Wang Qianyu, Liu Yujie, Campillo-Brocal Jonatan C, Jiménez-Quero Amparo, Crespo Gaston A, Cuartero María
Department of Chemistry, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, Teknikringen 30, SE-100 44, Stockholm, Sweden.
Department of Genetics and Microbiology, University of Murcia, Campus Universitario de Espinardo, Murcia, Spain.
Biosens Bioelectron. 2021 Jun 15;182:113154. doi: 10.1016/j.bios.2021.113154. Epub 2021 Mar 11.
We present herein the very first amperometric biosensor for the quantitative determination of glycine in diverse biological fluids. The biosensor is based on a novel quinoprotein that catalyzes the oxidation of glycine with high specificity. This process is coupled to the redox conversion of Prussian blue in the presence of hydrogen peroxide originating from the enzymatic reaction. The optimized tailoring of the biosensor design consists of the effective encapsulation of the quinoprotein in a chitosan matrix with the posterior addition of an outer Nafion layer, which is here demonstrated to suppress matrix interference. This is particularly important in the case of ascorbic acid, which is known to influence the redox behavior of the Prussian blue. The analytical performance of the biosensor demonstrates fast response time (<7 s), acceptable reversibility, reproducibility, and stability (<6% variation) as well as a wide linear range of response (25-500 μM) that covers healthy (and even most unhealthy) physiological levels of glycine in blood/serum, urine and sweat. A total of 6 real samples from healthy patients and animals were analyzed: two serum, two urine and two sweat samples. The results were validated via commercially available fluorescence kit, displaying discrepancy of less than 9% in all the samples. The unique analytical features and effortless preparation of the new glycine biosensor position it at the forefront of current technologies towards decentralized clinical applications and sport performance monitoring.
我们在此展示了首个用于定量测定多种生物体液中甘氨酸的电流型生物传感器。该生物传感器基于一种新型醌蛋白，它能高特异性地催化甘氨酸的氧化。在源于酶促反应产生的过氧化氢存在下，这一过程与普鲁士蓝的氧化还原转化相偶联。生物传感器设计的优化剪裁包括将醌蛋白有效封装在壳聚糖基质中，并随后添加外部的Nafion层，在此已证明这可抑制基质干扰。这在抗坏血酸的情况下尤为重要，已知抗坏血酸会影响普鲁士蓝的氧化还原行为。该生物传感器的分析性能显示出快速响应时间（<7秒）、可接受的可逆性、重现性和稳定性（<6%变化）以及宽线性响应范围（25 - 500μM），涵盖了血液/血清、尿液和汗液中甘氨酸的健康（甚至大多数不健康）生理水平。共分析了来自健康患者和动物的6个真实样本：两个血清样本、两个尿液样本和两个汗液样本。结果通过市售荧光试剂盒进行了验证，所有样本中的差异均小于9%。新型甘氨酸生物传感器独特的分析特性和简便的制备方法使其处于当前分散式临床应用和运动表现监测技术的前沿。
