Hydrophilic labeling reagents of dipyrrylmethene-BF2 dyes for two-photon excited fluorometry:: Syntheses and photophysical characterization
Hydrophilic labeling reagents of dipyrrylmethene-BF2 dyes for two-photon excited fluorometry:: Syntheses and photophysical characterization
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DOI：
10.1023/b:jofl.0000039350.94256.53
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发表时间：
2004-09-01
影响因子：
2.7
通讯作者：
Soini, AE
中科院分区：
文献类型：
--
作者：
Meltola, NJ;Wahlroos, R;Soini, AE
最近推出的生物亲和分析技术ArcDia(TM)TPX是基于双光子激发荧光(TPE)的，它可以从微体积中进行免分离的超灵敏免疫分析。在这里，我们介绍了新型双光子可激发荧光标记试剂的合成，这些试剂是专门设计用于ArcDia(TM)TPX分析技术的标记分子。标记试剂是基于二吡咯甲基二氟化硼(DPMA-BF2)的发色团，这些发色团被芳基、杂芳基或芳基烯基取代，以扩大pi-电子共轭。这些取代反应得到了一系列具有不同光物理性质的二吡咯亚甲基-BF2荧光团。二吡咯亚甲基-BF2荧光团进一步被二肽连接单元取代，并最终以琥珀酰亚胺酯的形式被激活，以实现与伯氨基的特定偶联。二肽连接物充当标记和靶之间的间隔臂，并提高标记在水溶液中的溶解度。对新型标记试剂的化学和光物理性能进行了研究。新型标记试剂具有较高的荧光量子产率和摩尔吸收系数。结果表明，含亲水二肽连接单元的新型标记物具有较大的双光子激发截面、较高的荧光量子效率和较好的水溶液溶解性。结果表明，新型双吡咯亚甲基-BF2标记非常适用于基于双光子激发荧光的生物亲和分析。
Recently introduced bioaffinity assay technology, ArcDia(TM)TPX, is based on two-photon excited fluorescence (TPE) and it enables separation-free ultra-sensitive immunoassays from microvolumes. Here we present syntheses of novel two-photon excitable fluorescent labeling reagents which have been specially designed to be used as label molecules in the ArcDia(TM)TPX assay technique. The labeling reagents are based on dipyrrylmetheneboron difluoride (dipyrrylmethene-BF2) chromophore, which have been substituted with aryl, heteroaryl or arylalkenyl chemical groups to extend the pi-electron conjugation. These substitutions results in a series of dipyrrylmethene-BF2 fluorophores with different photophysical properties. Dipyrrylmethene-BF2 fluorophores have been further substituted with a dipeptide linker unit and finally activated as succinimidyl esters to enable specific coupling with primary amino groups. The dipeptide linker serves as a spacer arm between the label and a target, and enhances the solubility of the label in aqueous solutions. Study of the chemical and photophysical performance of the new labeling reagents is described. The new labeling reagents exhibit high fluorescence quantum yields, and molar absorption coefficients. The results show that the new labels with the hydrophilic dipeptide linker unit provide large two-photon excitation cross-sections, high fluorescence quantum efficiency and good solubility in aqueous solutions. The results suggest that the novel dipyrrylmethene-BF2 labels are highly applicable to bioaffinity assays based on two-photon excitation of fluorescence.