ssDNA Encapsulation Method in Soybean-Based Nanoliposomes of the ARGOMULYO Variety

Abstract

This invention concerns a method for the formulation and encapsulation of single-stranded DNA (ssDNA) into phosphatidylcholine-based nanoliposomes from Argomulyo soybean variety. This method includes the formation of liposomes by thin-layer hydration technique using environmentally friendly solvents ethanol-aquadest, followed by the direct incubation of ssDNA into liposomes at various lipid:ssDNA mass ratios (0.5:1, 1:1, and 1.5:1). Separation of free ssDNA was carried out by centrifugation at 16,000 g for 20 minutes. The formulation results showed the highest encapsulation efficiency (>90%) at a 1:1 ratio, with the majority particle size distribution in the range of 150–300 nm based on DLS number distribution analysis, as well as physicochemical characteristics with a zeta potential of around –2 to –4 mV.This method produces a simple, efficient, and biocompatible non-viral nucleic acid delivery system, and can be widely applied for molecular research and the development of liposome-based genetic therapy.