双重信号输出生物传感器检测程序死亡配体1和癌症的治疗进度监测
Dual-signal output biosensor for the detection of program death-ligand 1 and therapy progress monitoring of cancer
影响因子:10.50000
分区:化学1区 Top / 生物物理1区 生物工程与应用微生物1区 分析化学1区 电化学2区
发表日期:2024 Oct 15
作者:
Heru Agung Saputra, Jae Heun Chung, Md Ali Zaber Sahin, Deog-Su Park, Yoon-Bo Shim
摘要
开发了一次性双输出生物传感器，以检测程序死亡 - 配体1（PD-L1）进行免疫疗法进度监测和单个实验设置中的早期癌症检测。将适体探针组装在与羧化的Terthiophene聚合物（RGO-PTBA）合成的RGO上，以专门捕获用新的氧化还原介质介质，Ortho-Amino para磺酸标记的PD-L1蛋白，以进行amperometric检测。通过电化学和表面分析实验来表征每个感应层，然后确认感应性能。标准PD-L1蛋白检测的校准图显示，两个动态范围为0.5-100.0 pm和100.0-500.0 pm，其中检测极限为0.20±0.001 pm（RSD≤5.2％），通过安培指定法为0.20±0.001 pm。通过检测A549肺癌细胞分泌的PD-L1和使用两种检测方法的可溶性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，通过对一小部分肺癌患者的SPD-L1水平进行定量研究了治疗试验。与健康个体（16.2-19.6 pm）相比，患者（221.6-240.4 pm）的SPD-L1水平明显更高。免疫疗法后，患者的PD-L1水平降至126.7-141.2 pm的范围。结果表明，使用两种建议的方法成功地进行了治疗监测。此外，基于对免疫检查点相关蛋白的比较研究，PD-L1比颗粒酶B和干扰素 - γ更有效。
Abstract
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.