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首页> 外文会议>Conference on reporters, markers, dyes, nanoparticles, and molecular probes for biomedical applications >Activatable optical imaging probes with various fluorophore-quencher combinations
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Activatable optical imaging probes with various fluorophore-quencher combinations

机译：具有各种荧光团猝灭剂组合的可激活光学成像探针

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Molecular imaging probes rely on high target-to-background ratios (TBR) to achieve maximum sensitivity and specificity. We utilized "quenchers" to turn off the background signal from the unbound probe and investigated the ability of specific fluorophore-quencher pairs to activate at target tissues. Both fluorophore and quencher were conjugated to a single cancer targeting molecule, either avidin or antibody. Fluorescence signal from these targeting molecules was "turned off' by the quencher in the unbound state, but was "turned on" only when the molecules bound to the cell surface target and was internalized. We tested the following fluorophore-quencher combinations based on fluorescence resonance energy transfer (FRET) pairs; OregonG-BHQ1, RhodG-BHQ1/ATT0540Q, TAMRA-QSY7/QSY21, TexRed-QSY21, Alexa647-QSY21, Cy5.5-QSY21/BHQ3 and Alexa680-QSY21/BHQ3. Among these, only RhodGATTO540Q and TAMRA-QSY7/21 pair showed activation upon cell binding/internalization. Among these combinations, TAMRA-QSY7 pair showed the highest activation (40-fold and 13-fold for avidin and antibody conjugate, respectively) as measured with an in vitro dissociation assay. The activation was dependent on the method used to conjugate fluorophores and quenchers to the targeting molecule. In vitro microscopic studies with TAMRA-QSY7 pair conjugated to avidin or antibody showed high fluorescent signal inside the target cancer cells, indicating activation after internalization. In vivo imaging studies in tumor bearing mice demonstrated that tumors could be clearly detected with low background. Although the precise quenching mechanism remains to be determined, this activation system can achieve high TBR in vivo molecular imaging.
机译：分子成像探针依赖于高靶标比率（TBR）以实现最大敏感性和特异性。我们利用“猝灭剂”从未结合探针关闭背景信号，并研究了特定荧光团猝灭剂对在靶组织中激活的能力。荧光团和猝灭剂均与单一癌症靶向分子缀合物，抗生物素蛋白或抗体。来自这些靶向分子的荧光信号被猝灭剂在未结合状态下“关闭”，但是仅在与细胞表面靶标结合并内化的分子时“打开”。我们基于荧光测试以下荧光团猝灭剂组合共振能量转移（FRET）对; OREGONG-BHQ1，RHODG-BHQ1 / ATT0540Q，TAMRA-QSY7 / QSY21，TEXRED-QSY21，ALEXA647-QSY21，CY5.5-QSY21 / BHQ3和ALEXA680-QSY21 / BHQ3。其中只有RhodGATTO540Q和TAMRA-QSY7 / 21对显示在细胞结合/内化活化。在这些组合中，TAMRA-QSY7对显示最高的活化（40倍和，分别为13倍为抗生物素蛋白和抗体缀合物），其具有在体外测量解离测定。活化取决于用于将荧光团和猝灭剂缀合到靶向分子的方法。与抗氨蛋白或抗体缀合的TAMRA-QSY7对的体外显微研究显示靶癌中的高荧光信号ER细胞，在内化后表明激活。在肿瘤轴承小鼠的体内成像研究中表明，可以用低背景清楚地检测到肿瘤。尽管仍有待确定的精确淬火机制，但这种活化系统可以在体内分子成像中实现高TBR。

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