中文摘要：

本研究在玻碳电极（GCE）表面电沉积金纳米粒子（Au NPs）,通过化学吸附将微囊藻毒素-（亮氨酸-精氨酸）（MC-LR）的单克隆抗体（anti-MC-LR）固定在电沉积了Au NPs的玻碳电极表面,以牛血清白蛋白（BSA）封闭非特异性吸附位点,制得免疫电极anti-MC-LR/Au NPs/GCE。采用微乳化法制备了掺杂三（2,2＇-联二吡啶）钌（Ⅱ）配合物离子（Ru（bpy）2＋3）的二氧化硅纳米粒子（Ru@SiO2）,利用透射电镜和扫描电镜对所制备的纳米粒子进行表征。3-氨基丙基三乙氧基硅烷（APTS）进一步与Ru@SiO2反应,制得氨基功能化的Ru@SiO2,通过1-（3-二甲氨基丙基）-3-乙基碳二亚胺（EDC）和N-羟基琥珀酰亚胺（NHS）活化辣根过氧化物酶标记的MC-LR（HRP-MC-LR）,并使其与氨基功能化的Ru@SiO2偶联,制得MC-LR-Ru@SiO2。采用直接竞争模式,在标记物MC-LR-Ru@SiO2存在下,以三丙胺作为共反应物,利用电化学发光法（ECL）测定溶液中的微囊藻毒素,免疫反应完成后,电化学发光强度（I）随着MC-LR浓度的增大而减小,且在0.100~100μg/L范围内,电化学发光强度差值（ΔI）与游离的MC-LR浓度的对数呈良好线性关系,检出限为0.007μg/L。对实际水样进行了加标回收实验,回收率为95.5%~105%。

英文摘要：

Gold nanoparticles（ Au NPs） were electrodeposited onto the surface of a glassy carbon electrode（ GCE）. The antibody of microcystin-（ leucine-arginine）（ anti-MC-LR） was immobilized on the film surface of Au NPs by the adsorption between Au nanoparticles and antibody. Non-specific adsorption sites on the Au NPs were blocked with bovine serum albumin（ BSA） to prepare an immunoelectrode anti-MC-LR / Au NPs / GCE. Microemulsion polymerization method was adopted to synthesize silica nanoparticles doped with tris（ 2,2＇-bipyridyl） ruthenium（ Ⅱ） complex ions（ e. g. Ru@SiO2）. Ru@ SiO2 nanoparticles and Au NPs on the surface of a GCE were characterized by transmission electron microscopy image（ TEM） and scanning electron microscopy（ SEM）,respectively. The Ru@ SiO2 nanoparticles were further modified with amino groups by reacting with 3-aminopropyltriethoxysilane. The amino group modified Ru@ SiO2 was then crosslinked with the carboxylic groups of microcystin-（ leucine-arginine）（ MC-LR） by ethyl-3-（ dimethyl aminopropyl） carbodiimide（ EDC）and N-hydroxysuccinimide（ NHS） to prepare Ru@SiO2-conjugated MC-LR（ MC-LR-Ru @SiO2）. The immunosensor was based on the direct competitive immunoassay format between the labeled agent of MC-LR-Ru @ SiO2 and the analyte. In the presence of MC-LR-Ru@SiO2,tripropylamine（ TPA） was used as coreactant. MC-LR in solution was determined by electrochemiluminescence（ ECL） method. The ECL intensity（ I） decreases with the increase of MC-LR concen-tration after antigen-antibody reaction. A good linear relationship between the change of ECL intensity（ ΔI） and the logarithm of MC-LR concentration was obtained in the range of 0. 100-100 μg / L with a detection limit of 0. 007 μg / L. The developed ECL immunosensor was used to determine MC-LR in real water samples with recoveries of 95. 5%-105%.