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Information × Registration Number 0219U001826, 0116U002208 , R & D reports Title Development of new biosensor and enzymatic methods for determination of the content of creatinine as a biomarker of kidney function and control process of hemodialysis popup.stage_title Head Gonchar Mykhailo Vasyl'ovych, Кандидат педагогічних наук Registration Date 06-03-2019 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 The object of research: recombinant enzymes of bioanalytical significance, biosensor analysis, enzymatic analysis, creatinine, biological samples. The aim of investigations: optimization of biosensor and enzymatic methods of creatinine analysis on real clinical samples of biological fluids for assessing the functional state of the kidneys and the effectiveness of hemodialysis.Methods: biochemical, chromatographic, bioelectrochemical and enzimological methods, approaches of protein chemistry and analytical biotechnology. The results of the work: By the use of CDI and spectrofluorimetric method for monitoring the product of the enzymatic reaction (ammonium ions), optimization of simultaneous analysis of creatinine and ammonia in human blood serum was performed. The optimization parameters were concentrations of CDI, o-phthaldehyde, EDTA, the type of buffer system, the method of preparation of samples for analysis. An amperometric biosensor for quantitative determination of creatinine was constructed on the basis of CDI and Zn(Hg)S/Cu nanoparticles. The created biosensor is characterized by a high selectivity and sensitivity to creatinine (90.8 ± 2.8 A ? M-1 ? m-2). The linear range of the biosensor is observed in the range from 0.03 mM to 3.5 mM with a low limit of determination for creatinine 20 ?M. Another biosensor with a higher sensitivity to creatinine (424 ± 20 A ? M-1 ? m-2) was constructed by the use of an alternative ammonium-sensitive chemosensory membrane - electrodeposited polyaniline (PAN) in a complex with Nafion. The linear range of the biosensor is observed from 0.015 mM to 0.9 mM with a low limit of determination for creatinine 12 ?M. The developed biosensors, based on recombinant CDI, have been successfully tested for simultaneous analysis of creatinine and ammonium in real samples of blood serum. A comparative analysis of the results of creatinine determination in serum samples of patients with impaired renal function before and after the hemodialysis procedure by the use of biosensor and enzymatic-fluorimetric methods showed a high level of correlation of analytical data (R = 0.998), which evidences the reliability and accuracy of the proposed approaches. A laboratory prototype of the bioanalytical kit for enzymatic-fluorimetric analysis of creatinine and ammonia in biological human fluids was created, the composition of the components was optimized and the Manual for the use of the kit was proposed. Product Description popup.authors Іваш М. Ф. Гайда Г. З. Демків О.М. Закальська О.М. Закальський А.Є. Куцяба В.І. Прокопів Т.М. Серкіз Р.Я. Смуток О.В. Стасюк Н. Є. Стасюк Н.Є. popup.nrat_date 2020-04-02 Close