Caro Nelson, Bruna Tamara, Guerreiro Antonio, Alvarez-Tejos Paola, Garretón Virginia, Piletsky Sergey, González-Casanova Jorge, Rojas-Gómez Diana, Ehrenfeld Nicole
Centro de Investigación AustralBiotech, Universidad Santo Tomas. Avenida Ejercito 146, Santiago 7591538, Chile.
Departament of Chemistry, University of Leincester, Leicester LE1 7RH, UK.
Nanomaterials (Basel). 2020 Feb 11;10(2):306. doi: 10.3390/nano10020306.
A simple and straightforward technique for coating microplate wells with molecularly imprinted polymer nanoparticles (nanoMIPs) to develop assays similar to the enzyme-linked immunosorbent (ELISA) assay to determine and quantify florfenicol (FF) in real food samples such as liquid milk and salmon muscle is presented here. The nanoMIPs were synthesized by a solid-phase approach with an immobilized FF (template) and characterized using dynamic light scattering, a SPR-2 biosensor system and transmission electron microscopy. Immobilization of nanoMIPs was conducted by preparing a homogenous solution of FF-nanoMIPs in water mixed with polyvinyl alcohol (PVA) 0.2% (w/v) in each well of a microplate. The detection of florfenicol was achieved in competitive binding experiments with a horseradish peroxidase-florfenicol (FF-HRP) conjugate. The assay made it possible to measure FF in buffer and in real samples (liquid milk and salmon muscle) within the range of 60-80 and 90-100 ng/mL, respectively. The immobilized nanoMIPs were stored for six weeks at room temperature and at 5 °C. The results indicate good signal recovery for all FF concentrations in spiked milk samples, without any detrimental effects to their binding properties. The high affinity of nanoMIPs and the lack of a requirement for cold chain logistics make them an attractive alternative to traditional antibodies used in ELISA.
本文介绍了一种简单直接的技术，用于在微孔板孔中包被分子印迹聚合物纳米颗粒（nanoMIPs），以开发类似于酶联免疫吸附测定（ELISA）的检测方法，用于测定和定量液态奶和三文鱼肌肉等实际食品样品中的氟苯尼考（FF）。通过固相方法以固定化的FF（模板）合成nanoMIPs，并使用动态光散射、SPR-2生物传感器系统和透射电子显微镜对其进行表征。通过在微孔板的每个孔中制备FF-nanoMIPs与0.2%（w/v）聚乙烯醇（PVA）的均匀水溶液来进行nanoMIPs的固定化。在与辣根过氧化物酶-氟苯尼考（FF-HRP）偶联物的竞争性结合实验中实现了氟苯尼考的检测。该检测方法能够分别在60 - 80 ng/mL和90 - 100 ng/mL的范围内测定缓冲液和实际样品（液态奶和三文鱼肌肉）中的FF。固定化的nanoMIPs在室温下和5℃下储存了六周。结果表明，加标牛奶样品中所有FF浓度的信号回收率良好，且对其结合特性没有任何不利影响。nanoMIPs的高亲和力以及对冷链物流的无需性使其成为ELISA中使用的传统抗体的有吸引力的替代品。
