The development of DNA delivery system based on ionic liquid and carboxylic acid modified polyethyleneimine

Recombinant DNA vaccines offer significant potential for disease prevention and therapy, but their clinical success is often limited by poor immunogenicity, low cellular uptake, instability, and inefficient delivery without proper carriers. To address these challenges, we developed a series of novel cationic delivery systems by modifying branched polyethyleneimine (PEI, 25 kDa) with biocompatible carboxylic acids either lactic acid or glycolic acid and incorporating choline-based ionic liquids (choline glycolate [CG] and choline lactate [CL]) to create advanced mPEI/ionic liquid (IL) formulations. These systems were designed to enhance DNA complexation, protect against enzymatic degradation, improve nanoparticle stability, and fine-tune physicochemical properties for optimal cellular interaction. The resulting polyplexes formed stable nanoparticles with diameters 100 –125 nm and surface charges of +24 to +29 mV, supporting efficient cellular uptake. Compared to unmodified PEI, the modified formulations showed markedly reduced cytotoxicity and significantly improved transfection performance. Among all tested combinations, the combination of lactic acid–modified PEI with choline glycolate (LA-mPEI + CG) was the most effective, achieving a 76% increase in transfection efficiency, a 66% improvement in cellular uptake, and 66% enhanced in cell viability. These findings highlight the synergistic advantage of combining carboxylic acid modification PEI with ionic liquid incorporation, providing a promising strategy for safer and more effective non-viral DNA delivery. This platform may serve as a foundation for future advancements in gene therapy and DNA vaccine development.