Table 3.
List of mRNA-LNP vaccine critical quality attributes related to stability and the analytical methods required to quantify these.
| Critical Quality Attribute | Analytical Methods | Ref. |
|---|---|---|
| RNA sequence integrity | Capillary Electrophoresis (CE), analytical high-performance liquid chromatography (HPLC), analytical ultra-high-performance liquid chromatography (UHPLC) | [55,56,57,58,59] |
| truncated RNA content | CE, analytical HPLC, analytical UHPLC | [55,56,57,58,59] |
| 5′ capped RNA percentage | Analytical HPCL, liquid chromatography-mass spectrometry (LC-MS), nuclease digestion followed by tandem mass spectrometry (MS/MS) quantitation | [25,29,56,60,61] |
| poly(A) tail length | LC-MS, reverse-phase HPLC and mass spectrometry (RP-HPLC-MS), CE | [29,55,62,63] |
| poly(A) tail level | Analytical HPLC, droplet digital polymerase chain reaction (ddPCR), MS | [29,56,64] |
| double-stranded RNA content | Immunoblotting, ELISA, RP-HPLC-MS | [27,29,57,65,66] |
| percentage encapsulated RNA | Absorbance assay, ribogreen assay, ion exchange HPLC, Raman spectroscopy, size-exclusion chromatography with multiangle light scattering (SEC-MALS) | [27,56,67,68] |
| LNP size | dynamic light scattering (DLS), nanoparticle tracking analysis, SEC-MALS | [27,56,59,67,68,69] |
| LNP polydispersity | DLS, nanoparticle tracking analysis, SEC-MALS | [27,56,59,67,68,69] |
| lipid-RNA adduct impurities | Ion pair RP-HPLC, HPLC, UPLC | [25,70] |