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基于 Au 星@BSA-鲁米诺纳米复合材料的夹心型 ECL 免疫传感器用于测定人绒毛膜促性腺激素。

Sandwich-format ECL immunosensor based on Au star@BSA-Luminol nanocomposites for determination of human chorionic gonadotropin.

机构信息

Institute of Nano Biomedicine and Engineering, Shanghai Engineering Research Center for Intelligent Instrument for Diagnosis and Therapy, Department of Instrument Science & Engineering, School of Electronic Information and Electrical Engineering, Shanghai Jiao Tong University, 800 Dongchuan RD, Shanghai 200240, PR China.

The Education Ministry Key Laboratory of Resource Chemistry, Shanghai Key Laboratory of Rare Earth Functional Materials and Shanghai Municipal Education Committee Key Laboratory of Molecular Imaging Probes and Sensors, Department of Chemistry, Shanghai Normal University, Shanghai 200234, PR China.

出版信息

Biosens Bioelectron. 2018 Mar 15;101:219-226. doi: 10.1016/j.bios.2017.10.040. Epub 2017 Oct 18.

Abstract

A sandwich-configuration electrochemiluminescence (ECL) immunosensor based on Au star@BSA-Luminol nanocomposites for ultrasensitive determination of human chorionic gonadotropin (HCG) has been developed. In this work, nanostructured Polyaniline hydrogels (Pani) decorated with Pt nanoparticles (Pani/Pt) were utilized to construct the base of this immunosensor, greatly increasing the amount of loaded capture antibodies (Ab) via linkage reagent glutaraldehyde (GA). The used conducting Pani/Pt nanocomposites possessed the unique features, such as large surface area, high electron transfer speed and favorable electrocatalytic activities of hydrogen peroxide, which offered a prominent platform for this sandwich-sensor and acted as efficient ECL signal amplifier also. Furthermore, we employed horseradish peroxidase (HRP) to block the nonspecific binding sites instead of commonly used bovine serum albumin (BSA), which further amplified the signal of luminol in the present of hydrogen peroxide (HO). In addition, Au star@BSA nanocomposites with excellent water solubility, low-toxicity and great biocompatibility were prepared and used to immobilize HCG detection antibodies (Ab) and luminescent material luminol. Then, the above-synthetized Luminol-Au star@BSA-Ab complex was attached to the modified sensor by sandwiched immunoreactions. Under the optimized conditions, the proposed immunosensor exhibited a sensitive detection of HCG in a wide linear range from 0.001 to 500mIUmL with a detection limit of 0.0003mIU/mL (S/N = 3). All the results indicated that such a sandwiched HCG immunosensor exhibited favorable ECL analytical performance. This developed method may be potentially used to recognize other clinical protein and display a novel ideal to construct an immunosensor.

摘要

基于金纳米星@BSA-鲁米诺纳米复合材料的三明治结构电化学发光(ECL)免疫传感器已被开发出来,用于超灵敏测定人绒毛膜促性腺激素(HCG)。在这项工作中,用纳米结构聚苯胺水凝胶(Pani)修饰的铂纳米粒子(Pani/Pt)构建了这个免疫传感器的基底,通过连接试剂戊二醛(GA)大大增加了负载的捕获抗体(Ab)的量。所使用的导电 Pani/Pt 纳米复合材料具有独特的特性,如大的表面积、高电子转移速度和对过氧化氢的有利电催化活性,这为三明治传感器提供了一个突出的平台,同时也作为有效的 ECL 信号放大器。此外,我们采用辣根过氧化物酶(HRP)代替常用的牛血清白蛋白(BSA)来封闭非特异性结合位点,这进一步放大了在过氧化氢(HO)存在下鲁米诺的信号。此外,制备了具有优异水溶性、低毒性和良好生物相容性的 Au 纳米星@BSA 纳米复合材料,并用于固定 HCG 检测抗体(Ab)和发光材料鲁米诺。然后,通过夹心免疫反应将上述合成的鲁米诺-Au 纳米星@BSA-Ab 复合物固定在修饰后的传感器上。在优化条件下,该免疫传感器对 HCG 的检测表现出宽线性范围(0.001-500mIUmL),检测限为 0.0003mIU/mL(S/N = 3)。所有结果表明,这种夹心 HCG 免疫传感器表现出良好的 ECL 分析性能。这种开发的方法可能有潜力用于识别其他临床蛋白质,并为构建免疫传感器提供一种新的理想方法。

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