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Related Experiment Videos

Purification of antisense oligonucleotides.

R R Deshmukh1, D L Cole, Y S Sanghvi

  • 1Manufacturing Process Department, ISIS Pharmaceuticals, Inc., Carlsbad, California 92008, USA.

Methods in Enzymology
|December 14, 1999
PubMed
Summary

Reversed-phase (RP) and anion-exchange (AX) chromatography effectively purify synthetic oligonucleotides for drug therapy. AX chromatography offers scalable, cost-effective purification with reduced solvent use for large-scale oligonucleotide drug production.

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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.

Related Experiment Videos

  • 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.