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采用分子印迹光子聚合物涂层膜和光纤光谱仪对 S 层蛋白进行选择性、灵敏性和快速测定。

Selective, sensitive, and fast determination of S-layer proteins by a molecularly imprinted photonic polymer coated film and a fiber-optic spectrometer.

机构信息

State Key Laboratory for Quality and Safety of Agro-products, Ningbo University, Ningbo, Zhejiang, 315211, China.

School of Food and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210023, Jiangsu, China.

出版信息

Anal Bioanal Chem. 2019 Nov;411(29):7737-7745. doi: 10.1007/s00216-019-02109-1. Epub 2019 Nov 11.

Abstract

A newly developed molecularly imprinted photonic polymer (MIPP) film, which was prepared by colloidal crystal templating and surface molecular imprinting, was used for selective capture of S-layer protein (SLP) from a complex Lactobacillus acidophilus sample. The colloidal crystal templates were formed by a dipping process followed by chemical binding of the imprinted template SLP molecules. A sandwich structure consisting of two glass slides was formed after the SLP-silica layer had been covered with a poly(methyl methacrylate) glass slide. After polymerization of the SLP-silica layer with the preprepared polymerization solution, hydrofluoric acid and acetic phosphate buffer solutions removed the silica particles and SLP molecules, respectively. The MIPP film obtained exhibited a three-dimensional, highly ordered and interconnected macroporous structure (pore size greater than 200 nm), which is specifically accessible to SLP molecules. The adsorbed SLP molecules were simply and straightforwardly detected by a fiber-optic spectrometer. The redshift of the Bragg diffraction peak of the MIPP film was linearly related to the number of SLP molecules that had been harvested in the film. The detection limit of the SLP-MMIP-fiber-optic spectrometer method for SLP was 1 ng mL. The MIPP sensor was successfully applied to detect SLP molecules in a crudely extracted Lactobacillus acidophilus sample. Our results prove the applicability of the SLP-MIPP film for fast and real-time measurement of SLP. Graphical abstract.

摘要

一种新开发的分子印迹光子聚合物(MIPP)薄膜,通过胶体晶体模板和表面分子印迹制备,用于从复杂的嗜酸乳杆菌样品中选择性捕获 S-层蛋白(SLP)。胶体晶体模板通过浸渍工艺形成,随后通过化学结合印迹模板 SLP 分子。在 SLP-二氧化硅层被聚甲基丙烯酸甲酯玻璃片覆盖后,形成了由两个玻璃片组成的夹层结构。在用预制备的聚合溶液聚合 SLP-二氧化硅层后,氢氟酸和乙酸磷酸盐缓冲溶液分别去除了二氧化硅颗粒和 SLP 分子。所得 MIPP 薄膜呈现出三维、高度有序且相互连接的大孔结构(孔径大于 200nm),专门可容纳 SLP 分子。吸附的 SLP 分子可以通过光纤光谱仪简单直接地检测到。MIPP 薄膜的布拉格衍射峰的红移与已在薄膜中收获的 SLP 分子数量呈线性关系。用于 SLP 的 SLP-MMIP-光纤光谱仪方法的检测限为 1ng mL。MIPP 传感器成功应用于检测粗提嗜酸乳杆菌样品中的 SLP 分子。我们的结果证明了 SLP-MIPP 薄膜适用于快速实时测量 SLP。

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