Systematic Approach to the Development of Microfabricated Biosensors: Relationship between Gold Surface Pretreatment and Thiolated Molecule Binding.

Despite the increasing popularity of microfabricated biosensors due to advances in technologic and surface functionalization strategies, their successful implementation is partially inhibited by the lack of consistency in their analytical characteristics. One of the main causes for the discrepancies is the absence of a systematic and comprehensive approach to surface functionalization. In this article microfabricated gold electrodes aimed at biosensor development have been systematically characterized in terms of surface pretreatment, thiolated molecule binding, and reproducibility by means of X-ray photoelectron scattering (XPS) and cyclic voltammetry (CV). It has been shown that after SU-8 photolithography gold surfaces were markedly contaminated, which decreased the effective surface area and surface coverage of a model molecule mercaptohexanol (MCH). Three surface pretreatment methods compatible with microfabricated devices were compared. The investigated methods were (i) cyclic voltammetry in dilute H2SO4, (ii) gentle basic piranha followed by linear sweep voltammetry in dilute KOH, and (iii) oxygen plasma treatment followed by incubation in ethanol. It was shown that all three methods significantly decreased the contamination and increased MCH surface coverage. Most importantly, it was also revealed that surface pretreatments may induce structural changes to the gold surfaces. Accordingly, these alterations influence the characteristics of MCH functionalization.