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通过银离子交换反应在CdTe量子点浸渍纸上进行的距离依赖性视觉荧光免疫分析。

Distance-dependent visual fluorescence immunoassay on CdTe quantum dot-impregnated paper through silver ion-exchange reaction.

作者信息

Huang Lingting, Wang Jun, Wang Qingshui, Tang Dianping, Lin Yao

机构信息

Central Laboratory at The Second Affiliated Hospital of Fujian Traditional Chinese Medical University, Collaborative Innovation Center for Rehabilitation Technology, Fujian University of Traditional Chinese Medicine, Fuzhou, 350122, Fujian, People's Republic of China.

Key Laboratory for Analytical Science of Food Safety and Biology (MOE & Fujian Province), Department of Chemistry, Fuzhou University, Fuzhou, 350108, People's Republic of China.

出版信息

Mikrochim Acta. 2020 Sep 12;187(10):563. doi: 10.1007/s00604-020-04546-7.

Abstract

A paper-based visual fluorescence immunoassay is presented for the detection of matrix metalloproteinase-7 (MMP7) that is related to renal cancer. The method is based on the distance-dependent fluorescence quenching of CdTe quantum dots (QDs) on a nitrocellulose membrane by Ag following a sandwich-type immunoreaction on microtiter wells using silver nanoparticle (AgNP)-labeled secondary antibody- and primary antibody-coated microtiter wells. The silver nanoparticles captured in the well are dissolved with HNO, while the quenching effect of QDs is based on silver ion-exchange reaction under 365-nm excitation light irradiation. Increasing concentration of released Ag, thus higher concentration of the protein, leads to an increased distance of quenching on the nitrocellulose membrane. The paper-based immunoassay by combination of AgNP-assisted ion-exchange reaction with QD gives good distance-dependent responses and allows the detection of MMP7 at a concentration as low as 7.3 pg mL. The coefficients of variation are less than 6.9% and 12.4% for intra-assay and inter-assay, respectively. High specificity and long-term stability are achieved during the assay. Importantly, the testing of human serum samples using our strategy shows well-matched results with commercial human MMP7 ELISA kits. Graphical abstract A distance-dependent visual immunoassay is developed for the determination of serum matrix metalloproteinase-7 on CdTe quantum dot-impregnated paper with silver ion-exchange reaction.

摘要

本文提出了一种基于纸质的视觉荧光免疫分析法,用于检测与肾癌相关的基质金属蛋白酶-7(MMP7)。该方法基于在微孔板上进行夹心型免疫反应后,硝酸纤维素膜上的碲化镉量子点(QDs)因银而产生的距离依赖性荧光猝灭,其中微孔板用银纳米颗粒(AgNP)标记的二抗和一抗包被。微孔中捕获的银纳米颗粒用硝酸溶解,而量子点的猝灭效应基于365 nm激发光照射下的银离子交换反应。释放的银浓度增加,即蛋白质浓度升高,会导致硝酸纤维素膜上猝灭距离增加。通过将AgNP辅助离子交换反应与量子点相结合的基于纸质的免疫分析法具有良好的距离依赖性响应,能够检测低至7.3 pg/mL浓度的MMP7。批内和批间变异系数分别小于6.9%和12.4%。该分析过程具有高特异性和长期稳定性。重要的是,使用我们的策略对人血清样本进行检测,结果与商用人类MMP7 ELISA试剂盒的结果高度匹配。图形摘要 开发了一种基于距离依赖性的视觉免疫分析法,用于通过银离子交换反应在浸渍碲化镉量子点的纸上测定血清基质金属蛋白酶-7。

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