Florfenicol Binding to Molecularly Imprinted Polymer Nanoparticles in Model and Real Samples
Florfenicol Binding to Molecularly Imprinted Polymer Nanoparticles in Model and Real Samples
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DOI：
10.3390/nano10020306
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发表时间：
2020-02-01
期刊：
影响因子：
5.3
通讯作者：
Ehrenfeld, Nicole
中科院分区：
文献类型：
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作者：
Caro, Nelson;Bruna, Tamara;Ehrenfeld, Nicole
介绍了一种用分子印迹聚合物纳米颗粒(NanMIP)包覆微孔板的简单、直接的技术，以建立类似于酶联免疫吸附(ELISA)分析的分析方法，以检测和定量真实食品样品中的氟苯尼考(FF)，如液态牛奶和鲑鱼肌肉。采用固相法制备了纳米分子印迹聚合物，并用动态光散射、SPR-2生物传感器系统和透射电子显微镜对其进行了表征。纳米分子印迹聚合物的固定化是通过在微孔板的每孔中制备FF-纳米分子印迹聚合物掺入0.2%(w/v)聚乙烯醇(PVA)的水中的均一溶液来实现的。通过与辣根过氧化物酶氟苯尼考(FF-HRP)的竞争结合实验，实现了对氟苯尼考的检测。该方法可以测定缓冲液和实际样品(液态奶和鲑鱼肌肉)中的FF，范围分别为60~80和90~100 ng/m L。固定化的纳米MIP在室温和5℃下保存6周。结果表明，对于添加牛奶样品中的所有FF浓度，信号恢复良好，对其结合性能没有任何不利影响。纳米MIP的高亲和力和不需要冷链物流的特点使其成为一种有吸引力的替代传统抗体用于ELISA的方法。
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 degrees 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.