Electrochemical and spectroscopic study of octadecyltrimethylammonium bromide/DNA surfoplexes.
The use of cationic micelles consisting of octadecyltrimethylammonium bromide (C18TAB) to compact calf thymus DNA has been investigated in aqueous buffered solution at 310.15 K by means of conductometry, electrophoretic mobility, and several fluorescence spectroscopy methods. The results indicate that C18TAB micelles, consisting of 44 monomers on average, may compact DNA molecule by an electrostatic interaction that takes place at the cationic spherical micelle surface. The surfoplexes thus formed show a surface density charge that goes from negative to positive values at a Lmic/D mass ratio of around 1.0 (where Lmic and D are the masses of micellized cationic surfactant and DNA), called the isoneutrality ratio (Lmic/D)phi. Values of this characteristic parameter, determined in this work not only from the electrochemical experimental data but also from spectroscopic measurements, are in very good agreement with those ones calculated from molecular parameters and some other properties also obtained in this work. The electrostatic character of the DNA-micelle interaction has been confirmed by analyzing the decrease in fluorescence emission of the fluorophore ethidium bromide, EtBr, initially intercalated between DNA base pairs, as long as the surfoplexes are formed. Fluorescence anisotropy experiments have revealed that micelle packing becomes more rigid in the presence of DNA, but once the surfoplex is formed, the fluidity increases with the Lmic/D mass ratio, attaining its maximum when the isoneutrality ratio is exceeded. This fact, together with the net positive charge of the surfoplexes with the Lmic/D mass ratio over the isoneutrality ratio, makes this regimen of lipid and DNA content the optimum for efficiency in the transfection process.
Departamento de Química Física I, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, 28040-Madrid, Spain.
Alberto
A
Rodríguez-Pulido
Emilio
E
Aicart
Elena
E
Junquera
D000473:Alkanes / Q000737:chemistry*; D016880:Anisotropy; D001671:Biochemistry / Q000379:methods; D002412:Cations; D003102:Colloids / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D004553:Electric Conductivity; D004563:Electrochemistry / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D008055:Lipids / Q000737:chemistry; D008823:Micelles; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013053:Spectrophotometry / Q000379:methods*; D013499:Surface Properties; D013696:Temperature
D000473:Alkanes; D002412:Cations; D003102:Colloids; D008055:Lipids; D008823:Micelles; D000644:Quaternary Ammonium Compounds; C518260:octadecyltrimethylammonium bromide; D004247:DNA