From antenna to assay: lessons learned in lanthanide luminescence.
From antenna to assay: lessons learned in lanthanide luminescence.
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DOI：
10.1021/ar800211j
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发表时间：
2009-04-21
影响因子：
18.3
通讯作者：
Raymond, Kenneth N.
中科院分区：
文献类型：
--
作者：
Moore, Evan G.;Samuel, Amanda P. S.;Raymond, Kenneth N.
配体敏化的发光镧系元素（III）络合物具有相当重要的意义，因为它们独特的生物物理性质（微秒至毫秒的寿命、特征和窄的发射带以及大的斯托克斯位移）使它们非常适合作为基于荧光的生物测定中的标记物。镧系元素（III）阳离子的长寿命发射可以从散射光和背景荧光中暂时分辨出来，以大大提高测量灵敏度。该领域的一个挑战是敏化配体的设计，其提供具有足够稳定性和水溶性的高发射性络合物用于实际应用。在这个帐户中，我们给出了一些三价镧系元素的一般性质的概述，并随后在我们的实验室在生物技术中的应用程序的高度发光的Tb（III）和Eu（III）配合物的开发所取得的进展的总结。我们的研究重点是优化这些化合物作为用于均相时间分辨荧光（HTRF）技术的潜在商业试剂。我们的方法涉及开发高稳定性的八齿Tb（III）和Eu（III）络合物，其依赖于全氧供体原子，并使用多发色团螯合物来增加摩尔吸光率;早期的例子利用了单个悬垂发色团（即，单个“天线”）。基于2-羟基乙酰胺（IAM）的配体提供了非常发光的Tb（III）配合物，其量子产率值高达60%，在商业测定所需的纳摩尔浓度下是稳定的。通过合成修饰的IAM生色团和含时密度泛函理论（TD-DFT）计算，我们已经开发了一种方法来预测这些生色团的吸收和发射性质作为一种工具，以指导配体设计。此外，我们还研究了手性IAM配体，产生Tb（III）配合物具有高的量子产率值和强的圆偏振发光（CPL）的活动。为了有效地敏化Eu（III）发射，我们已经使用1-羟基吡啶-2-酮（1，2-HOPO）螯合物来产生显著的配体，其结合了优异的物理性质和优异的水稳定性的联合收割机。通过将低温磷光测量与IAM系统使用的相同TD-DFT方法相结合，已经实现了对这种发色团的更完整的理解。Eu（III）与手性1，2-HOPO配体的配合物也具有很强的CPL活性。我们还进行了一系列Eu（III）配合物的辐射和非辐射衰变途径的动力学分析;金属离子对称性对随后的物理性质的重要性是清楚的。最后，我们描述了一个Tb（III）-IAM化合物，现在进行到商业可用性，提供了改进的性能，在共同的HTRF平台，并有可能大大提高灵敏度。
Ligand-sensitized, luminescent lanthanide(III) complexes are of considerable importance because their unique photophysical properties (microsecond to millisecond lifetimes, characteristic and narrow emission bands, and large Stokes shifts) make them well suited as labels in fluorescence-based bioassays. The long-lived emission of lanthanide(III) cations can be temporally resolved from scattered light and background fluorescence to vastly enhance measurement sensitivity. One challenge in this field is the design of sensitizing ligands that provide highly emissive complexes with sufficient stability and aqueous solubility for practical applications. In this Account, we give an overview of some of the general properties of the trivalent lanthanides and follow with a summary of advances made in our laboratory in the development of highly luminescent Tb(III) and Eu(III) complexes for applications in biotechnology. A focus of our research has been the optimization of these compounds as potential commercial agents for use in Homogeneous Time-Resolved Fluorescence (HTRF) technology. Our approach involves developing high-stability octadentate Tb(III) and Eu(III) complexes that rely on all-oxygen donor atoms and using multi-chromophore chelates to increase molar absorptivity; earlier examples utilized a single pendant chromophore (that is, a single “antenna”). Ligands based on 2-hydroxyisophthalamide (IAM) provide exceptionally emissive Tb(III) complexes with quantum yield values up to ∼60% that are stable at the nanomolar concentrations required for commercial assays. Through synthetic modification of the IAM chromophore and time-dependent density functional theory (TD-DFT) calculations, we have developed a method to predict absorption and emission properties of these chromophores as a tool to guide ligand design. Additionally, we have investigated chiral IAM ligands that yield Tb(III) complexes possessing both high quantum yield values and strong circularly polarized luminescence (CPL) activity. To efficiently sensitize Eu(III) emission, we have used the 1-hydroxypyridin-2-one (1,2-HOPO) chelate to create remarkable ligands that combine excellent photophysical properties and exceptional aqueous stabilities. A more complete understanding of this chromophore has been achieved by combining low-temperature phosphorescence measurements with the same TD-DFT approach used with the IAM system. Eu(III) complexes with strong CPL activity have also been obtained with chiral 1,2-HOPO ligands. We have also undertaken the kinetic analysis of radiative and non-radiative decay pathways for a series of Eu(III) complexes; the importance of the metal ion symmetry on the ensuing photophysical properties is clear. Lastly, we describe a Tb(III)-IAM compound—now carried through to commercial availability—that offers improved performance in the common HTRF platform and has the potential to vastly improve sensitivity.
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影响因子：
15
作者：
Seitz, Michael;Moore, Evan G.;Raymond, Kenneth N.
通讯作者：
Raymond, Kenneth N.
影响因子：
3.6
作者：
Latva, M;Takalo, H;Kankare, J
通讯作者：
Kankare, J
影响因子：
4.6
作者：
Samuel, Amanda P. S.;Xu, Jide;Raymond, Kenneth N.
通讯作者：
Raymond, Kenneth N.
影响因子：
2.9
作者：
HEMMILA, I;DAKUBU, S;LOVGREN, T
通讯作者：
LOVGREN, T
影响因子：
4.6
作者：
Samuel AP;Moore EG;Melchior M;Xu J;Raymond KN
通讯作者：
Raymond KN