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Related Experiment Videos

Quantitative analysis of modified antisense oligonucleotides in biological fluids using cationic nanoparticles for

M Gerster1, J Schewitz, H Fritz

  • 1Research Center for Nucleic Acid and Peptide Chemistry, University of Tübingen, Auf der Morgenstelle 18, Tübingen, D-72076, Germany.

Analytical Biochemistry
|September 29, 1998
PubMed
Summary

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A new solid-phase extraction method using cationic polystyrene nanoparticles effectively quantitates modified antisense oligonucleotides in human plasma and urine. This technique offers high accuracy and recovery, even at low nanomolar concentrations.

Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Nanotechnology

Background:

  • Antisense oligonucleotides (ASOs) require robust analytical methods for accurate quantitation.
  • Existing extraction methods may face limitations with modified ASOs in complex biological matrices.
  • Solid-phase extraction (SPE) offers a promising approach for ASO isolation.

Purpose of the Study:

  • To investigate a novel solid-phase extraction (SPE) method using cationic polystyrene nanoparticles for ASO quantitation.
  • To assess the suitability of this SPE method for both terminally and backbone-modified ASOs.
  • To evaluate the extraction efficiency in human plasma and urine.

Main Methods:

  • Solid-phase extraction utilizing cationic polystyrene nanoparticles.

Related Experiment Videos

  • Quantitative analysis via capillary gel electrophoresis.
  • Extraction from human plasma and urine matrices.
  • Investigation of linearity, accuracy, and recovery rates.

    • Main Results:

    • High linearity and accuracy were demonstrated for modified ASOs, including oligodeoxyribonucleotide-palmityl conjugates and modified oligoribonucleotides.
    • Optimized conditions yielded high recovery rates (up to 95%) and purity for ASOs down to 5 nM concentrations.
    • No significant differences in recovery were observed between human plasma and urine.
    • Selective recovery of phosphodiester and phosphorothioate oligonucleotides was noted when nanoparticle capacity was exceeded, suggesting modification-dependent enrichment potential.

    Conclusions:

    • The developed SPE method using cationic polystyrene nanoparticles is suitable for accurate quantitation of modified antisense oligonucleotides in biological fluids.
    • The method provides high yields and sensitivity, applicable to low nanomolar concentrations.
    • The observed selective recovery offers a valuable tool for modification-dependent enrichment of ASOs from complex mixtures.