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首页> 外文期刊>Applied Surface Science >Thin solid europium(III) dye layers as donors in time-resolved fluorescence resonance energy transfer assays
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Thin solid europium(III) dye layers as donors in time-resolved fluorescence resonance energy transfer assays

机译：薄固体solid（III）染料层作为时间分辨荧光共振能量转移测定中的供体

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摘要

Lanthanide chelates and lanthanide nanoparticle labels are attractive donors for separation-free time-resolved fluorescence resonance energy transfer (TR-FRET) assays. In fully dyed nanoparticles, the inner volume of nanoparticle labels in TR-FRET assays are incapable of participating to energy transfer due to large distances to acceptors on the surface. Our interest was to study surface-based TR-FRET and, therefore, various europium(III) (Eu) chelate layers were investigated for TR-FRET efficiency. Eu(III) chelates incorporated in a siloxane layer, Eu(III) chelate covalently coupled on silanized surface and Eu(III) labeled protein surface were prepared and compared to nanoparticle-based TR-FRET. Energy transfer between the solid-phase donors and Cy5-labeled protein were obtained with signal-to-background ratios ranging from 1.2 to 9.9. In this study, a thin layer prepared using Eu(III)-labeled protein gave the most efficient TR-FRET. This thin donor layer was tested in a competitive separation-free immunoassay of human albumin (hAlb). hAlb was measured in a clinically relevant concentrations from 0.05 to 10 mg l~(-1) with the coefficient of variation ranging from 1.0% to 12.4%.
机译：镧系元素螯合物和镧系元素纳米颗粒标记物是用于无分离时间分辨荧光共振能量转移（TR-FRET）分析的有吸引力的供体。在完全染色的纳米颗粒中，TR-FRET分析中纳米颗粒标记的内部体积由于与表面受体的距离过长而无法参与能量转移。我们的兴趣是研究基于表面的TR-FRET，因此，研究了各种（（III）（Eu）螯合物层的TR-FRET效率。制备掺入硅氧烷层中的Eu（III）螯合物，共价偶联于硅烷化表面和Eu（III）标记的蛋白质表面的Eu（III）螯合物，并将其与基于纳米颗粒的TR-FRET进行比较。固相供体与Cy5标记的蛋白质之间的能量转移以1.2至9.9的信号背景比获得。在这项研究中，使用Eu（III）标记的蛋白制备的薄层给出了最有效的TR-FRET。该薄供体层在人白蛋白（hAlb）的无竞争竞争性免疫测定中进行了测试。 hAlb的临床相关浓度为0.05至10 mg l〜（-1），变异系数为1.0％至12.4％。

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  • 来源
    《Applied Surface Science》 |2009年第14期|6529-6534|共6页
  • 作者

    Harri Haermae; Riikka Suhonen; Terho Kololuoma; Ari Kaerkkaeinen; Mika Hara; Pekka Haenninen;

  • 作者单位

    Laboratory of Biophysics, University of Turku, Tykistoekatu 6A, FIN-20520, Turku, Finland Department of Biotechnology, University of Turku, Tykistoekatu 6A, FIN-20520, Turku, Finland;

    VTT, Kaitovaeylae 1, FIN-90570, Oulu, Finland;

    VTT, Kaitovaeylae 1, FIN-90570, Oulu, Finland;

    BraggOne Oy, Kaitovaeylae 1, F1N-90570, Oulu, Finland;

    Laboratory of Biophysics, University of Turku, Tykistoekatu 6A, FIN-20520, Turku, Finland;

    Laboratory of Biophysics, University of Turku, Tykistoekatu 6A, FIN-20520, Turku, Finland;

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  • 原文格式 PDF
  • 正文语种 eng
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  • 关键词

    TR-FRET; europium; thin solid surface; immunoassay;

    机译：TR-FRET;铕;薄的固体表面;免疫测定;

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