Solanki Pratima R, Patel Manoj Kumar, Ali Md Azahar, Malhotra B D
Special Centre for Nanosciences, Jawaharlal Nehru University, New Delhi-110067, India.
J Mater Chem B. 2015 Aug 28;3(32):6698-6708. doi: 10.1039/c5tb00494b. Epub 2015 Jul 27.
We present a highly sensitive and selective electrochemical sandwich immunosensor (the analyte is "sandwiched" between two antibodies) based on chitosan (CH) modified nickel oxide (NiO) nanoparticles for the detection of Vibrio cholerae (Vc). The primary monoclonal antibodies specific to Vibrio cholerae (Ab-Vc) and bovine serum albumin (BSA) were co-immobilized on a CH-NiO surface deposited onto an indium tin oxide (ITO) coated glass electrode. The specific binding of Ab-Vc towards Vc was confirmed by interaction of secondary antibodies conjugated with protein [horse radish peroxidase (HRP)], with varying concentrations of hydrogen peroxide (HO), via electrochemical as well as optical techniques. The CH-NiO/ITO and Ab-Vc/CH-NiO/ITO electrodes have been characterized using X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy, high-resolution transmission electron microscopy and electrochemical techniques. This immunoelectrode (BSA/Ab-Vc/CH-NiO/ITO) exhibits a detection range of 20-700 ng mL with a sensitivity of 0.644 μA ng mL cm and a low detection range of 0.108 ng mL to Vc concentration. Besides this, the electrochemical response of the sandwich immunosensor (HRP-Ab-Vc/Vc/BSA/Ab-Vc/CH-NiO/ITO) towards HO concentration is found to be linear in the range of 10-50 mM with excellent sensitivity (2.95 mA mM cm).
我们展示了一种基于壳聚糖(CH)修饰的氧化镍(NiO)纳米颗粒的高灵敏度和选择性电化学夹心免疫传感器(分析物“夹”在两种抗体之间),用于检测霍乱弧菌(Vc)。将针对霍乱弧菌(Ab-Vc)和牛血清白蛋白(BSA)的一级单克隆抗体共同固定在沉积于氧化铟锡(ITO)涂层玻璃电极上的CH-NiO表面。通过与蛋白质[辣根过氧化物酶(HRP)]偶联的二级抗体与不同浓度的过氧化氢(HO)相互作用,利用电化学和光学技术证实了Ab-Vc与Vc的特异性结合。使用X射线衍射、傅里叶变换红外光谱、扫描电子显微镜、高分辨率透射电子显微镜和电化学技术对CH-NiO/ITO和Ab-Vc/CH-NiO/ITO电极进行了表征。这种免疫电极(BSA/Ab-Vc/CH-NiO/ITO)的检测范围为20 - 700 ng/mL,灵敏度为0.644 μA ng/mL cm,对Vc浓度的低检测限为0.108 ng/mL。除此之外,发现夹心免疫传感器(HRP-Ab-Vc/Vc/BSA/Ab-Vc/CH-NiO/ITO)对HO浓度的电化学响应在10 - 50 mM范围内呈线性,具有出色的灵敏度(2.95 mA mM/cm)。