Enzyme-based optical biosensors, preparation

Similar optical biosensors have been prepared for many other analytes. For example, a cholesterol optical biosensor has been devised based on fluorescence quenching of an oxygen-sensitive dye that is coupled to consumption of oxygen resulting from the enzyme-catalyzed oxidation of cholesterol by the enzyme cholesterol oxidase. Serum bilirubin has been detected using bilirubin oxidase, coimmobilized with a ruthenium dye, on an optical fiber.The bilirubin sensor was reported to exhibit a lower detection limit of iO Xmol/L, a linear range up to 30mmol/L, and a typical reproducibihty of 3% (CV), certainly adequate for clinical application. 

In this part we will describe recent achievements in the development of biosensors based on DNA/RNA aptamers. These biosensors are usually prepared by immobilization of aptamer onto a solid support by various methods using chemisorption (aptamer is modified by thiol group) or by avidin-biotin technology (aptamer is modified by biotin) or by covalent attachment of amino group-labeled aptamer to a surface of self-assembly monolayer of 11-mercaptoundecanoic acid (11-MUA). Apart from the method of aptamer immobilization, the biosensors differ in the signal generation. To date, most extensively studied were the biosensors based on optical methods (fluorescence, SPR) and acoustic sensors based mostly on thickness shear mode (TSM) method. However, recently several investigators reported electrochemical sensors based on enzyme-labeled aptamers, electrochemical indicators and impedance spectroscopy methods of detection. 

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