Arya Sunil K, Prusty Arun K, Singh S P, Solanki Pratima R, Pandey Manoj K, Datta Monika, Malhotra Bansi D
Biomolecular Electronics and Conducting Polymer Research Group, National Physical Laboratory, K. S. Krishnan Marg, New Delhi 110012, India; Department of Chemistry, University of Delhi, Delhi 110007, India.
Anal Biochem. 2007 Apr 15;363(2):210-8. doi: 10.1016/j.ab.2007.01.029. Epub 2007 Jan 26.
Cholesterol oxidase (ChOx) has been covalently immobilized onto two-dimensional self-assembled monolayer (SAM) of N-(2-aminoethyl)-3-aminopropyl-trimethoxysilane (AEAPTS) deposited on the indium-tin oxide (ITO) coated glass plates using N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide and N-hydroxysuccinimide (EDC/NHS) chemistry. These ChO x/AEAPTS/ITO bioelectrodes are characterized using contact angle (CA) measurements, UV-visible spectroscopy, atomic force microscopy (AFM), electrochemical impedance technique, and Fourier transform infrared (FT-IR) technique. The covalently immobilized ChOx-modified AEAPTS bioelectrodes are used for the estimation of cholesterol in solution using UV-visible technique. These cholesterol sensing bioelectrodes show linearity as 50 to 500 mg/dl for cholesterol solution, detection limit as 25mg/dl, sensitivity as 4.499 x 10(-5) Abs (mg/dl)(-1), K(m) value as 58.137 mg/dl (1.5mM), apparent enzyme activity as 1.81 x 10(-3) U cm(-2), shelf life of approximately 10 weeks, and electrode reusability as 10 times.
胆固醇氧化酶（ChOx）已通过N-乙基-N'-(3-二甲氨基丙基)碳二亚胺和N-羟基琥珀酰亚胺（EDC/NHS）化学方法共价固定在沉积于氧化铟锡（ITO）涂层玻璃板上的N-(2-氨基乙基)-3-氨丙基三甲氧基硅烷（AEAPTS）二维自组装单分子层（SAM）上。这些ChOx/AEAPTS/ITO生物电极通过接触角（CA）测量、紫外可见光谱、原子力显微镜（AFM）、电化学阻抗技术和傅里叶变换红外（FT-IR）技术进行表征。共价固定的ChOx修饰的AEAPTS生物电极用于通过紫外可见技术测定溶液中的胆固醇。这些胆固醇传感生物电极对胆固醇溶液的线性范围为50至500mg/dl，检测限为25mg/dl，灵敏度为4.499×10⁻⁵ Abs (mg/dl)⁻¹，米氏常数（Kₘ）值为58.137mg/dl（1.5mM），表观酶活性为1.81×10⁻³ U cm⁻²，保质期约为10周，电极可重复使用10次。
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