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Registration Number

0225U004272, (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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Конструювання лабораторних прототипів амперометричного та потенціометричного біосенсорів для експрес- аналізу креатиніну, порівняння їхніх аналітичних характеристик. Дослідження феномену деградації та надійності ензимних біосенсорів при різних умовах аналізу та зберігання. Тестування створених біотехнологічних продуктів – ензиматичного набору та біосенсорів для аналізу креатиніну – на реальних клінічних зразках біологічних рідин людини. Валідація отриманих результатів за допомогою традиційних методів.

Head

Honchar Mykhailo V., д.б.н.

Registration Date

27-11-2025

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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An enzymatic method and amperometric bionanosensors for the analysis of creatinine content using recombinant creatine deiminase (CDI) and N-methylhydantoin and ammonium chelators – nCoCu, nCu, nCu-ZIF have been developed. The structure of the redox-active complex nCoCu-N-methylhydantoin has been identified for the first time. A new strategy for creating cell-based sensors using genetically modified bacterium C. glutamicum enriched with the target enzyme and nanomaterials has been proposed. An amperometric biosensor for the simultaneous analysis of creatinine and sarcosine content using co-immobilized preparations of CDI, sarcosine oxidase (SOx) and nCu-ZIF nanoparticles sensitive to N-methylhydantoin and hydrogen peroxide has been developed for the first time. The designed biosensor operates at a potential of +50 mV (for the analysis of creatinine) and at –200 mV (for sarcosine). The developed biosensors and enzymatic method based on purified preparations of the enzyme KDI and SOx, as well as o-phenylenediamine as a chromogen, were used for the quantitative analysis of creatinine in real samples, namely, for the analysis of creatinine and sarcosine in clinical samples of biological fluids of healthy donors and patients with various nephrological diseases: pyelonephritis, glomerulonephritis and diabetic nephropathy. The possibility of using the created bioanalytical methods for non-invasive assay of creatinine in human sweat, saliva and urine was also studied. The results of determining the creatinine content in human biological samples by the developed methods were compared with the results of analysis of the same samples by reference approaches. Published 20 articles (11 in international and 9 in domestic journals), including 11 papers in journals classified in the first and second quartiles (Q1 and Q2) .

Product Description

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Roman Y. Serkiz
Olha M. Demkiv
Oksana M. Zakalska
Andriy Y. Zakalskiy
Nataliya Y. Stasyuk
Mykailo V. Gonchar

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2025-11-27