Function of hexameric RNA in packaging of bacteriophage phi 29 DNA in vitro

A methodology based on high-performance gel exclusion chromatography (HPLC-GEC) has been developed to perform permeability studies of vesicles. Encapsulation of two marker isothiocyanate fluorescein (FITC) dextrans of 4400 and 40,500 molecular mass was used as a model system. Combination of two TSK-PW columns, one efficient in vesicle sizing (G6000 PW), the other in that of dextrans (G4000 PW), was required to achieve complete particle separation and to remove entirely the unentrapped dextran after encapsulation into vesicles. Coupling fluorescence and light scattering detection allowed to control the efficiency of the separation, to quantify the vesicle leakage and to follow both the integrity of the vesicles and changes in their size. This methodology can be applied to other fields such as encapsulation of water soluble compounds and drug delivery systems.