Oligonucleotide Standards

Synthetic oligonucleotides are short nucleic acid chains that can control gene expression in a sequence-specific manner. They can target different diseases by interfering with RNA functions at the cellular level. As a new class of therapeutics, synthetic oligonucleotides have shown great promise in the treatment of neurological disorders, metabolic diseases, cancers, and other diseases.

There is limited published guidance from regulatory agencies on quality control of oligonucleotide drugs, including control of raw materials or product-related impurities. USP now offers standards to support quality assessment of critical raw materials used in oligonucleotide synthesis. These standards also help you navigate the current regulatory environment around assessing product-related impurities that can affect safety and efficacy.

Reference standards for raw material testing used in oligonucleotide manufacturing

USP DNA Phosphoramidite Reference Standards

USP’s Oligonucleotide Educational Resources

Phosphoramidites are modified nucleosides used in the chemical synthesis of oligonucleotides. Synthetic oligonucleotides often contain low levels of multiple classes of impurities which can ultimately influence their efficacy and toxicity.

USP has developed DNA phosphoramidites reference standards that can help manufacturers identify and control impurity profiles during oligonucleotide synthesis using various analytical techniques, including HPLC, 31P NMR, GC, and LC-MS. Ultimately these standards may help yield better control over manufacturing process.

The DNA phosphoramidites reference standards can support the quality assessment of raw materials throughout the life cycle from oligonucleotide products development to lot release.

USP DNA Phosphoramidite Reference Standards

Each USP DNA Phosphoramidite Reference Standard is characterized by:

  1. HPLC analysis, demonstrating isomer retention times and product purity.
  2. 31P NMR spectroscopy, illustrating compound identification (signals for right-handed (Rp) and left-handed (Sp) diastereoisomers) and purity assignment.
  1. Therapeutic Oligonucleotides Production: CMC – Regulatory – Quality
  2. Developing Product-specific Guidances on Oligonucleotides for Generic Drug Development
  3. USP Standards to Support Quality of Peptide and Oligonucleotide Therapies
  4. Strategies for the Control of Impurities in Oligonucleotide Synthesis
  5. Terminal Sterilisation of Oligonucleotide Drug Products
  6. In-depth Impurity Profiling of Synthetic Oligonucleotides by High Resolution Mass Spectrometry
  7. CMC and Regulatory Challenges for Oligonucleotide Drugs
  8. Evaluation of Chromatographic Techniques for Oligonucleotide Separation and Subsequent Detection by HRMS for Routine Testing
  9. Applications of Green Chemistry in Oligonucleotide Manufacturing
  10. CMC Regulatory Considerations for Oligonucleotides and Peptides: Similarities and Differences
  11. Application of NMR Spectroscopy for Characterization and Identity Testing of Oligonucleotide Drug Molecules
  12. Antisense Oligonucleotide Specification and Release Tests using a Risk-Based Platform Approach
  13. Method Development for the Characterization of Synthetic Oligonucleotides by LC-MS
  14.  Analytical Method Development for Critical Impurities in Phosphoramidites