The integrity and properties of mycolic acid (MA) antigens integrated into a self-assembled monolayer (SAM) of N-(2-mercaptoethyl)octadecanamide, (MEODA), on a gold electrode have been interrogated using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). EIS data showed that Au–MEODA and Au–MEODA–MA behave as microelectrode arrays, with pinholes acting as the microelectrodes that permit electron transport between a redox-active probe in solution and the underlying gold surface. The average radii of the pinholes (ra) and half the distance between the centers of the neighbouring pinholes (rb), were estimated from EIS using the pore size model, and discussed. Anti-MA antibodies present in a tuberculosis (TB)-infected patient (co-infected with HIV) strongly interact with Au–MEODA–MA showing a rather compact and stable bio-complex structure that is virtually defect-free. The electrochemical impedimetric properties associated with the ability of the Au–MEODA–MA to discriminate between TB positive and negative human sera are also discussed. We prove that the Au–MEODA and Au–MEODA–MA electrodes, as well as the MA-anti-MA antibody interactions, are characterized with time-constant dispersion, typical of microstructures with grain/grain boundary phases. These crucial physico-electrochemical insights into the behaviour of surface-confined MA should provide a useful basis for the design and development of a potential impedimetric immunosensing platform for active tuberculosis.
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