Purification of antisense oligonucleotides.
Chromatography is an effective tool for obtaining high-purity synthetic oligonucleotides for a variety of end uses, including antisense drug therapy. Reversed-phase and anion-exchange chromatographies are widely used techniques for this application. While selectivity of these techniques can be modified by methods such as ion-pair RP-HPLC or affinity chromatography, these are presently used only at small scales. RP chromatography makes use of terminal hydrophobic-protecting groups to increase retention and selectivity. The main advantages of the RP method are its utility for the purification of a wide variety of modified oligonucleotide structures, its applicability across a range of terminal hydrophobic groups, such as fluorescein, and its ready use from small scale to very large scale with a minimal requirement for process development. AX-HPLC can also give high-purity products at generally higher media capacities. A more extensive method development effort is typically required for the AX-HPLC purification of AO. The AX yield per unit operation can be lower, but the isolated yield of DMT-off desalted oligonucleotide can be equal to or higher than that from RP-HPLC. As additional AO drugs enter and mature in the market, there will be a potential need for ton-scale purification processes. AX provides a way to scale up production on somewhat less expensive equipment with reduced organic solvent requirements.
Deshmukh
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RR
Manufacturing Process Department, ISIS Pharmaceuticals, Inc., Carlsbad, California 92008, USA.
Cole
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DL
Sanghvi
Y S
YS
D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007202:Indicators and Reagents; D020319:Oligodeoxyribonucleotides, Antisense / Q000302:isolation & purification*; D016376:Oligonucleotides, Antisense / Q000302:isolation & purification*; D012997:Solvents; D013873:Thionucleotides
D007202:Indicators and Reagents; D020319:Oligodeoxyribonucleotides, Antisense; D016376:Oligonucleotides, Antisense; D012997:Solvents; D013873:Thionucleotides