The fabrication of self-ordered semiconductor (TiO2) and noble metal (Au) QDs arrays was successfully achieved by advanced nonlithographic template based method, namely using nanoporous alumina template. The emphasis was placed on the successful preparation of QDs arrays with the desired size, homogeneous distribution and optical (especially fluorescence) properties. Titania and gold QDs characterization by SEM, EDX and fluorescence spectroscopy was performed in order to verify their surface topography, chemical composition and emission properties in UV/VIS range of spectra, respectively. The surface biofunctionalization of QDs was realized via simple physical adsorption of glutathione tripeptide, which makes these arrays suitable for potential biosensing application, mainly in optical and electrochemical detection of biomoleculesin vitro.