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Performance comparison between multienzymes loaded single and dual electrodes for the simultaneous electrochemical detection of adenosine and metabolites in cancerous cells
Biosensors and Bioelectronics ( IF 12.6 ) Pub Date : 2018-03-15 , DOI: 10.1016/j.bios.2018.03.031
Khalil K. Hussain , Mahmood H. Akhtar , Moo-Hyun Kim , Dong-Keun Jung , Yoon-Bo Shim

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.



中文翻译:

负载多酶的单电极和双电极之间的性能比较,用于同时电化学检测癌细胞中的腺苷和代谢产物

比较了多酶负载的单电极传感器(SES)和双电极传感器(DES)的分析性能,以检测腺苷和代谢产物。通过将三酶,腺苷脱氨酶(ADA),嘌呤核苷磷酸化酶(PNP)和黄嘌呤氧化酶(XO)与肼(Hyd)共价结合到功能化的导电聚合物上来制备SES [2,2:5,2 -对噻吩-3-(对苯甲酸)](pTTBA)。通过将ADA和PNP共价结合到包被在Au纳米颗粒上的pTTBA上,制得DES中的酶反应电极。通过XO和Hyd共价结合到涂在多孔Au上的pTTBA上来构建DES中的检测电极。由于与SES相比,固定在DES上的酶和Hyd含量更高(3.5倍),并且DES的米氏常数(Km)值较低(28。与SES(36.1 µM)相比(7 µM),DES的灵敏度显着提高(8.2倍)。对于腺苷(AD),肌苷(IN)和次黄嘌呤(Hypo),使用DES获得的动态范围为0.5 nM至120.0 µM,检出限为1.43 nM±0.02、0.76 nM±0.02和0.48 nM±0.01。分别。此外,DES与电化学势调制微通道耦合,用于分离和同时检测癌细胞(A549)和非癌细胞(Vero)的细胞外基质中的AD,IN和Hypo。传感器探针证实,与正常细胞相比,癌细胞中细胞外AD及其代谢物的基础水平更高。另外,研究了双嘧达莫对A549细胞中释放的腺苷的影响。对于腺苷(AD),肌苷(IN)和次黄嘌呤(Hypo),使用DES获得的动态范围为0.5 nM至120.0 µM,检出限为1.43 nM±0.02、0.76 nM±0.02和0.48 nM±0.01。分别。此外,DES与电化学势调制微通道耦合,用于分离和同时检测癌细胞(A549)和非癌细胞(Vero)的细胞外基质中的AD,IN和Hypo。传感器探针证实,与正常细胞相比,癌细胞中细胞外AD及其代谢物的基础水平更高。另外,研究了双嘧达莫对A549细胞中释放的腺苷的影响。对于腺苷(AD),肌苷(IN)和次黄嘌呤(Hypo),使用DES获得的动态范围为0.5 nM至120.0 µM,检出限为1.43 nM±0.02、0.76 nM±0.02和0.48 nM±0.01。分别。此外,DES与电化学势调制微通道耦合,用于分离和同时检测癌细胞(A549)和非癌细胞(Vero)的细胞外基质中的AD,IN和Hypo。传感器探针证实,与正常细胞相比,癌细胞中细胞外AD及其代谢物的基础水平更高。另外,研究了双嘧达莫对A549细胞中释放的腺苷的影响。和次黄嘌呤(Hypo)。此外,DES与电化学势调制微通道耦合,用于分离和同时检测癌细胞(A549)和非癌细胞(Vero)的细胞外基质中的AD,IN和Hypo。传感器探针证实,与正常细胞相比,癌细胞中细胞外AD及其代谢物的基础水平更高。另外,研究了双嘧达莫对A549细胞中释放的腺苷的影响。和次黄嘌呤(Hypo)。此外,DES与电化学势调制微通道耦合,用于分离和同时检测癌细胞(A549)和非癌细胞(Vero)的细胞外基质中的AD,IN和Hypo。传感器探针证实,与正常细胞相比,癌细胞中细胞外AD及其代谢物的基础水平更高。此外,研究了双嘧达莫对A549细胞中腺苷释放的影响。传感器探针证实,与正常细胞相比,癌细胞中细胞外AD及其代谢物的基础水平更高。另外,研究了双嘧达莫对A549细胞中释放的腺苷的影响。传感器探针证实,与正常细胞相比,癌细胞中细胞外AD及其代谢物的基础水平更高。另外,研究了双嘧达莫对A549细胞中释放的腺苷的影响。

更新日期:2018-03-15
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