Table 3.
Test methods used for ASOs, siRNAs, and mRNAs.
| Category | Tests | ASOs | siRNAs | mRNAs |
|---|---|---|---|---|
| Active substance | Identification | Duplex melting temperature (UV absorbance) | Circular dichroism | Capillary gel electrophoresis |
| FTIR | Duplex melting temperature (UV absorbance) | RT-Sanger sequencing | ||
| IP-RPLC-UV-MS | FTIR | RT-PCR | ||
| MS-MS | IP-RPLC UV-MS | |||
| NMR spectroscopy (1H, 13C and 31P) | MS-MS | |||
| X-ray diffraction | NMR spectroscopy (1H, 13C and 31P) | |||
| SEC-UV | ||||
| UV spectroscopy | ||||
| Assay | IP-RPLC-UV-MS | AEX-UV | UV spectroscopy | |
| UV spectroscopy | ||||
| Impurities | IP-RPLC-UV-MS | AEX -UV | ddPCR | |
| IP-RPLC-UV-MS | Immunoblot | |||
| SEC-UV | IP-RPLC | |||
| 2D-LC (AEX -UV (first dimension) and IP-RPLC-MS (second dimension)) | qPCR | |||
| Finished medicinal product | Identification | Duplex melting temperature (UV spectroscopy) | AEX-UV | Capillary gel electrophoresis |
| IP-RPLC-UV-MS | Duplex melting temperature (UV spectroscopy) | RT-Sanger sequencing | ||
| FTIR | ||||
| IP-RPLC-UV-MS | ||||
| Assay | IP-RPLC-UV-MS | AEX-UV | AEX-UV | |
| UV spectroscopy | Fluorescence assay | |||
| Impurities | IP-RPLC-UV-MS | AEX-UV | IP-RPLC-UV-MS | |
| IP-RPLC-UV-MS |