Nanobioelectrochemical Methods for the Investigation of Bioafinity Interactions

Victoria V. Shumyantseva, Institute of Biomedical Chemistry, Russian Academy of Medical Science, Moscow

Abstract: Nanometer-sized materials have attracted broad attention, and the fabrication of nanostructured electrodes has become an important part for electrochemical investigations. Gold nanoparticles are the most stable metal nanoparticles with size-related electronic and optical properties. Biomaterials such as enzymes, antibodies/antigens, aptamers/antigens exhibit nanometric sizes, comparable to those of nanoparticles. The integration of biomolecules with nanosized materials yield hybrid materials that have catalytic or recognition properties. Analysis of electrochemical parameters of biomolecules-nanoparticle hybrid systems permits to use them for bioelectronic applications. Nanostructured electrodes permit to use direct methods for analysis of bioaffinity interactions.

An approach based on volt-amperes' characteristics of voltammograms and amperograms for the analysis of substrate specificity of cytochromes P450 was proposed. To obtain nanoelectrode systems, based on gold nanoparticles on the electrode surface we used colloidal Au solution stabilized with lyotropic liquid crystalline phase of membrane-like synthetic surfactant didodecyldimethylammonium bromide DDAB. The electrochemical behavior of nanostructured electrodes with immobilized cytochromes P450 2B4, 1A2, 3A4, 11A1 (P450scc), and 51b1 (sterol 14a-demethylase, CYP51b1) in the presence of typical substrates and inhibitors of these forms were studied. Based on the results of amperometry, cyclic voltammetry, voltammetric analysis (DPV and SWV) and intermittent pulse amperometry (IPA), it was possible to conduct a search and to study the kinetic parameters of potential substrates and inhibitors of cytochrome P450. The proposed electrochemical approach is a sort of a bio-bar code of cytochrome P450 for the determination of the substrate/inhibitor P450's competence. The method of electro analysis may be applied to creation of multichannel electrochemical plates (chips, panels) with immobilized cytochromes P450.

Electrochemical immunoassays for antibodies to myoglobin/myoglobin interaction were investigated with voltammetric analysis. The detection limit of myoglobin detection was 175 ng/ml (10 nM).

About the speaker: Victoria Shumyantseva is the head of the Nanobioelectrochemistry Laboratory at the Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow, Russia. She received her Ph.D. degree in Chemistry of Nucleic Acids from the Department of Chemistry of Moscow State University, and her Dr.Sci. degree in Biochemistry from the Institute of Biomedical Chemistry. Her research interests include investigation of hemoproteins of medical importance, especially cytochrome P450s, electrochemistry of cytochrome P450s, nanocomposite materials and nanoscale systems for bioelectrochemical investigations, electroanalytical methods for the investigation of bioaffinity interactions. She has authored over 60 scientific publications. Since 2007, she is an Associate Professor at the Biochemistry Department of Moscow Medicinal University where she teaches a course on physico-chemical enzymology.

 

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