Departamento de Bioquímica, Universidade Federal de Pernambuco-UFPE, 50670-901 Recife, PE, Brazil.
J Colloid Interface Sci. 2011 Oct 1;362(1):194-201. doi: 10.1016/j.jcis.2011.06.042. Epub 2011 Jun 24.
We report the development of a new selective and specific electrochemical biosensor for bacterial lipolysaccharide (LPS). An electrode interface was constructed using a l-cysteine-gold nanoparticle (AuNpCys) composite to be immobilized by electrostatic interaction in the network of a poly(vinyl chloride-vinyl acetate maleic acid) (PVM) layer on a gold bare electrode. The impedimetric biosensor is fabricated by self-assembled CramoLL lectin on the PVM-AuNpCys-modified gold electrode through electrostatic interaction. CramoLL is used as the recognition interface. AFM images showed that LPS was specifically recognized on the PVM-AuNpCys-CramoLL system surface. The measurements of cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) showed that the electrochemical response of a redox probe system (K(4)Fe(CN)(6)/K(3)Fe(CN)(6)) were blocked, due to the procedures of modified electrode with PVM-AuNpCys-CramoLL. In the majority of the experiments the lectin retained its activity as observed through its interaction with LPS from Escherichia coli, Serratia marcescens, Salmonella enterica and Klebsiella pneumoniae. The results are expressed in terms of the charge transfer resistance and current peak anodic using the EIS and CV techniques for the development of a biosensor for contamination by endotoxins. A new type of sensor for selective discrimination of LPS types with a high sensitivity has been obtained.
我们报告了一种新的选择性和特异性电化学生物传感器的开发,用于细菌脂多糖(LPS)。使用巯基-L-半胱氨酸-金纳米粒子(AuNpCys)复合材料通过静电相互作用在聚(氯乙烯-醋酸乙烯酯-马来酸)(PVM)层的网络中构建电极界面,该层固定在金裸电极上。通过静电相互作用,将自组装的 CramoLL 凝集素固定在 PVM-AuNpCys 修饰的金电极上,制备了阻抗生物传感器。CramoLL 用作识别界面。原子力显微镜图像表明,LPS 特异性地识别在 PVM-AuNpCys-CramoLL 系统表面。循环伏安法(CV)和电化学阻抗谱(EIS)的测量表明,由于用 PVM-AuNpCys-CramoLL 修饰电极的过程,氧化还原探针系统(K(4)[Fe(CN)(6)](4-)/ K(3)[Fe(CN)(6)](3-))的电化学响应被阻断。在大多数实验中,凝集素保留了与大肠杆菌、粘质沙雷氏菌、肠炎沙门氏菌和肺炎克雷伯菌的 LPS 相互作用的活性。结果以使用 EIS 和 CV 技术开发用于内毒素污染的生物传感器的电荷转移电阻和阳极电流峰的形式表示。已经获得了一种用于 LPS 类型的高灵敏度选择性区分的新型传感器。