A Nanoplasmonic Assay of Oligonucleotide-Cargo Delivery from Cationic Lipoplexes

A powerful new biophysical model is reported to assay nanocarrier lipid membrane permeability. The approach employs a nanophotonic biophysical membrane model as an assay to study oligonucleotide escape from delivery vector following fusion with endosomal membrane that relies on plasmonic hotspots within the receptor well, below the membrane to follow cargo arrival. Through the combined use of surface enhanced Raman spectroscopy and fluorescence lifetime correlation spectroscopy (FLCS), the model enables identification of a lipoplex-mediated endosomal-escape mechanism facilitated by DOTAP-oligonucleotide interaction that dictates the rate of oligonucleotide release. This work reveals a hitherto unreported release mechanism as a complex multistep interplay between the oligonucleotide cargo and the target membrane, rather than a process based solely on lipid mixing at the fusing site as previously proposed. This substantiates the observations that lipid mixing is not necessarily followed by cargo release. The approach presents a new paradigm for assessment of vector delivery at model membranes that promises to have wide application within the drug delivery design application space. Overall, this plasmonic membrane model offers a potential solution to address persistent challenges in engineering the release mechanism of large therapeutic molecules from their nanocarrier, which is a major bottleneck in intracellular delivery.