Development and Optimisation of the Biosensor for Aspartate Aminotransferase Blood Level Determination

Overview

Journal Anal Bioanal Chem

Specialty Chemistry

Date 2024 Dec 12

PMID 39666000

Authors

Daryna Mruga

Sergei Dzyadevych

Oleksandr Soldatkin

Affiliations

Soon will be listed here.

Abstract

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.

References

Thupakula S, Nimmala S, Ravula H, Chekuri S, Padiya R . Emerging biomarkers for the detection of cardiovascular diseases. Egypt Heart J. 2022; 74(1):77. PMC: 9584006. DOI: 10.1186/s43044-022-00317-2. View

Mera J, Dickson B, Feldman M . Influence of gender on the ratio of serum aspartate aminotransferase (AST) to alanine aminotransferase (ALT) in patients with and without hyperbilirubinemia. Dig Dis Sci. 2007; 53(3):799-802. DOI: 10.1007/s10620-007-9924-z. View

Liu X, Liu P . Elevated AST/ALT ratio is associated with all-cause mortality in patients with stable coronary artery disease: a secondary analysis based on a retrospective cohort study. Sci Rep. 2022; 12(1):9231. PMC: 9163143. DOI: 10.1038/s41598-022-13355-2. View

Xuan G, Oh S, Choi E . Development of an electrochemical immunosensor for alanine aminotransferase. Biosens Bioelectron. 2003; 19(4):365-71. DOI: 10.1016/s0956-5663(03)00194-5. View

Sha H, Yan B . Dye-functionalized metal-organic frameworks with the uniform dispersion of MnO nanosheets for visualized fluorescence detection of alanine aminotransferase. Nanoscale. 2021; 13(47):20205-20212. DOI: 10.1039/d1nr05376k. View

Resmi P, Sachin Kumar S, Alageswari D, Suneesh P, Ramachandran T, Nair B . Development of a paper-based analytical device for the colourimetric detection of alanine transaminase and the application of deep learning for image analysis. Anal Chim Acta. 2021; 1188:339158. DOI: 10.1016/j.aca.2021.339158. View

Hsueh C, Wang J, Dai L, Liu C . Development of an electrochemical-based aspartate aminotransferase nanoparticle ir-C biosensor for screening of liver diseases. Biosensors (Basel). 2015; 2(2):234-44. PMC: 4263573. DOI: 10.3390/bios2020234. View

Chang K, Chang C, Chou S, Chen C . Sequential measurement of aminotransferase activities by amperometric biosensors. Biosens Bioelectron. 2007; 22(12):2914-20. DOI: 10.1016/j.bios.2006.12.001. View

Song M, Yun D, Min N, Hong S . Electrochemical biosensor array for liver diagnosis using silanization technique on nanoporous silicon electrode. J Biosci Bioeng. 2007; 103(1):32-7. DOI: 10.1263/jbb.103.32. View

10.

Wang H, Chu C, Tsai S, Yang R . Aspartate Aminotransferase and Alanine Aminotransferase Detection on Paper-Based Analytical Devices with Inkjet Printer-Sprayed Reagents. Micromachines (Basel). 2018; 7(1). PMC: 6189706. DOI: 10.3390/mi7010009. View

11.

Zhang M, Karra S, Gorski W . Electrochemical coupled-enzyme assays at carbon nanotubes. Anal Chem. 2014; 86(18):9330-4. DOI: 10.1021/ac502687z. View

12.

Kucherenko I, Didukh D, Soldatkin O, Soldatkin A . Amperometric biosensor system for simultaneous determination of adenosine-5'-triphosphate and glucose. Anal Chem. 2014; 86(11):5455-62. DOI: 10.1021/ac5006553. View

13.

Saito K, Sugawara H, Watanabe T, Ishii A, Fukuchi T . A retrospective cross-sectional study for predicting 72-h mortality in patients with serum aspartate aminotransferase levels ≥ 3000 U/L. Sci Rep. 2021; 11(1):800. PMC: 7804105. DOI: 10.1038/s41598-020-79435-3. View

14.

Han Y, Song S, Lee J, Lee D, Yoon H . Multienzyme-modified biosensing surface for the electrochemical analysis of aspartate transaminase and alanine transaminase in human plasma. Anal Bioanal Chem. 2011; 400(3):797-805. DOI: 10.1007/s00216-011-4797-6. View

15.

Sang J, Cheng J, Hu H, Liu K, Guo J, Guo J . Portable dual-channel blood enzyme analyzer for point-of-care liver function detection. Analyst. 2023; 148(23):6020-6027. DOI: 10.1039/d3an01432k. View

16.

Saygili E, Orakci B, Koprulu M, Demirhan A, Ilhan-Ayisigi E, Kilic Y . Quantitative determination of HO for detection of alanine aminotransferase using thin film electrodes. Anal Biochem. 2019; 591:113538. DOI: 10.1016/j.ab.2019.113538. View