Li Xiwen, Meng Peiran, Sun Mingyang, Chen Yue, Song Zhiyi, Wang Xinyao, Li Na, Sun Yue
School of Chemistry and Chemical Engineering, Liaoning Normal University, Dalian 116029, China.
Analyst. 2023 May 16;148(10):2352-2361. doi: 10.1039/d3an00234a.
Phospholipase C (PLC) has important biological functions in specific cancer types, immune disorders and neurodegeneration. Here, an ultrasensitive electrochemical sensor for PLC was developed signal amplification based on breathing atom transfer radical polymerization (ATRP). First, phosphatidylethanolamine (PE) was immobilized on the surface of a gold electrode by L-cysteine and cross-linker. Then, PE was specially hydrolyzed by PLC to obtain the phosphate groups and tethered with the ATRP initiator α-bromophenacetic acid (BPAA) by the coordination action of Zr. After the breathing ATRP, a large number of electroactive monomers (ferrocenylmethyl methacrylate, FcMMA) were successfully grafted from BPAA. The experimental results indicated that the detection signal of the obtained electrode (sensor) was proportional to the concentration of PLC. The sensor showed a low detection limit of 0.270 U L and a wide linear range of 1-40 U L ( = 0.997). Most importantly, the sensor was successfully applied to detect PLC in breast cancer cells (MCF-7, MDA-MB-231) and nontumor cells (MCF-10A). The value obtained by our electrochemical sensor had no obvious difference from that determined by the commercial ELISA kit. These results showed that the fabricated PLC sensor had acceptable potential in clinical applications.
磷脂酶C（PLC）在特定癌症类型、免疫紊乱和神经退行性变中具有重要的生物学功能。在此，基于呼吸原子转移自由基聚合（ATRP）信号放大技术开发了一种用于检测PLC的超灵敏电化学传感器。首先，通过L-半胱氨酸和交联剂将磷脂酰乙醇胺（PE）固定在金电极表面。然后，PE被PLC特异性水解以获得磷酸基团，并通过Zr的配位作用与ATRP引发剂α-溴苯乙酸（BPAA）相连。经过呼吸ATRP后，大量的电活性单体（甲基丙烯酸二茂铁甲酯，FcMMA）成功地从BPAA接枝。实验结果表明，所得电极（传感器）的检测信号与PLC的浓度成正比。该传感器的检测限低至0.270 U/L，线性范围宽达1 - 40 U/L（R² = 0.997）。最重要的是，该传感器成功应用于检测乳腺癌细胞（MCF - 7、MDA - MB - 231）和非肿瘤细胞（MCF - 10A）中的PLC。我们的电化学传感器所获得的值与商业ELISA试剂盒测定的值没有明显差异。这些结果表明，所制备的PLC传感器在临床应用中具有可接受的潜力。
