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首页> 外文会议>Conference on Optical Diagnostics of Living Cells Ⅴ, Jan 23-25, 2002, San Jose, USA >Optimizing the Luminescence of Lanthanide(Ⅲ) Macrocyclic Complexes for the Detection of Anti5BrdU
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Optimizing the Luminescence of Lanthanide(Ⅲ) Macrocyclic Complexes for the Detection of Anti5BrdU

机译：镧系（Ⅲ）大环配合物的发光优化用于检测抗5BrdU

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摘要

A Eu(Ⅲ)-macrocycle-mono-isothiocyanate, Quantum Dye~(~R), has been coupled to a monoclonal antibody against 5BrdU. Since Quantum Dyes do not undergo concentration quenching, the coupling conditions were optimized to achieve the maximum number of Eu(Ⅲ) macrocyles bound to the antiBrdU, without decrease in solubility or loss of antigen-binding ability. In order to optimize the coupling conditions, a colorimetric method for the quantitation of the Eu(Ⅲ)-macrocycle-mono-isothio-cyanate has been developed. A simple mixture composed of an ethanolic solution and a Gd(Ⅲ)-containing aqueous solution is now used to provide lan-thanide enhanced luminescence, LEL. Under LEL conditions, the specific binding of Eu(Ⅲ) macrocycles to apoptotic cells has been observed in both aqueous and mounted slide preparations. A comparison between measurements of the same LEL model system, obtained in both time-gated luminescence and standard fluorescence modes, has demonstrated that time-gating significantly improves the signal to noise ratio.
机译：Eu（Ⅲ）-大环-单-异硫氰酸酯，Quantum Dye_（R）已与抗5BrdU的单克隆抗体偶联。由于量子染料不进行浓度猝灭，因此优化了偶联条件，以实现与抗BrdU结合的Eu（Ⅲ）巨环的最大数量，而不会降低溶解度或抗原结合能力。为了优化偶联条件，开发了比色法定量Eu（Ⅲ）-大环-单-异硫氰酸酯。现在，由乙醇溶液和含Gd（Ⅲ）的水溶液组成的简单混合物可提供镧系元素增强的发光LEL。在LEL条件下，在水性和固定载玻片制剂中均观察到Eu（Ⅲ）大环与凋亡细胞的特异性结合。在时间门控发光和标准荧光模式下获得的相同LEL模型系统的测量结果之间的比较表明，时间门控可显着提高信噪比。

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