Abstract
In the work, the possibility of using nanoparticles of gold (AuNPs) to upgrade bioselective elements of biosensors in order to improve their analytical characteristics is considered. The bioselective elements of biosensors based on acetylcholinesterase (AChE), butyryl cholinesterase (BuChE) and glucose oxidase (GOD) were used as an experimental model. Immobilization of enzymes on the surfaces of conductometric transducers was performed by the crosslinking of corresponding enzymes using glutaraldehyde. The conditions of immobilization of AChE with gold nanoparticles were optimized. Thus, we determined the optimal values of concentration of crosslinking agent (glutaraldehyde), duration of immobilization, the enzyme to AuNPs ratio, the AuNPs concentration and size. The performance characteristics of the biosensors based on enzymes and AuNPs were investigated and compared with the characteristics of biosensors based on enzymes only. It was also examined how the addition of AuNPs to the bioselective element of biosensors affects the biosensor stability. In particular, the reproducibility of preparation and continuous operation of biosensors was tested as well as their stability at storage. It was shown that the presence of AuNPs in the composition of bioselective elements can improve some characteristics of biosensors, which may be promising for further study and use.
Highlights
The development of new methods for diagnosing diseases of humans and farm animals is an extremely urgent task of biotechnology
At the first stage of this work, it was necessary to optimize the conditions of immobilization of AChE with nanoparticles that will allow consequent comparison of the analytical characteristics of the obtained biosensor with those of the biosensor based on the traditional method of immobilization
The immobilization of AChE with gold nanoparticles on the surface of conductometric transducers was performed in a solution of glutaraldehyde; the latter formed covalent bonds between the enzyme and auxiliary substance (BSA); during this process, the enzyme activity changed, depending on the GA concentration and the duration of immobilization
Summary
The development of new methods for diagnosing diseases of humans and farm animals is an extremely urgent task of biotechnology. A promising approach is the use of biosensors, new analytical devices combining the achievements in biology, physics, chemistry, mathematics, microelectronics, etc. One of the options to improve the analytical characteristics. 2 F.D. Ovcharenko Institute of Biocolloidal Chemistry, NAS of Ukraine, Vernadskogo Ave, 42, Kyiv 03142, Ukraine. Gold nanoparticles (AuNPs) can be widely used in enzyme-based biosensors. AuNPs are highly conductive and biocompatible, they are able to form strong thiol bonds with organic substances (eg, cysteine residues of enzymes) (Kerman et al 2008). Nanoparticles form a suitable microenvironment for enzyme immobilization. The activity of the immobilized enzyme can be preserved by immobilization onto AuNPs (Hondred et al 2017)
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