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一种用于灵敏检测核基质蛋白22的高灵敏荧光免疫传感器,该蛋白作为膀胱癌早期诊断的生物标志物。

A highly sensitive fluorescent immunosensor for sensitive detection of nuclear matrix protein 22 as biomarker for early stage diagnosis of bladder cancer.

作者信息

Othman Hazha Omar, Salehnia Foad, Fakhri Neda, Hassan Rebwar, Hosseini Morteza, Faizullah Azad, Ganjali Mohammad Reza, Kazem Aghamir Seyed Mohammad

机构信息

Chemistry Department, College of Science, Salahaddin University-Erbil Iraq.

Center of Excellence in Electrochemistry, Faculty of Chemistry, University of Tehran Tehran 1417614418 Iran.

出版信息

RSC Adv. 2020 Aug 4;10(48):28865-28871. doi: 10.1039/d0ra06191c. eCollection 2020 Aug 3.

Abstract

A novel strategy is reported for highly sensitive, rapid, and selective detection of nuclear matrix protein NMP22 using two-color quantum dots based on fluorescence resonance energy transfer (FRET). Quantum dots (QDs) are highly advantageous for biological imaging and analysis, particularly when combined with (FRET) properties of semiconductor quantum dot (QDs) are ideal for biological analysis to improve sensitivity and accuracy. In this FRET system narrowly dispersed green emitting quantum dot CdTe core is used as a donor and labelled by monoclonal (mAb) antibody, while orange emitting quantum dot CdTe/CdS core shell is used as an accepter and labelled by polyclonal (pAb) antibody. The quantum dots are labelled by antibodies using EDC/NHS as crosslinking agent. Bovine serum albumin (BSA) solution was added to block nonspecific binding sites. The fluorescence intensity of QDs accepter decreased linearly with the increasing concentrations of NMP22 from 2-22 pg mL due to FRET system and fluoroimmunoassay reaction. This method has good regression coefficient ( = 0.998) and detection limit was 0.05 pg mL. The proposed FRET-based immunosensor provides a quick, simple and sensitive immunoassay tool for protein detection, and can be considered as a promising approach for clinical applications. The proposed FRET-based immunosensor provides a quick, simple and sensitive immunoassay tool for protein detection, and can be considered as a promising approach for clinical applications.

摘要

报道了一种基于荧光共振能量转移(FRET)的双色量子点高灵敏度、快速且选择性检测核基质蛋白NMP22的新策略。量子点(QDs)在生物成像和分析方面具有高度优势,特别是当与半导体量子点的(FRET)特性相结合时,非常适合用于生物分析以提高灵敏度和准确性。在这个FRET系统中,窄分散的绿色发射量子点CdTe核用作供体并由单克隆(mAb)抗体标记,而橙色发射量子点CdTe/CdS核壳用作受体并由多克隆(pAb)抗体标记。使用EDC/NHS作为交联剂通过抗体对量子点进行标记。加入牛血清白蛋白(BSA)溶液以封闭非特异性结合位点。由于FRET系统和荧光免疫分析反应,量子点受体的荧光强度随着NMP22浓度从2 - 22 pg/mL增加而呈线性下降。该方法具有良好的回归系数( = 0.998),检测限为0.05 pg/mL。所提出的基于FRET的免疫传感器为蛋白质检测提供了一种快速、简单且灵敏的免疫分析工具,可被视为临床应用的一种有前景的方法。所提出的基于FRET的免疫传感器为蛋白质检测提供了一种快速、简单且灵敏的免疫分析工具,可被视为临床应用的一种有前景的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d4d6/9055858/ab31e7008df5/d0ra06191c-f1.jpg

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