A series of linear polymers containing varying amounts of ethylenimine or N-propylethylenimine unitswere synthesized by hydrolysis and/or reduction of polyethyloxazolines. The pKas of the polyamineswere determined potentiometrically. Gel mobility shift assay showed that the efficiency of DNAcomplexation was related to the fraction of amino groups that are protonated at neutral pH. Theeffects of cationic charge density and molar weight of the polymers on the transfection efficiency wereevaluated on HepG2 cells. The results obtained with different copolymers show that the transfectionefficiency primarily depends on the fraction of ethylenimine units included in the polymer albeit themolar weight is also of importance. On the basis of the results obtained with poly(N-propylethylenimines), we also demonstrate that the high transfection efficiency of polyethylenimines does not solelyrely on their capacity to capture protons which are transferred into the endo-lysosomes duringacidification.