Mruga Daryna, Dzyadevych Sergei, Soldatkin Oleksandr
Institute of Molecular Biology and Genetics of the National Academy of Sciences of Ukraine, Zabolotnogo Street 150, Kyiv, 03680, Ukraine.
Taras Shevchenko National University of Kyiv, Volodymyrska Street 64, Kyiv, 01003, Ukraine.
Anal Bioanal Chem. 2025 Feb;417(4):721-731. doi: 10.1007/s00216-024-05682-2. Epub 2024 Dec 12.
This work presents the development and optimisation of an amperometric biosensor for determining aspartate aminotransferase (AST) activity in blood serum, using glutamate oxidase and platinum disc electrodes. AST is a key biomarker for diagnosing cardiovascular and liver diseases. The biosensor's bioselective membrane composition and formation protocol and the working solution (aspartate 8 mM, α-ketoglutarate 2 mM, pyridoxal-5-phosphate 100 µM) were optimised. The sensor demonstrated high selectivity, stability (70% retention over 2 months at - 18 °C), and sensitivity (2.37 nA min⁻ per 10 U L⁻), with a dynamic range of 0-500 U L⁻ and a limit of detection of 1 U L⁻. Comparative analysis showed the calibration curve method outperforms the standard addition method for AST measurement in serum samples. Additionally, a reference spectrophotometric technique was adapted for AST level determination, showing a strong correlation (r = 0.989) with the biosensor results. This research offers a fast, affordable, and accurate tool for early check-ups of liver and heart conditions. The biosensor's flexibility and ease of use make it suitable for further development into point-of-care testing and personalised healthcare techniques.
这项工作展示了一种用于测定血清中天冬氨酸转氨酶（AST）活性的电流型生物传感器的开发与优化，该传感器使用了谷氨酸氧化酶和铂盘电极。AST是诊断心血管疾病和肝脏疾病的关键生物标志物。对生物传感器的生物选择性膜组成、形成方案以及工作溶液（天冬氨酸8 mM、α-酮戊二酸2 mM、磷酸吡哆醛100 µM）进行了优化。该传感器具有高选择性、稳定性（在-18°C下2个月内保留70%）和灵敏度（每10 U L⁻为2.37 nA min⁻），动态范围为0 - 500 U L⁻，检测限为1 U L⁻。对比分析表明，在血清样本中AST测量方面，校准曲线法优于标准加入法。此外，采用了一种参考分光光度技术来测定AST水平，其与生物传感器结果显示出很强的相关性（r = 0.989）。本研究为肝脏和心脏状况的早期检查提供了一种快速、经济且准确的工具。该生物传感器的灵活性和易用性使其适合进一步发展为即时检测和个性化医疗技术。
