Facile engineered polymeric microdevice via co-coupling of phenylboronic acid and Protein A for oriented antibody immobilization enables substantial signal enhancement for an enhanced fluorescence immunoassay
Facile engineered polymeric microdevice via co-coupling of phenylboronic acid and Protein A for oriented antibody immobilization enables substantial signal enhancement for an enhanced fluorescence immunoassay
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DOI：
10.1016/j.snb.2021.130444
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发表时间：
2021-07-23
影响因子：
8.4
通讯作者：
Li, Lu
中科院分区：
文献类型：
--
作者：
Chang, Jianguo;Gao, Nailong;Li, Lu
强大的免疫分析法在检测微量物质方面越来越受到重视。本研究基于苯硼酸与蛋白A共偶联的定向抗体固定化方法，提出了一种基于聚合装置的荧光免疫分析方法，提高了甲胎蛋白（AFP）分析的检测性能。与传统的物理吸收或化学共价偶联固定抗体的方法不同，在本免疫测定中，定向固定方式使AFP抗体获得了最大的生物反应性。在聚甲基丙烯酸甲酯（PMMA）表面，由于共偶格相关距离效应，AFP抗体的包封密度显著提高。两性离子2-甲基丙烯酰氧乙基磷胆碱（MPC）与牛血清白蛋白（BSA）联合使用可显著抑制其非特异性吸收。这些综合特征有助于在AFP存在时输出强荧光，而在AFP不存在时发出非常弱的背景。经过典型的夹心免疫反应后，免疫传感器记录的荧光强度与甲胎蛋白浓度成正比，对甲胎蛋白检测具有良好的灵敏度、特异性和可靠性。对比研究表明，共耦合优于单耦合。我们设想这种独特的免疫测定方法可以为在疾病诊断、食品安全检测和环境监测领域建立免疫传感器提供新的途径。
The powerful immunoassay has attracted increasing attention for detecting trace substances. Herein, based on the co-coupling of phenylboronic acid and Protein A for oriented antibodies immobilization, we proposed a polymeric device-based fluorescence immunoassay with enhanced assay performance for Alpha-fetoprotein (AFP) analysis. Unlike traditional physical absorption or chemical covalent conjugation to immobilize antibodies, in this immunoassay, the bioreactivity of AFP antibody was maximumly obtained owing to the oriented immobilization manner. The package density of AFP antibody on the polymethylmethacrylate (PMMA) surface is significantly improved because of the co-coupling case associated distance effect. In addition, the non-specific absorption is greatly inhibited with the combined using of zwitterionic 2-methacryloyloxyethyl phosphorylcholine (MPC) and bovine serum albumin (BSA). These integrated features facilitate the output of strong fluorescence in the presence of AFP but emit very weak background in the absence of AFP. After a typical sandwich immunoreaction, the recorded fluorescence intensity of the immunosensor is proportional to the AFP concentration, showing excellent sensitivity, specificity, and reliability for AFP determination. Comparative studies show the co-coupling is superior than its single coupling counterparts. We envision this unique immunoassay can provide a new avenue for building immunosensors in the fields of disease diagnosis, food safety detection, and environmental monitoring.