides crude oligonucleotide mixtures that are easier to purify and from which the desired full-length oligonucleotide product can be isolated in higher yields.
L'invention concerne un procédé de purification d'un oligonucléotide, qui consiste à fournir un oligonucléotide fixé à un substrat, l'oligonucléotide contenant des groupes de protection phosphatés, à mettre en contact l'oligonucléotide avec un réactif, une amine organique, par exemple, qui produit le clivage des groupes de protection phosphatés de l'oligonucléotide sans détacher ce dernier du substrat, à isoler l'oligonucléotide fixé au substrat des groupes de protection phosphatés ayant subi le clivage, et à détacher l'oligonucléotide de son substrat par clivage. Le procédé fournit des mélanges d'oligonucléotides bruts plus faciles à purifier et desquels le produit nucléotidique complet voulu peut être isolé, à des rendements plus élevés.
Bhan Anila
Holmberg Lars
Amersham Pharmacia Biotech Inc.
Fetherstonhaugh & Co.
Ge Healthcare Bio-Sciences Corp.
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