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Electrochemiluminescent detection of epilepsy biomarker miR-134 using a metal complex light switch
Bioelectrochemistry ( IF 5 ) Pub Date : 2022-05-05 , DOI: 10.1016/j.bioelechem.2022.108150
Robert J Forster 1 , David C Henshall 2 , Hany El Naggar 3 , Yann Pellegrin 4 , Norman Delanty 3
Affiliation  

The detection of a key biomarker in epilepsy, miR-134, using an environmentally sensitive electrochemiluminescent luminophore, [Ru(DPPZ)2 PIC]2+, is reported, DPPZ is dipyrido[3,2-a:2′,3′-c]phenazine) and PIC is (2,2′-bipyridyl)-2(4-carboxy phenyl) imidazo [4,5][1,10] phenanthroline. A thiolated capture strand is first labelled with [Ru(DPPZ)2 PIC]2+ and then adsorbed onto a gold electrode. No significant electrochemiluminescence, ECL, is observed for immobilised Ru-labelled capture strands which is consistent with the light-switch dye being exposed to the aqueous solution. In sharp contrast, binding of the target turns on ECL. The ECL intensity, IECL, depends on the number of adenine “spacer” bases between the end of the capture sequence and the dye. The ECL intensity for the optimised system increases linearly with increasing miR-134 concentration from 100 nM to approximately 20 μM. Single and double base mismatches produce IECL that are only approximately 30% and 8% respectively of that observed for the fully complementary target reflecting differences in their association constants. Significantly, the presence of BSA protein causes IECL to increase by less 5% in either the single or duplex circumstances. Finally, the ability of the sensor to quantify miR-134 in unprocessed plasma samples from healthy volunteers and people with epilepsy is reported.



中文翻译:

使用金属络合物光开关电化学发光检测癫痫生物标志物 miR-134

据报道,使用环境敏感的电化学发光体 [Ru(DPPZ) 2 PIC] 2+检测癫痫中的关键生物标志物 miR-134 ,DPPZ 是双吡啶并[3,2-a:2',3'- c]吩嗪)和PIC是(2,2'-联吡啶)-2(4-羧基苯基)咪唑并[4,5][1,10]菲咯啉。首先用[Ru(DPPZ) 2 PIC] 2+标记硫醇化捕获链,然后吸附到金电极上。对于固定的钌标记的捕获链,没有观察到显着的电化学发光 ECL,这与暴露于水溶液中的光开关染料一致。形成鲜明对比的是,目标的结合开启了 ECL。ECL 强度,I ECL,取决于捕获序列末端和染料之间的腺嘌呤“间隔”碱基的数量。优化系统的 ECL 强度随着 miR-134 浓度从 100 nM 增加到大约 20 μM 线性增加。单碱基和双碱基错配产生的 I ECL分别仅为完全互补目标观察到的 ECL 的大约 30% 和 8%,反映了它们的关联常数的差异。重要的是,BSA 蛋白的存在导致 I ECL在单链或双链情况下增加不到 5%。最后,报告了传感器在来自健康志愿者和癫痫患者的未处理血浆样本中量化 miR-134 的能力。

更新日期:2022-05-10
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