Mikuła Edyta, Wysłouch-Cieszyńska Aleksandra, Zhukova Liliya, Puchalska Monika, Verwilst Peter, Dehaen Wim, Radecki Jerzy, Radecka Hanna
Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland.
Institute of Biochemistry and Biophysics of Polish Academy of Sciences, Pawińskiego 5a, 02-106 Warsaw, Poland.
Sensors (Basel). 2014 Jun 17;14(6):10650-63. doi: 10.3390/s140610650.
In this work we report on an electrochemical biosensor for the determination of the S100B protein. The His-tagged VC1 domains of Receptors for Advanced Glycation End (RAGE) products used as analytically active molecules were covalently immobilized on a monolayer of a thiol derivative of pentetic acid (DPTA) complex with Cu(II) deposited on a gold electrode surface. The recognition processes between the RAGE VC1 domain and the S100B protein results in changes in the redox activity of the DPTA-Cu(II) centres which were measured by Osteryoung square-wave voltammetry (OSWV). In order to verify whether the observed analytical signal originates from the recognition process between the His6-RAGE VC1 domains and the S100B protein, the electrode modified with the His6-RAGE C2 and His6-RAGE VC1 deleted domains which have no ability to bind S100B peptides were applied. The proposed biosensor was quite sensitive, with a detection limit of 0.52 pM recorded in the buffer solution. The presence of diluted human plasma and 10 nM Aβ(1-40) have no influence on the biosensor performance.
在本研究中，我们报道了一种用于测定S100B蛋白的电化学生物传感器。用作分析活性分子的晚期糖基化终末产物（RAGE）受体的His标签化VC1结构域共价固定在沉积于金电极表面的与铜（II）形成的喷替酸（DPTA）复合物的硫醇衍生物单层上。RAGE VC1结构域与S100B蛋白之间的识别过程导致DPTA - Cu（II）中心的氧化还原活性发生变化，这通过奥斯特杨方波伏安法（OSWV）进行测量。为了验证观察到的分析信号是否源自His6 - RAGE VC1结构域与S100B蛋白之间的识别过程，使用了用无结合S100B肽能力的His6 - RAGE C2和His6 - RAGE VC1缺失结构域修饰的电极。所提出的生物传感器相当灵敏，在缓冲溶液中记录的检测限为0.52 pM。稀释的人血浆和10 nM Aβ(1 - 40)的存在对生物传感器性能没有影响。
