Purification of antisense oligonucleotides

Area of Science:

• Biochemistry
• Chemical Engineering
• Pharmaceutical Sciences

Background:

• Synthetic oligonucleotides are crucial for therapeutic applications like antisense drug therapy.
• Chromatographic techniques are essential for achieving the high purity required for these applications.
• Reversed-phase (RP) and anion-exchange (AX) chromatography are established methods for oligonucleotide purification.

Purpose of the Study:

• To compare the effectiveness and scalability of RP and AX chromatography for synthetic oligonucleotide purification.
• To evaluate the advantages and limitations of each technique for potential large-scale pharmaceutical production.
• To inform the selection of optimal purification strategies for oligonucleotide-based therapeutics.

Main Methods:

• Utilized reversed-phase high-performance liquid chromatography (RP-HPLC) with hydrophobic-protecting groups.
• Employed anion-exchange high-performance liquid chromatography (AX-HPLC) for oligonucleotide purification.
• Assessed purification performance across various scales, from small to potentially ton-scale production.

Main Results:

• RP chromatography excels in purifying diverse modified oligonucleotides and is readily scalable with minimal process development.
• AX-HPLC provides high-purity products with higher media capacities, though method development is more intensive.
• While AX yield per operation may be lower, isolated yields can be comparable or superior to RP-HPLC, with potential for reduced costs and solvent usage at scale.

Conclusions:

• Both RP and AX chromatography are viable for high-purity oligonucleotide purification.
• AX-HPLC presents a scalable and potentially more economical option for ton-scale production of oligonucleotide drugs, with reduced organic solvent needs.
• Further method development for AX-HPLC can optimize its application for the growing oligonucleotide therapeutics market.