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Registration Number

0224U032660, (0123U102859) , R & D reports

Title

Development of an enzyme kit and a portable biosensor for express assay of creatinine – a marker of acute renal dysfunction

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Скринінг та синтез нових хемосенсорних матеріалів, чутливих до іонів амонію. Розробка оптимальних композицій КДІ та наночасток для покращення чутливості та стабільності ензиматичних систем. Розробка та характеризація нових амперометричного та потенціометричного біосенсорів для визначення вмісту креатиніну на основі використання рекомбінантної КДІ.

Head

Honchar Mykhailo V., д.б.н.

Registration Date

29-11-2024

Organization

Institute of Cell Biology of the National Academy of Sciences of Ukraine

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The aim of the project is the synthesis, study of structural, physicochemical and catalytic properties of new catalytically active nanomaterials, namely, artificial enzymes (nanozymes); creation on their basis of new diagnostic methods and study of biological action of nanocomposites on eukaryotic cells (in particular, yeast). The creation of new nanomaterials with catalytic properties will allow the development of new methods of highly sensitive quantitative detection of a number of practically important substances (glucose, lactic acid, alcohols) for clinical diagnosis, sports medicine and food technology. The relevance of the planned research is related to the need in development of selective, sensitive and at the same time inexpensive methods for analysis of practically important analytes - biomarkers of the most common diseases or indicators of food quality. Socioeconomic significance of the planned work is the need to replace natural enzymes with inexpensive artificial catalysts in bio/chemosensors and analytical kits, as well as improving the sensitivity, assay time and cost of sensor systems by including nanoparticles with catalytic activity (nanozymes) in the recognition layer.

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 The object of the research – creatinine deiminase and creatinine. Subject of the research – biosensors and enzymatic approaches for determination of creatinine. The purpose of the work - the development of new bioanalytical methods for quantitative determination of creatinine - an important biomarker of the functional state of the kidneys. Research methods: chemical, physicochemical, biochemical, microbiological, enzymatic, scanning electron microscopy, energy-dispersive X-ray spectroscopy, UV-visible spectroscopy, X-Ray photoelectron spectroscopy (XPS), cyclic voltammetry, chronoamperometry, methods of synthesis of nanomaterials, enzyme immobilization methods, statistical analysis. Results and novelty: For the first time: – an amperometric bionanosensor for analysis of creatinine based on recombinant creatinine deiminase and nanochelator nanoparticles nCoCu, sensitive to N-methylhydantoin was developed; – the structure of the redox-active nCoCu-N-methylhydantoin complex was identified for the first time; – for the first time, a new strategy for additional enrichment of recombinant bacterial cells Corynobacterium glutamicum with a target enzyme co-immobilized with nanoparticles nCu, was applied for the development of a Crn-selective whole cell sensor; Improved: – methodology for obtaining a new metal-hybrid nanomaterials sensitive to ammonium ions and N-methylhydantoin; – method of immobilization of creatinine deiminase and bacterial cells on the surface of electrochemical converters; – method of construction of conductometric bisensors based on ion-sensitive field effect transistors. The product of this stage of work – scientific publications. There was published 14 articles in international (8) and domestic journals (6) with a total impact factor of 44.248. An application for a utility model and an application for an invention have been submitted.

Product Description

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Demkiv Olha M.
Zakalska Oksana M.
Zakalskyi Andrii Ye.
Serkiz Roman Ya.
Stasiuk Nataliia Ye.

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2024-11-29