A Novel Method of Immobilizing Antigens on Gold Electrode for Immunosensing

Nguyen Hoang Phuong Uyen, Gajovic-Eichelmann Nenad, Frank. F. Bier, Ngo Vo Ke Thanh

Abstract


An antigen modified gold electrode based on a self-assembled lipoic acid monolayer has been developed. The contact angle measurement and cyclic voltammetry confirm the formation of a dense self-assembled monolayer on gold from 100 mM lipoic acid in ethanol. The electrochemical behavior shows a stable activity in a range of potential from -0.2 V to 0.65 V vs. Ag/AgCl. By applying a potential of 1 V vs. Ag/AgCl the monolayer is completely removed by oxidative desorption and a clean gold surface re-established. This allows for an easy renewal of the gold surface and recycling of modified immunosensor chips. This lipoic acid monolayer was covalently functionalized with a small molecule antigen synthesized from progesterone-3-(O-carboxymethyl)oxime and 4,7,10-trioxa-1,13 tridecanediamine to form a well ordered, low unspecific binding, antigen layer for an antibody-antigen interaction study.The effectiveness of antigen - antibody binding reaction was demonstrated by fluorescence imaging using a fluorescence-labeled anti-progesterone antibody as the binding partner.
The new approach represents a way of improving the surface chemistry of electrochemical, surface-plasmon resonance and QCM-based immunosensors for which reusability, simplicity and sensitivity in flow-injection mode are required.

Keywords


antigen immobilization, electrochemical immunosensor, lipoic acid monolayer

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DOI: https://doi.org/10.15625/0868-3166/25/2/6239 Display counter: Abstract : 633 views. PDF : 57 views.

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