School of Chemistry and Chemical Engineering, Shandong University, Jinan 250100, China.
Shenzhen Lifotronic Technology Co., Ltd, Nanshan District, Shenzhen 518055, China.
Anal Chem. 2022 Nov 15;94(45):15801-15808. doi: 10.1021/acs.analchem.2c03579. Epub 2022 Nov 5.
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 Zn-mediated aggregation-induced emission (AIE) assembly of Au nanocrystals (NCs) (Zn-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 Zn-AIE-AuNCs can multiplex with the single-waveband and surface-defect-involved ECL (775 nm) of dual-stabilizer-capped CuInS@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 Zn-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.
电化学发光（ECL）多重生物分析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 多重生物分析中短波段的限制，本文开发了一种高单色性（半峰全宽<40nm）和能带工程化的 ECL 发光体，即巯基丙酸封端和 Zn 介导的聚集诱导发射（AIE）组装的 Au 纳米晶体（NCs）（Zn-AIE-AuNCs），其发光强度高，最大发射波长为 485nm。Zn-AIE-AuNCs 的高单色性和能带工程化 ECL（485nm）可以与双稳定剂封端的 CuInS@ZnS NCs（CIS@ZnS-NCs）的单波段和表面缺陷相关的 ECL（775nm）进行多重化，实现了具有相似颗粒大小的不同 NCs 标记物的光谱分辨 ECL 多重化策略。该 ECL 多重化策略可用于同时检测抗原和 DNA 探针，而无需任何额外的信号放大步骤和明显的光谱串扰，其中 Zn-AIE-AuNCs 的高单色性 ECL 用于动态检测人癌胚抗原，从 1pg/mL 到 50ng/mL，检测限（LOD）为 0.3pg/mL，而 CIS@ZnS-NCs 的单波段 ECL 用于线性检测野生型 p53，从 1pM 到 50nM，LOD 为 0.5pM。所提出策略的 ECL 免疫分析不受同步进行的 DNA 探针分析的干扰，反之亦然，这将为将免疫分析和 DNA 探针分析结合在一起用于临床结肠癌和乳腺癌识别开辟一条途径。
