School of Chemistry and Chemical Engineering , Shandong University , Jinan , 250100 , China.
Anal Chem. 2018 Apr 17;90(8):5474-5480. doi: 10.1021/acs.analchem.8b00831. Epub 2018 Apr 6.
The electrochemical-dependent and unstable intensity of electrochemiluminescence (ECL) makes it difficult to identify ECL in a waveband-resolved way on photomultiplier tube (PMT)-based traditional ECL analyzers. Herein, a dual-color ECL strategy is proposed by transferring ECL to two PMTs in a waveband-resolved way via dichroic mirror, simultaneously detecting wild-type p53 (WTp53) in near-infrared wavebands with CdTe (λ = 782 nm) nanocrystals as tag and mutant p53 (MUp53) in eye-visible wavebands with CdSe (λ = 554 nm) nanocrystals as tag. The two targets can be color-selectively determined from 10 fM to 100 pM with a limit of detection at 5 fM for MUp53 and from 50 fM to 100 pM with a limit of detection at 10 fM for WTp53, respectively. The dichroic mirror-involved ECL setup is easy to assemble for popularization, which would not only eventually enable PMT-based multiple-color ECL analysis but also make it is possible to directly determine the changed level of tumor suppressors for cancer diagnosis and therapeutic evaluation via one-pot ECL assay.
电化学发光（ECL）的电化学依赖性和不稳定性使其难以在基于光电倍增管（PMT）的传统 ECL 分析仪上以波段分辨的方式识别 ECL。在此，通过二向色镜将 ECL 以波段分辨的方式转移到两个 PMT 上，提出了双色 ECL 策略，同时使用碲化镉（CdTe，λ=782nm）纳米晶体作为标记物在近红外波段检测野生型 p53（WTp53），使用硒化镉（CdSe，λ=554nm）纳米晶体作为标记物在可见波段检测突变型 p53（MUp53）。这两个目标物可以从 10 fM 到 100 pM 进行颜色选择性测定，对于 MUp53 的检测限为 5 fM，对于 WTp53 的检测限为 10 fM 从 50 fM 到 100 pM。涉及二向色镜的 ECL 装置易于组装，便于推广，不仅最终能够实现基于 PMT 的多色 ECL 分析，而且还可以通过一次性 ECL 测定法直接确定肿瘤抑制剂的变化水平，用于癌症诊断和治疗评估。
