Performance comparison between multienzymes loaded single and dual electrodes for the simultaneous electrochemical detection of adenosine and metabolites in cancerous cells
| Autor: | Dong-Keun Jung, Khalil K. Hussain, Yoon-Bo Shim, Moo Hyun Kim, Mahmood H. Akhtar |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Xanthine Oxidase
Adenosine Immobilized enzyme Adenosine Deaminase Biomedical Engineering Biophysics Purine nucleoside phosphorylase Biosensing Techniques 02 engineering and technology 01 natural sciences Michaelis–Menten kinetics chemistry.chemical_compound Adenosine deaminase Limit of Detection Neoplasms Electrochemistry medicine Humans Metabolomics Inosine Electrodes Hypoxanthine Detection limit biology Chemistry 010401 analytical chemistry General Medicine 021001 nanoscience & nanotechnology 0104 chemical sciences Purine-Nucleoside Phosphorylase A549 Cells biology.protein 0210 nano-technology Biotechnology medicine.drug Nuclear chemistry |
| Zdroj: | Biosensors and Bioelectronics. 109:263-271 |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/j.bios.2018.03.031 |
| Popis: | The analytical performance of the multi enzymes loaded single electrode sensor (SES) and dual electrode sensor (DES) was compared for the detection of adenosine and metabolites. The SES was fabricated by covalent binding of tri-enzymes, adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP), and xanthine oxidase (XO) along with hydrazine (Hyd) onto a functionalized conducting polymer [2,2:5,2-terthiophene-3-(p-benzoic acid)] (pTTBA). The enzyme reaction electrode in DES was fabricated by covalent binding of ADA and PNP onto pTTBA coated on Au nanoparticles. The detection electrode in DES was constructed by covalent binding of XO and Hyd onto pTTBA coated on porous Au. Due to the higher amount (3.5 folds) of the immobilized enzymes and Hyd onto the DES than SES, and the lower Michaelis constant (Km) value for DES (28.7 µM) compared to SES (36.1 µM), the sensitivity was significantly enhanced for the DES (8.2 folds). The dynamic range obtained using DES was from 0.5 nM to 120.0 µM with a detection limit of 1.43 nM ± 0.02, 0.76 nM ± 0.02, and 0.48 nM ± 0.01, for adenosine (AD), inosine (IN), and hypoxanthine (Hypo) respectively. Further, the DES was coupled with an electrochemical potential modulated microchannel for the separation and simultaneous detection of AD, IN, and Hypo in an extracellular matrix of cancerous (A549) and non-cancerous (Vero) cells. The sensor probe confirms a higher basal level of extracellular AD and its metabolites in cancer cells compared to normal cells. In addition, the effect of dipyridamole on released adenosine in A549 cells was investigated. |
| Databáze: | OpenAIRE |
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