用于程序性死亡配体1（PD-L1）检测及癌症治疗进展监测的双信号输出生物传感器

Dual-signal output biosensor for the detection of program death-ligand 1 and therapy progress monitoring of cancer

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BIOSENSORS & BIOELECTRONICS

影响因子:10.5

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

DOI: 10.1016/j.bios.2024.116565

摘要

开发了一种一次性双输出生物传感器，用于检测程序性死亡配体1（PD-L1），以实现免疫治疗进展监测和早期癌症检测。该传感器采用与羧基化三苯乙烯聚合物（rGO-pTBA）复合的核糖核酸适配体（aptamer）探针，特异性捕获标记有新型氧化还原介体邻氨苯酚对硫酸的PD-L1蛋白，实现安培检测。每个传感层通过电化学和表面分析实验进行表征，确认其检测性能。标准PD-L1蛋白的校准曲线显示两个动态范围：0.5-100.0 pM和100.0-500.0 pM，检测极限为0.20±0.001 pM（RSD≤5.2%）。通过检测A549肺癌细胞分泌的PD-L1和临床相关的血清可溶性PD-L1（sPD-L1）水平验证了传感器的可靠性。还利用两种检测方法对少量肺癌患者进行了治疗效果的监测，发现患者（221.6-240.4 pM）血清中sPD-L1水平显著高于健康人（16.2-19.6 pM），免疫治疗后，该值下降至126.7-141.2 pM。结果表明，利用这两种方法成功实现了治疗监测。此外，通过对免疫检查点相关蛋白的比较研究发现，PD-L1作为生物标志物的效果优于颗粒酶B和干扰素γ。

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.