Optical assay for glucose based on the luminescnence decay time of the long wavelength dye Cy5™.
Optical assay for glucose based on the luminescnence decay time of the long wavelength dye Cy5™.
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基于长波长染料 Cy5™ 的发光衰减时间的葡萄糖光学测定。
DOI：
10.1016/s0925-4005(97)00275-x
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Lakowicz,JosephR
中科院分区：
文献类型：
--
作者：
Tolosa,Leah;Malak,Henryk;Raob,Govind;Lakowicz,JosephR
基于长波长染料 (Cy5) 的发光衰减时间描述了葡萄糖的光学测定，该染料可以用当前可用的红色激光二极管激发。伴刀豆球蛋白 A 与 Cy5 共价标记，Cy5 在基于荧光共振能量转移 (FRET) 的测定中充当供体。受体是孔雀石绿，它与作为载体蛋白的胰岛素共价连接。为了提供 ConA 孔雀绿胰岛素的结合亲和力，还用麦芽糖 (MIMG) 共价标记胰岛素。通过时间相关的单光子计数观察到，Cy5ConA 与 MIMG 的结合导致 Cy5 的强度和衰减时间降低。通过从 Cy5ConA 竞争性置换 MIMG 来检测葡萄糖，从而导致强度和衰减时间增加。这种葡萄糖测定有几个特点，可以实现现实世界中葡萄糖的实际测定。 Cy5 的长吸收波长允许用红色激光二极管进行激发，可以轻松地对红色激光二极管进行脉冲或幅度调制，以进行时域或频域衰减时间测量。此外，可以使用长波长激发和发射通过皮肤测量衰减时间，这表明植入葡萄糖传感器的可能性。最后，原则上可以通过控制载体蛋白糖标记的程度和类型来调整测定亲和力和可逆性。
An optical assay for glucose is described based on the luminescence decay time of a long wavelength dye (Cy5) which can be excited with currently available red laser diodes. Concanavalin A was covalently labeled with Cy5 which served as the donor in an assay based on fluorescence resonance energy transfer (FRET). The acceptor was Malachite Green which was covalently linked to insulin which served as a carrier protein. To provide binding affinity for ConA Malachite Green insulin was also covalently labeled with maltose (MIMG). Binding of Cy5ConA to MIMG resulted in a decreased intensity and decay time of Cy5 as observed by time-correlated single photon counting. Glucose was detected by competitive displacement of MIMG from Cy5ConA, resulting in increased intensity and decay time. This glucose assay has several features which can result in practical real world assays for glucose. The long absorption wavelength of Cy5 allows excitation with red laser diodes, which can be readily pulsed or amplitude-modulated for time-domain or frequency-domain decay time measurements. Additionally, decay times can be measured through skin using long wavelength excitation and emission, suggesting the possibility of an implanted glucose sensor. And finally, the assay affinity and reversibility can in principle be adjusted by controlling the extent and type of sugar labeling of the carrier protein.
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