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基于鸟嘌呤功能化石墨烯纳米带的磁控电化学免疫分析在贝类中检测膝沟藻毒素 B。

Magneto-controlled electrochemical immunoassay of brevetoxin B in seafood based on guanine-functionalized graphene nanoribbons.

机构信息

Ministry of Education & Fujian Provincial Key Laboratory of Analysis and Detection for Food Safety, Department of Chemistry, Fuzhou University, Fuzhou 350108, PR China.

出版信息

Biosens Bioelectron. 2012 Oct-Dec;38(1):86-93. doi: 10.1016/j.bios.2012.05.006. Epub 2012 May 24.

Abstract

A facile and feasible magneto-controlled immunosensing platform was designed for sensitive electrochemical immunoassay of brevetoxin B (BTX-2) in seafood by using guanine-assembled graphene nanoribbons (GGNRs) as molecular tags on a home-made magnetic carbon paste electrode. Initially, monoclonal mouse anti-BTX-2 antibodies were covalently immobilized on the surface of magnetic beads, which were used as the immunosensing probes for the capture of BTX-2. The recognition elements were prepared by chemical modification of bovine serum albumin-BTX-2 conjugates (BTX-2-BSA) with the GGNRs. Based on a competitive-type immunoassay format, the formed magnetic immunocomplex was integrated on the electrode with an external magnet, followed by determination in pH 6.5 phosphate-buffered solution containing 2 μM Ru(bpy)(3)Cl(2). Under optimal conditions, the electrochemical signals decreased with the increasing BTX-2 concentrations in the sample. The dynamic concentration range spanned from 1.0 pg mL(-1) to 10 ng mL(-1) with a detection limit of 1.0 pg mL(-1) BTX-2. Inter- and intra-batch assay precisions were substantially improved by resorting to the GGNR manifold. The method featured unbiased identification of negative (blank) and positive samples. No significant differences at the 95% confidence level were encountered in the analysis of 12 spiked samples including S. constricta, M. senhousia and T. granosa between the electrochemical immunoassay and commercially available enzyme-linked immunosorbent assay (ELISA) for determination of BTX-2.

摘要

设计了一种简便可行的磁控免疫传感平台,用于通过使用鸟嘌呤组装的石墨烯纳米带(GGNRs)作为分子标记物,在自制的磁性碳糊电极上对海鲜中的布雷毒素 B(BTX-2)进行灵敏的电化学免疫分析。最初,单克隆抗 BTX-2 抗体被共价固定在磁性珠的表面上,用作捕获 BTX-2 的免疫传感探针。识别元件是通过牛血清白蛋白-BTX-2 缀合物(BTX-2-BSA)与 GGNRs 的化学修饰制备的。基于竞争型免疫分析格式,形成的磁性免疫复合物在外加磁场的作用下被集成在电极上,随后在含有 2 μM Ru(bpy)(3)Cl(2)的 pH 6.5 磷酸盐缓冲溶液中进行测定。在最佳条件下,电化学信号随样品中 BTX-2 浓度的增加而降低。动态浓度范围从 1.0 pg mL(-1)到 10 ng mL(-1),检测限为 1.0 pg mL(-1)BTX-2。通过采用 GGNR 歧管,可以大大提高批间和批内测定的精密度。该方法具有对阴性(空白)和阳性样品进行无偏识别的特点。在分析 12 个包括 S. constricta、M. senhousia 和 T. granosa 在内的加标样品时,电化学免疫分析与商业酶联免疫吸附测定(ELISA)之间在 BTX-2 的测定中未发现 95%置信水平的显著差异。

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