Magnetite-Quantum Dot Immunoarray for Plasmon-Coupled-Fluorescence Imaging of Blood Insulin and Glycated Hemoglobin.
Magnetite-Quantum Dot Immunoarray for Plasmon-Coupled-Fluorescence Imaging of Blood Insulin and Glycated Hemoglobin.
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DOI：
10.1021/acssensors.7b00124
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发表时间：
2017-07-28
期刊：
影响因子：
8.9
通讯作者：
Krishnan S
中科院分区：
文献类型：
--
作者：
Singh V;Nerimetla R;Yang M;Krishnan S
新的微阵列芯片策略具有灵敏性和选择性，可以直接测量血液样本中低水平的重要生物标志物，通过及时诊断异常状况，对于改善人类健康具有重要意义。在此，我们设计了一种抗体-适体免疫阵列芯片，以证明使用表面等离子体微阵列同时测量与糖尿病和糖尿病前期疾病相关的血液胰岛素和糖化血红蛋白（HbA1c）水平，并通过荧光成像进行验证。为了在同一样品上完成表面等离子体和荧光成像，我们用量子点装饰磁铁矿纳米粒子，用于共价固定适体，以便随后从稀释 20 倍的全血样品中捕获和分离胰岛素和 HbA1c 标记物特异性的适体。这种新的免疫阵列平台实现了直接的临床相关分析以及两种标记物的荧光成像。胰岛素的检测限为 4 pM，HbA1c 的检测限为 1%。使用凝血酶和血小板衍生生长因子进行的串扰检查证实，设计的免疫阵列对胰岛素和 HbA1c 具有高度选择性。表面等离子体动力学分析提供了胰岛素和 HbA1c 与其表面固定化单克隆抗体结合的表观结合常数，分别为 0.24 (±0.08) nM 和 37 (±3) μM。因此，在设计的免疫阵列芯片中，独特地获得了具有结合动力学的超低水平血液生物标志物水平的定量成像。总之，本报告证明了所开发的磁铁矿-量子点-生物共轭策略对于具有分子结合相互作用特征的全血生物标志物的临床诊断具有重要意义。
New microarray chip strategies that are sensitive and selective and that can measure low levels of important biomarkers directly in a blood sample are significant for improving human health by allowing timely diagnosis of an abnormal condition. Herein, we designed an antibody–aptamer immunoarray chip to demonstrate simultaneous measurement of blood insulin and glycated hemoglobin (HbA1c) levels relevant to diabetic and prediabetic disorders using a surface plasmon microarray with validation by fluorescence imaging. To accomplish both surface plasmon and fluorescence imaging on the same sample, we decorated magnetite nanoparticles with quantum dots for covalent immobilization of aptamers for subsequent capture and isolation of the aptamers specific for insulin and HbA1c markers from 20-times diluted whole blood samples. Direct clinically relevant analysis, along with fluorescent imaging of the two markers, was achieved by this new immunoarray platform. The limit of detection was 4 pM for insulin and 1% for HbA1c. Examination of cross-talk using thrombin and platelet-derived growth factor confirmed that the designed immunoarray was highly selective for insulin and HbA1c. Surface plasmon kinetic analysis provided apparent binding constants of 0.24 (±0.08) nM and 37 (±3) μM, respectively, for the binding of insulin and HbA1c onto their surface immobilized monoclonal antibodies. Thus, quantitative imaging of ultralow levels of blood biomarker levels with binding kinetics is uniquely obtained in the designed immunoarray chip. In conclusion, this report demonstrates considerable significance of the developed magnetite-quantum dot-bioconjugate strategy for clinical diagnostics of whole blood biomarkers with characterization of molecular binding interactions.
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影响因子：
9.8
作者：
GOETZ, FC;FRENCH, LR;CLEMENTS, JP
通讯作者：
CLEMENTS, JP
影响因子：
16.2
作者：
American Diabetes Association
通讯作者：
American Diabetes Association
影响因子：
10.2
作者：
Ahmed S;Dong J;Yui M;Kato T;Lee J;Park EY
通讯作者：
Park EY
影响因子：
16.6
作者：
Krishnan, Sadagopan;Mani, Vigneshwaran;Wasalathanthri, Dhanuka;Kumar, Challa V.;Rusling, James F.
通讯作者：
Rusling, James F.
影响因子：
4.3
作者：
Boonyasit, Yuwadee;Heiskanen, Arto;Laiwattanapaisal, Wanida
通讯作者：
Laiwattanapaisal, Wanida