Table 2.

Methods for LNP characterization.

Characterization criteria of LNPs	Implications	Assay methods
Particle size	To determine size of nanoparticles	Dynamic light scattering (DLS)
Polydispersity index (PDI)	To assess homogeneity and size distribution
Zeta potential	Gives idea about surface charge of nanoparticles
Morphology & structure	To evaluate structural characteristics on the LNPs	Cryogenic transmission electron microscopy (Cryo-TEM), SAXS (small -angle x-ray scattering)
Encapsulation%	To quantify the payload inside LNPs.	Ribogreen/Picogreen assay
Integrity of the payload	To detect stability and integrity of entrapped nucleic acid, following encapsulation	Gel electrophoresis
Gene expression efficiency	Indicates effectiveness of LNP to deliver payload	In vitro/in vivo transfection