Key Laboratory of Analysis and Detection for Food Safety (Ministry of Education of China and Fujian Province), Department of Chemistry, Fuzhou University, Fuzhou 350108, PR China.
Biosens Bioelectron. 2011 Jan 15;26(5):2090-6. doi: 10.1016/j.bios.2010.09.012. Epub 2010 Sep 16.
A sensitive electrochemical immunosensor for the fast screening of brevetoxin B (BTX-2) in food samples was developed by means of immobilizing BTX-2-bovine serum albumin conjugate (BTX-2-BSA) on the gold nanoparticles-decorated amine-terminated poly(amidoamine) dendrimers (AuNP-PAADs). The presence of gold nanoparticles greatly improved the conductivity of the PAADs, and three-dimensional PAADs increased the surface coverage of the biomolecules on the electrode. Under optimal conditions, three types of immunosensor, i.e. with AuNPs, PAADs, or AuNP-PAADs, were used for the determination of BTX-2 in a competitive-type immunoassay format using horseradish peroxidase-labeled anti-BTX antibodies (HRP-anti-BTX-2) as trace in the H(2)O(2)-o-phenylenediamine (o-PD) system. A low detection limit (LOD) of 0.01 ng/mL and a wide dynamic working linear range of 0.03-8 ng/mL BTX-2 using AuNP-PAADs as matrices were obtained in comparison with those of only using AuNP or PAADs. Intra-batch assay precision was substantially improved by resorting to the AuNP-PAADs manifold. The proposed method features unbiased identification of negative (blank) and positive samples. No significant differences were encountered in the analysis of the spiking real samples between the electrochemical immunosensor and liquid chromatography for the determination of BTX-2. Importantly, this method provided a biocompatible immobilization and a promising immunosensing platform for analytes with small molecules in the analysis and detection of food safety.
一种用于快速筛选食品样品中布雷毒素 B(BTX-2)的灵敏电化学免疫传感器,是通过将 BTX-2-牛血清白蛋白结合物(BTX-2-BSA)固定在金纳米粒子修饰的胺端基聚(酰胺-胺)树枝状聚合物(AuNP-PAADs)上来制备的。金纳米粒子的存在极大地提高了 PAADs 的导电性,而三维 PAADs 增加了电极上生物分子的表面覆盖率。在最佳条件下,使用三种类型的免疫传感器,即带有 AuNPs、PAADs 或 AuNP-PAADs,在使用辣根过氧化物酶标记的抗-BTX 抗体(HRP-anti-BTX-2)作为示踪物的竞争型免疫分析格式中,用于测定 H 2 O 2 -邻苯二胺(o-PD)系统中的 BTX-2。与仅使用 AuNP 或 PAADs 相比,使用 AuNP-PAADs 作为基质,BTX-2 的检测下限(LOD)为 0.01 ng/mL,动态工作线性范围较宽,为 0.03-8 ng/mL BTX-2。通过使用 AuNP-PAADs 作为多重分析,大大提高了批内测定的精密度。该方法具有对阴性(空白)和阳性样品进行无偏识别的特点。在使用电化学免疫传感器和液相色谱法测定 BTX-2 时,对实际加标样品的分析没有发现明显差异。重要的是,该方法为小分子分析物的分析和检测提供了一种具有生物相容性的固定化和有前途的免疫传感平台。