开发了一种用于检测程序死亡配体 1 (PD-L1) 的一次性双输出生物传感器，用于在单一实验设置中监测免疫治疗进展和早期癌症检测。将适配体探针组装在与羧化三噻吩聚合物复合的rGO（rGO-pTBA）上，特异性捕获用新型氧化还原介质邻氨基苯酚对磺酸标记的PD-L1蛋白，用于电流检测。通过电化学和表面分析实验对每个传感层进行表征，然后确认其传感性能。标准 PD-L1 蛋白检测的校准图显示了 0.5-100.0 pM 和 100.0-500.0 pM 两个动态范围，其中电流分析法的检测限为 0.20 ± 0.001 pM (RSD ≤5.2%)。通过使用两种检测方法检测 A549 肺癌细胞分泌的 PD-L1 和可溶性 PD-L1 (sPD-L1) 的临床相关血清水平来评估传感器的可靠性。此外，还通过量化一小群肺癌患者的 sPD-L1 水平来研究治疗试验。与健康个体 (16.2-19.6 pM) 相比，患者 (221.6-240.4 pM) 的 sPD-L1 水平显着更高。免疫治疗后，患者的PD-L1水平下降至126.7-141.2 pM的范围。结果表明，使用两种建议的方法成功地完成了治疗监测。此外，根据对免疫检查点相关蛋白的比较研究，PD-L1 是比颗粒酶 B 和干扰素-γ 更有效的生物标志物。版权所有 © 2024 Elsevier B.V。保留所有权利。

A disposable dual-output biosensor to detect program death-ligand 1 (PD-L1) was developed for immunotherapy progress monitoring and early cancer detection in a single experimental setup. The aptamer probe was assembled on rGO composited with carboxylated terthiophene polymer (rGO-pTBA) to specifically capture PD-L1 protein labeled with a new redox mediator, ortho-amino phenol para sulphonic acid, for amperometric detection. Each sensing layer was characterized through electrochemical and surface analysis experiments, then confirmed the sensing performance. The calibration plots for the standard PD-L1 protein detection revealed two dynamic ranges of 0.5-100.0 pM and 100.0-500.0 pM, where the detection limit was 0.20 ± 0.001 pM (RSD ≤5.2%) by amperometry. The sensor reliability was evaluated by detecting A549 lung cancer cell-secreted PD-L1 and clinically relevant serum levels of soluble PD-L1 (sPD-L1) using both detection methods. In addition, therapeutic trials were studied through the quantification of sPD-L1 levels for a small cohort of lung cancer patients. A significantly higher level of sPD-L1 was observed for patients (221.6-240.4 pM) compared to healthy individuals (16.2-19.6 pM). After immunotherapy, the patients' PD-L1 level decreased to the range of 126.7-141.2 pM. The results indicated that therapy monitoring was successfully done using both the proposed methods. Additionally, based on a comparative study on immune checkpoint-related proteins, PD-L1 is a more effective biomarker than granzyme B and interferon-gamma.Copyright © 2024 Elsevier B.V. All rights reserved.