构建一种检测肺癌标志物神经元特异性烯醇化酶(neuron-specific enolase, NSE)的高性能电化学免疫传感器。

采用二维纳米材料氮化硼(boron nitride, BN)作为吸附基质沉积纳米金(aurum, Au)增加比表面积；二硫化钼负载铂钯(molybdenum disulfide@platinum-palladium, MoS₂@ PtPd)纳米模拟酶复合物用于电子转移催化，催化放大信号检测NSE；考察传感器构建关键因素，确定最优条件，进行传感器可重复性、稳定性和特异性试验及临床标本方法学验证。

BN直径200~500 nm，MoS₂直径500~800 nm，MoS₂@ PtPd复合物50~120 nm；电极表面结合BN电流下降40 μA，沉积金后电流增大15 μA，小牛血清白蛋白(bovine serum albumin, BSA)封闭电流减小9 μA，捕获NSE电流下降15μA；催化H₂O₂最佳总浓度为3μM，二抗与抗原结合最佳结合时间3 h；检测NSE标准曲线为I＝－5.13-0.40LogC_NSE，r＝0.92，最低检测限(low of detection, LOD)为0.86 pg/ml；传感器批间相对标准偏差(relative standard deviation, RSD)为2.91%，21 d后峰值电流保持其初始电流98.14%；1 ng/ml NSE与其他干扰蛋白对比，NSE(2.10±0.05)μA、甲胎蛋白(alpha-fetoprotein, AFP) (0.12±0.03)μA 、癌胚抗原(carcinoembryonic antigen, CEA) (0.15±0.02)μA、白蛋白(albumin, ALB) (0.21±0.02)μA、铁蛋白(ferritin, FER)(0.19±0.03)μA(P<0.05)；4例血清标本应用性相对偏差1.70%~4.11%，回收率93.09%~103.12%。

基于MoS₂@ PtPd催化联用BN/Au膜的新型免疫传感器用于NSE检测，构建简便、性能良好，可用于肺癌早期诊断，具有临床意义。

To develop a high-performance electrochemical immunosensor for detecting the lung cancer biomarker neuron specific enolase (NSE).

Two dimensional nano material boron nitride (BN) was used as the adsorption matrix to deposit nano gold (Au), effectively increasing the specific surface area; Using molybdenum disulfide supported platinum palladium (MoS₂@PtPd) nano mimetic enzyme complex for electron transfer catalysis and catalytic amplification signal detection of NSE; Determine the optimal conditions by examining the key factors involved in constructing sensors, and conduct repeatability, stability, and specificity tests on the sensors; And conduct methodological validation on clinical specimens.

It showes that the diameter range of BN was 200~500 nm, nm, the diameter range of MoS₂ was 500~800 nm, and the MoS₂ @ PtPd complex was about 50~120 nm. The current of BN binding on the electrode surface decreased by 40 μ A, and the current increased by 15 μ A after gold deposition. The blocking current of bovine serum albumin (BSA) decreased by 9 μ A, and the capture current of NSE decreased by 15 μ A; The optimal total concentration for catalyzing H₂O₂ is 3 μ M, and the optimal binding time for the secondary antibody to bind to the antigen is after 3 hours; When detecting NSE, the standard curve is: I=-5.13-0.40 LogC_NSE, r=0.92, and the low of detection (LOD) is 0.86 pg/ml; The relative standard deviation (R.S.D.) between sensor batches was 2.91%, and after 21 days, the peak current remained at 98.14% of its initial current; Compared with other interfering proteins at a concentration of 1 ng/ml, NSE was (2.10±0.05)μA, alpha fetoprotein (AFP) was (0.12±0.03)μA, carcinoembryonic antigen (CEA) was (0.15±0.02)μA, albumin (ALB) was (0.21±0.02)μA, and ferritin (FER) was (0.19±0.03)μA (P<0.05); The relative deviation of four serum samples with severe applicability ranges from 1.70% to 4.11%, and the recovery rate ranges from 93.09% to 103.12%.

The novel immunosensor based on MoS₂ @ PtPd catalytic combined with BN/Au membrane is simple to construct and has good performance for NSE detection. It provides an alternative method for early diagnosis of lung cancer, and has clinical significance.