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Spectrum-Resolved Electrochemiluminescence to Multiplex the Immunoassay and DNA Probe Assay
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-11-05 , DOI: 10.1021/acs.analchem.2c03579
Xuwen Gao 1 , Xiancheng Liu 2 , Ying Zeng 2 , Qingqing Zhang 2 , Bin Zhang 1 , Guizheng Zou 1
Affiliation  

The investigation on electrochemiluminescence (ECL) multiplexing bioassays mainly focuses on simultaneously detecting either proteins or nucleic acids. To overcome the limitation of a short waveband for spectrum-resolved ECL multiplexing bioassays, herein, a highly monochromatic (FWHM <40 nm) and bandgap-engineered ECL luminophore, that is, mercaptopropionic acid-capped and Zn2+-mediated aggregation-induced emission (AIE) assembly of Au nanocrystals (NCs) (Zn2+-AIE-AuNCs), of strong emission and the maximum emission wavelength at 485 nm is developed. The highly monochromatic and bandgap-engineered ECL (485 nm) of Zn2+-AIE-AuNCs can multiplex with the single-waveband and surface-defect-involved ECL (775 nm) of dual-stabilizer-capped CuInS2@ZnS NCs (CIS@ZnS-NCs), enabling a spectrum-resolved ECL multiplexing strategy with different NCs luminophores of a similar particle size as tags. This ECL multiplexing strategy can be utilized to simultaneously detect antigen and DNA probe together without any additional signal amplification procedure and obvious spectroscopic cross-talk, in which the highly monochromatic ECL from Zn2+-AIE-AuNCs is utilized to dynamically determine human carcinoembryonic antigen from 1 pg/mL to 50 ng/mL with a limit of detection (LOD) of 0.3 pg/mL, while the single-waveband ECL from CIS@ZnS-NCs is employed to linearly detect wild-type p53 from 1 pM to 50 nM with a LOD of 0.5 pM. The ECL immunoassay of the proposed strategy is free from the interference of the synchronously conducted DNA probe assay and vice versa, which would open an avenue to couple the immunoassay and DNA probe assay together for clinical colon and breast cancer identification.

中文翻译:

光谱分辨电化学发光复用免疫测定和 DNA 探针测定

电化学发光 (ECL) 多重生物测定的研究主要集中在同时检测蛋白质或核酸。为了克服光谱分辨 ECL 多重生物测定的短波段限制,本文采用高度单色(FWHM <40 nm)和带隙工程化的 ECL 发光体,即巯基丙酸封端和 Zn 2+ 介导的聚集诱导开发了 Au 纳米晶体 (NCs) (Zn 2+ -AIE-AuNCs)的发射 (AIE) 组件,具有强发射和 485 nm 的最大发射波长。Zn 2+ -AIE-AuNCs 的高度单色和带隙设计的 ECL (485 nm)可以与双稳定剂封端的 CuInS 2的单波段和涉及表面缺陷的 ECL (775 nm) 复用@ZnS NCs (CIS@ZnS-NCs),使用与标签具有相似粒径的不同 NCs 发光团,实现光谱分辨 ECL 复用策略。这种 ECL 复用策略可用于同时检测抗原和 DNA 探针,无需任何额外的信号放大程序和明显的光谱串扰,其中来自 Zn 2+ 的高度单色ECL-AIE-AuNCs用于动态测定1 pg/mL至50 ng/mL的人癌胚抗原,检测限(LOD)为0.3 pg/mL,同时采用CIS@ZnS-NCs的单波段ECL线性检测 1 pM 至 50 nM 的野生型 p53,LOD 为 0.5 pM。所提出策略的 ECL 免疫测定不受同步进行的 DNA 探针测定的干扰,反之亦然,这将为将免疫测定和 DNA 探针测定结合在一起用于临床结肠癌和乳腺癌鉴定开辟一条途径。
更新日期:2022-11-05
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