School of Pharmaceutical Science, Xiamen University, 4221 Xiang'an South Road, Xiamen 361102, China.
Department of Cardiac Surgery, Xiamen Cardiovascular Hospital, Xiamen University, 2999 Jinshan Road, Xiamen 361010, China.
Biosensors (Basel). 2023 Jun 13;13(6):647. doi: 10.3390/bios13060647.
In this study, a label-free and antibody-free impedimetric biosensor based on molecularly imprinting technology for exosomes derived from non-small-cell lung cancer (NSCLC) cells was established. Involved preparation parameters were systematically investigated. In this design, with template exosomes anchored on a glassy carbon electrode (GCE) by decorated cholesterol molecules, the subsequent electro-polymerization of APBA and elution procedure afforded a selective adsorption membrane for template A549 exosomes. The adsorption of exosomes caused a rise in the impedance of the sensor, so the concentration of template exosomes can be quantified by monitoring the impedance of GCEs. Each procedure in the establishment of the sensor was monitored with a corresponding method. Methodological verification showed great sensitivity and selectivity of this method with an LOD = 2.03 × 10 and an LOQ = 4.10 × 10 particles/mL. By introducing normal cells and other cancer cells derived exosomes as interference, high selectivity was proved. Accuracy and precision were measured, with an obtained average recovery ratio of 100.76% and a resulting RSD of 1.86%. Additionally, the sensors' performance was retained at 4 °C for a week or after undergoing elution and re-adsorption cycles seven times. In summary, the sensor is competitive for clinical translational application and improving the prognosis and survival for NSCLC patients.
在这项研究中,建立了一种基于分子印迹技术的无标记和无抗体的阻抗生物传感器,用于检测非小细胞肺癌(NSCLC)细胞来源的外泌体。系统地研究了涉及的制备参数。在该设计中,模板外泌体通过修饰的胆固醇分子锚定在玻碳电极(GCE)上,随后进行 APBA 的电聚合和洗脱过程,为模板 A549 外泌体提供了选择性吸附膜。外泌体的吸附会导致传感器阻抗升高,因此可以通过监测 GCE 的阻抗来定量模板外泌体的浓度。传感器建立的每个步骤都使用相应的方法进行监测。方法学验证表明,该方法具有很高的灵敏度和选择性,LOD = 2.03×10,LOQ = 4.10×10 个/mL。通过引入正常细胞和其他癌细胞来源的外泌体作为干扰,证明了该方法具有很高的选择性。对准确性和精密度进行了测量,得到的平均回收率为 100.76%,RSD 为 1.86%。此外,传感器的性能在 4°C 下可保持一周,或经过洗脱和再吸附循环七次后仍可保持。总之,该传感器具有竞争力,可用于临床转化应用,改善 NSCLC 患者的预后和生存率。