A facile entrapment approach to construct PEGylated polyplexes for improving stability in physiological condition

PEGylated polyplexes had been proved to improve the stability of DNA complexes. However, the conjugation reaction might reduce the capacity of efficient DNA complexation. Herein we described an easy and favorable approach to construct PEGylated polyplexes via entrapping poly(ethylene glycol) cholesterol ether (CPEG) into polyplexes. It was of interest to find the addition sequence of CPEG had great effect on the stability of polyplexes in physiological salt concentration. The addition of CPEG into the formed PEI(25k)/DNA polyplexes had no effect to improve the stability. Whereas by the "CPEG first" method of adding CPEG and PEI(25k) mixture into the DNA solution, the PEI(25k)/CPEG/DNA polyplexes showed excellent anti-aggregation effect and enhanced transfection efficiency in physiological condition. The difference performance might be explained by the possibility of CPEG entrapment. By the "CPEG first" method, PEGylated polyplexes was constructed due to the hydrophobic interaction between the cholesterol group of CPEG and hydrophobic charged-compensated core. The PEG coating significantly improved the stability of polyplexes in physiological condition. This facile entrapment approach to prepare PEGylated polyplexes might have great potential in non-viral gene delivery research and application.