Electrically addressed covalent immobilization of biomolecules to the defined electrodes of an electrode array is described. It is based on Michael addition of the thiol group of biomolecules to α,β-unsaturated carbonyl groups of benzoquinone. This "click" reaction was tested by immobilization of a number of thiolated compounds on the simplest array consisting of two gold electrodes coated by a self-assembled monolayer of benzoquinone-terminated hexanethiol. Electrically controlled binding of hexanethiol, ferrocenylhexanethiol, human serum albumin and thiol-terminated single-stranded DNA (ssDNA) was investigated. The binding was studied using cyclic voltammetry, X-ray photoelectron spectroscopy and surface plasmon resonance. The reaction requires the oxidized state of the benzoquinone moiety; this can be reached by applying of a moderate anodic potential to the electrode. Surface plasmon resonance measurements demonstrated that the thiol-modified ssDNA immobilized by this technique binds complementary synthetic oligonucleotides or PCR-amplified DNA fragments. The developed technology of electrical addressing of covalent immobilization can be applied for fabrication of sensor arrays.
Keywords: Addressable immobilization; Cyclic voltammetry; Immobilization of biomolecules; Michael addition; Self-assembled monolayer; Surface plasmon resonance.
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