Key Laboratory for Biorheological Science and Technology of Ministry of Education, Bioengineering College of Chongqing University, Chongqing, 400044, PR, China.
Chongqing University Three Gorges Hospital, Chongqing, 404000, PR, China.
Biosens Bioelectron. 2022 Oct 15;214:114508. doi: 10.1016/j.bios.2022.114508. Epub 2022 Jun 24.
In vitro detection of biomarkers requires ultrahigh sensitivity and accuracy. The use of a dual-signal (electrochemical and fluorescent) strategy has the ability of self-calibration and can overcome interferences seen from experimental and environmental factors influencing the experimental results, showing significant advantages in the detection of biomarkers. Here, we propose an enzyme-free and label-free dual-signal aptasensor based on functionalized Apt@antimonene quantum dot (Sb)@ methylene blue (MB)@ZIF-67/Apt@Sb@3,3',5,5'-tetramethylbenzidine (TMB) @ZIF-67 material to simultaneously detect multiple tumour biomarkers -oestrogen receptor (ER) and human epidermal growth factor receptor-2 (HER2). The aptamer chain of HER2 and ER is adsorbed to cover the porous structure of ZIF-67 through Sb, which has the characteristics of single-chain adsorption, in which the target protein is desorbed later, further encapsulating its internal MB and TMB with good dual-signal properties. After Apt@Sb@MB@ZIF-67/Apt@Sb@TMB@ZIF-67 targeted HER2 and ER biomarkers, the aptamer chains on the Sb quantum dots were desorbed and transferred into the solution, and the MB and TMB signals were released. Negative pressure is applied to the ITO electrode to adsorb and enrich positively charged MB and TMB organic molecules for electrochemical detection, and the supernatant after centrifugation can be directly subjected to fluorescence detection. Therefore, accurate electrochemical and fluorescent dual-signal detection of HER2 and ER from tissue to serum is achieved. Notably, sensitive detection of HER2-ER biomarkers was achieved in only 30 min, with a detection range of 0-10 pg/mL and detection limits of 3.4 fg/mL and 6.9 fg/mL, respectively.
在体外检测生物标志物需要超高的灵敏度和准确性。采用双信号(电化学和荧光)策略具有自校准能力,可以克服实验和环境因素对实验结果的干扰,在生物标志物检测方面显示出显著的优势。在这里,我们提出了一种基于功能化 Apt@antimonene 量子点(Sb)@亚甲蓝(MB)@ZIF-67/Apt@Sb@3,3',5,5'-四甲基联苯胺(TMB)@ZIF-67 材料的无酶和无标记的双信号适体传感器,用于同时检测多种肿瘤生物标志物-雌激素受体(ER)和人表皮生长因子受体-2(HER2)。HER2 和 ER 的适体链通过 Sb 吸附覆盖 ZIF-67 的多孔结构,具有单链吸附的特点,随后目标蛋白被解吸,进一步用具有良好双信号特性的内部 MB 和 TMB 进行封装。在 Apt@Sb@MB@ZIF-67/Apt@Sb@TMB@ZIF-67 靶向 HER2 和 ER 生物标志物后,Sb 量子点上的适体链被解吸并转移到溶液中,释放出 MB 和 TMB 信号。在 ITO 电极上施加负压以吸附和富集带正电的 MB 和 TMB 有机分子进行电化学检测,离心后的上清液可直接进行荧光检测。因此,实现了对组织到血清中 HER2 和 ER 的准确电化学和荧光双信号检测。值得注意的是,仅用 30 分钟即可实现对 HER2-ER 生物标志物的灵敏检测,检测范围为 0-10 pg/mL,检测限分别为 3.4 fg/mL 和 6.9 fg/mL。