期刊
ANALYTICAL BIOCHEMISTRY
卷 385, 期 2, 页码 334-341出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2008.10.037
关键词
Glucose biosensor; Microband electrode; Cell culture; HepG2 cells; Glucose metabolism
资金
- Research Councils UK (RCUK, Basic Technology Grant) [EP/C52389X/1]
Microband glucose biosensors were produced by insulating and sectioning through a screen-printed, water-based carbon electrode containing cobalt phthalocyanine redox mediator and glucose oxidase enzyme. Under quiescent conditions at 37 degrees C, at an operating potential of +0.4 V, they produced an amperometric response to glucose in buffer solutions with a sensitivity of 26.4 nA/mM and a linear range of 0.45 to 9.0 mM. An optimal pH value of 8.5 was obtained under these conditions, and a value for activation energy of 40.55 kJ mol(-1) was calculated. In culture medium (pH 7.3), a sensitivity of 13 nA/mM was obtained and the response was linear up to 5 mM with a detection limit of 0.5 mM. The working concentration was up to 20 mM glucose with a precision of 11.3% for replicate biosensors (n = 4). The Microband biosensors were applied to determine end-point glucose concentrations in Culture medium by monitoring steady-state current responses 400 s after transfer of the biosensors into different sample solutions. In conjunction with Cultures of HepG2 (human Caucasian hepatocyte carcinoma) cells, current responses obtained in 24-h supernatants showed an inverse correlation (R-2 = 0.98) with cell number, indicating that the biosensors were applicable for monitoring glucose metabolism by cells and of quantifying cell number. Glucose concentrations determined using the biosensor assay were in good agreement, for concentrations Lip to 20 mM, with those determined spectrophotometrically (R-2 = 0.99). This method of end-point glucose determination was used to provide an estimated rate of glucose uptake for HepG2 cells of 7.9 nmol/(10(6) cells min) based on a 24-h period in Culture. (C) 2008 Elsevier Inc. All rights reserved.
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