一种用于检测多巴胺的新型辣根过氧化物酶生物传感器：伏安法研究。
A novel horseradish peroxidase biosensor towards the detection of dopamine: a voltammetric study.
作者信息
Raghu P, Reddy T Madhusudana, Gopal P, Reddaiah K, Sreedhar N Y
机构信息
Electrochemical Research Laboratory, Department of Chemistry, S.V.U. College of Sciences, Sri Venkateswara University, Tirupati, 517502, Andhra Pradesh, India.
Electrochemical Research Laboratory, Department of Chemistry, S.V.U. College of Sciences, Sri Venkateswara University, Tirupati, 517502, Andhra Pradesh, India.
出版信息
Enzyme Microb Technol. 2014 Apr 10;57:8-15. doi: 10.1016/j.enzmictec.2014.01.002. Epub 2014 Jan 13.
A polymerized film of glycine (Gly) was prepared on the surface of carbon paste electrode (CPE) through the cyclic voltammetry (CV) technique. A novel biosensor for the determination of dopamine (DA) has been constructed based on horseradish peroxidase (HRP) and multiwalled carbon nanotubes (MWCNTs) immobilizing on Poly (Gly)/CPE through silica sol-gel (SiSG) entrapment. CV measurements were employed in order to understand the feasibility of poly (Gly) as an electron carrier between the immobilized peroxidase and the surface of CPE. By using differential pulse voltammetry (DPV) the calibration curves of DA was obtained in the range of 15-865 μM. The limit of detection (LOD) and limit of quantification (LOQ) of DA was found to be 6×10⁻⁷ M and 2×10⁻⁶ M respectively. The apparent Michaelis-Menten constant (Km(app)) was found to be 0.5 mM and illustrated that the good biological activity of the fixed enzyme. Electrochemical impedance spectroscopy (EIS) results confirmed the rapid electron transfer and also the immobilization of enzyme on the electrode surface. The biosensor showed high sensitivity, selectivity and reproducibility. This method has been used to determine DA in the presence of various interferences and in clinical preparations.
通过循环伏安法（CV）技术在碳糊电极（CPE）表面制备了甘氨酸（Gly）的聚合膜。基于辣根过氧化物酶（HRP）和通过硅溶胶 - 凝胶（SiSG）包埋固定在聚（Gly）/ CPE上的多壁碳纳米管（MWCNTs）构建了一种用于测定多巴胺（DA）的新型生物传感器。采用循环伏安法测量以了解聚（Gly）作为固定化过氧化物酶与CPE表面之间电子载体的可行性。通过差分脉冲伏安法（DPV）获得了在15 - 865μM范围内的DA校准曲线。发现DA的检测限（LOD）和定量限（LOQ）分别为6×10⁻⁷ M和2×10⁻⁶ M。表观米氏常数（Km(app)）为0.5 mM，表明固定化酶具有良好的生物活性。电化学阻抗谱（EIS）结果证实了快速的电子转移以及酶在电极表面的固定化。该生物传感器具有高灵敏度、选择性和重现性。该方法已用于在存在各种干扰的情况下以及临床制剂中测定DA。
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