Endocrinology and Metabolism Research Center, Tehran University of Medical Sciences, P.O. Box 14395/1179, Tehran, Islamic Republic of Iran.
Mol Biol Rep. 2013 Mar;40(3):2327-34. doi: 10.1007/s11033-012-2314-4. Epub 2012 Nov 28.
3-Hydroxybutyrate, one of the main blood ketone bodies, has been considered as a critical indicator for diagnosis of diabetic ketoacidosis. Biosensors designed for detection of 3-hydroxybutyrate with advantages of precision, easiness and speedy performance have attracted increasing attention. This study attempted to develop a 3-hydroxybutyrate dehydrogenase-based biosensor in which single-walled carbon nanotubes (SWCNT) was used in order to immobilize the cofactor, NAD(+), on the surface of screen-printed electrode. The formation of NAD(+)-SWCNT conjugates was assessed by electrochemistry and electron microscopy. Cyclic voltammetry was used to analyze the performance of this biosensor electrochemically. The considerable shelf life and reliability of the proposed biosensor to analyze real sample was confirmed by this method. The reduction in the over potential of electrochemical oxidation of NADH to -0.15 V can be mentioned as a prominent feature of this biosensor. This biosensor can detect 3-hydroxybutyrate in the linear range of 0.01-0.1 mM with the low detection limit of 0.009 mM. Simultaneous application of screen-printed electrode and SWCNT has made the biosensor distinguished which can open new prospects for detection of other clinically significant metabolites.
3-羟基丁酸是主要的血液酮体之一,已被认为是诊断糖尿病酮症酸中毒的一个关键指标。具有精度高、操作简单、检测速度快等优点的 3-羟基丁酸生物传感器受到了越来越多的关注。本研究试图开发一种基于 3-羟丁酸脱氢酶的生物传感器,其中单壁碳纳米管(SWCNT)用于将辅因子 NAD(+)固定在丝网印刷电极的表面。通过电化学和电子显微镜评估了 NAD(+)-SWCNT 缀合物的形成。循环伏安法用于分析该生物传感器的电化学性能。通过该方法证实了该生物传感器在分析实际样品时具有相当长的保质期和可靠性。电化学氧化 NADH 的过电势降低至-0.15 V 可以说是该生物传感器的一个显著特点。该生物传感器可以在 0.01-0.1 mM 的线性范围内检测 3-羟基丁酸,检测限低至 0.009 mM。丝网印刷电极和 SWCNT 的同时应用使该生物传感器具有区分能力,这为检测其他临床上有意义的代谢物开辟了新的前景。