Supported binary liposome vesicle-gold nanoparticle for enhanced label free DNA and protein sensing

Supported binary liposome mixture of cationic liposome N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium propane (DOTAP) and the zwitterionic liposome 1,2-Dioleoyl-sn-Glycero-3-Phosphoethanolamine (DOPE) were tethered on thiol monolayers in the absence and presence of gold nanoparticle to enhance sensor stability and sensitivity for label free DNA and protein sensing for the first time. Cysteamine hydrochloride (Cyst), 3-Mercaptopropionic acid (MPA), 11-Mercaptoundecanoic acid (MUDA) and 11-amino-1-undecane thiol (AUT) monolayers were used as tethers on gold surfaces. Electrochemical studies in the presence of [Fe(CN)6]³⁻/⁴⁻ indicate that the presence of both DOPE and AuNP decreases the electrostatic interaction between DOTAP and MPA layer during the formation of DOPE-DOTAP-AuNP (DDA) whereas they enhance the repulsive force on the Cyst and AUT monolayers. In the thiol monolayer supported DDA, the gelation of neutral lipid DOPE by the AuNP is disfavored which inturn promotes stability of vesicle structure. The membrane protein melittin's interaction with the DDA indicates the presence of intact vesicle by showing decreased charge transfer for the MUDA and AUT in the presence of [Fe(CN)6]³⁻/⁴⁻. On the contrary, the presence of the bilayer and semi circled DDA on the MPA and cysteamine layers were confirmed by the increased redox reaction. Atomic Force Microscopic (AFM) and Transmission Electron Microscopic (TEM) images support the presence of an array like semi circled DDA on the MPA and well separated DDA vesicles on the MUDA with variable sizes. Dynamic Light Scattering (DLS) and Fourier Transform Infrared spectroscopy (FTIR) suggest effective coordination between DOPE, DOTAP and AuNP. Label free DNA hybridization sensing in presence of the negatively charged [Fe(CN)6]³⁻/⁴⁻ indicates the lowest DNA detection limit of 1×10⁻¹⁴M with linearity range 1×10–¹³ to 1×10⁻⁹M. Similarly, streptavidin sensing shows the lowest detection of 1ngml⁻¹ with a linear range 100ng to 1µg due to the increased reactive sites and distance.