Home / Small molecules / Integrated Services / Oligonucleotide Analytical
Oligonucleotide Analytical
The hard measurement is a sequence one base short.
Analytical development
A dedicated analytical team covers oligonucleotide analysis across all development phases from preclinical through commercial manufacturing, for both drug substance and drug product.
The team is co-located with the API process development team, which is what allows a method to be developed against the material as it is made rather than after it arrives.
Drug substance analytical services
- Analytical development for drug substance specific methods (purity, sequencing) and feasibility for oligonucleotide generic methods
- Forced degradation study with impurity qualification and quantitation
- Release testing and stability study under R&D conditions
- Reference standard and impurity marker qualification and retesting
- Physicochemical property study and structure elucidation
- Microbiological safety: endotoxin limit and quantitation testing, bioburden for microbial limit test, microbe identification and strain typing
Drug product analytical services
- Container-closure integrity testing, including quantitative vacuum decay and high-voltage leak detection and qualitative microbial challenge and tracer liquid methods
- Foreign and particulate matter observed in situ, verified by flow imaging and identified by IR, Raman or EDX, with osmolality
- Purity by LC-MS, including identification and quantitation by Q-TOF HRMS and quadrupole LC-MS, for hard-to-resolve parenteral impurity analysis
- Sterility and microorganism identification
Special capabilities
- Process-related impurity identification by 2D UPLC coupled with Q-TOF HRMS
- Identification by tandem LC-MS/MS for sequence confirmation
- Oligonucleotide backbone composition by 31P NMR
- Diastereomer fingerprinting by 31P NMR
Quality control
- Method qualification and validation for IND, NDA, and commercial use
- Transfer of GMP validated methods according to USP <1224>
- Full release testing across raw materials, reagents, intermediates, drug substance, and drug product
- Accelerated and long-term stability studies under ICH conditions
Case study: purity by LC-UV-HRMS
Where impurities co-elute with full-length product in HPLC-UV, mass spectrometric purity is used instead for qualification and quantitation — routinely for tRNA, sgRNA, PMO, PPMO, and ASO.
Case study: resolution on a 100-mer sgRNA
Our HPLC purity method for sgRNA has moved from separating n−8 and n+4 impurities to separating n−2 and n+1.
Case study: sequencing a PPMO
Determining a peptide-PMO sequence means sequencing both the peptide and the PMO components, and the conjugation between the two intermediates.
Case study: screening purity methods for resolution
Ion-exchange chromatography and reverse-phase HPLC are both used for purity and impurity analysis of a PMO product. The two orthogonal techniques give different impurity profiles, and screening methods systematically produces a purity method with more resolved impurity peaks.
Instrumentation
- Chromatography: UPLC, HPLC, GC, IC, 2D-UPLC, HPLC-CAD
- Mass spectrometry: Q-TOF, MALDI-TOF, LC-Orbitrap, LC-MS, LC-QQQ, 2D LC-MS, GC-MS
- Physicochemical testing: FT-IR, UV-Vis, XRPD, DVS, DSC, TGA, polarimetry, 400 and 600 MHz NMR, MEC, thermal melting, circular dichroism
- Elemental analysis: ICP-OES, ICP-MS, CHN elemental analysis