In order to develop a plat surface for the printing anti – E.coli O157:H7 polyclonal antibody used in microarray technology and immunosensors, the substrates are coated with thick layer of gold and are functionalized with self-assembled monolayer technique. In experiment are two types of substrates: one is formatted with silicon substrate and other is formatted with polymer substrate. One of the most important points in the design of an immunosensor and a microarray is the choice of immobilization method. Recently, self –assembled monolayers (SAMs) generated by adsorption of alcanethiols on gold have been the subject of extensive research. These techniques will allow the study of the adsorption of proteins to different SAMs formed on the gold surfaces. In this work, the cyclic voltammetry and spectroscopy technique (Spectroscopic Ellipsometry, FTIR, Raman) have been utilized for surfaces characterization. The vibration molecule frequencies of the thiol monolayer can be detected with infrared spectroscopy (FTIR-ATR). Activation method included NHS/EDC for coupled SAM with an affinity purified polyclonal antibody to E. coli O157:H7 made in Goat. For microarray detection the anti-E. coli O157:H7 polyclonal antibody was labeled with fluorescein isothiocyanate (FITC).

The samples for microarray were prepared using anti-E. coli polyclonal antibody (KPL) in PBS (Sigma) in serial dilutions to determine optimal working concentrations. In the present study we report two substrates, silicon and polymer who are functionalized with different SAMs in order to immobilize efficient the anti-E O157:H7 polyclonal purified antibody.

This experimental study was revealed the necessity to investigate each step taking into account in the same time the factors that influence the chemistry of the surface and the further interactions as well.

The realisation of an e-coli sensor is envisaged and the comparison between the two substrates is achieved.

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