Xu Xin, Zhang Ze, Yu Hongwei, Shen Tong, Pan Hongzhi, Chang Dong
Department of Clinical Laboratory, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai 201399, China.
Hebei Medical University, Shijiazhuang,Hebei 050011, China.
ACS Omega. 2024 Aug 2;9(32):34493-34506. doi: 10.1021/acsomega.4c02346. eCollection 2024 Aug 13.
Nonsmall cell lung cancer (NSCLC), due to its lack of early symptoms, has become one of the leading causes of cancer-related deaths globally. Exosomes, small membrane vesicles secreted by cells, are widely present in human bodily fluids. In the bodily fluids of NSCLC patients, the quantity of extracellular vesicles is double that of healthy individuals, suggesting their potential as biomarkers for screening NSCLC. This study designed a dual-modal aptasensor that integrated excellent sensitivity in electrochemical detection and portability in fluorescence detection into one device. AuNPs were functionalized with exosome-capturing probes containing thiol-modified CD63 aptamers, which were immobilized on screen-printed gold electrodes. On the other hand, the carboxylated CD63 aptamer was immobilized on the surface of PB-modified g-CN loaded with Co-SANs particles (Co@g-CN@PB). By combining these components, a sandwich structure (AuNPs/Apt1/Exo/Apt2- Co@g-CN@PB) was constructed, forming a probe for specific exosome recognition. First, the samples were preliminarily assessed for their positive or negative status under a fluorescence inverted microscope. Subsequently, a more in-depth quantitative analysis was conducted on suspected positive samples using electrochemical or fluorescence analysis methods. The detection limits for electrochemical analysis and fluorescence analysis were 66.68 and 33.5particles/mL, respectively. In the analysis of clinical serum exosome samples, the developed dual-modal aptasensor effectively distinguished serum specimens from those of NSCLC patients and healthy volunteers. This highlighted the inspection capability of the dual-modal adapter sensor, especially in point-of-care testing, making it a highly suitable tool for clinical applications.
非小细胞肺癌（NSCLC）由于缺乏早期症状，已成为全球癌症相关死亡的主要原因之一。外泌体是细胞分泌的小膜泡，广泛存在于人体体液中。在NSCLC患者的体液中，细胞外囊泡的数量是健康个体的两倍，这表明它们作为NSCLC筛查生物标志物的潜力。本研究设计了一种双模态适体传感器，将电化学检测中的高灵敏度和荧光检测中的便携性集成到一个设备中。金纳米颗粒（AuNPs）用含有硫醇修饰的CD63适体的外泌体捕获探针进行功能化，该探针固定在丝网印刷金电极上。另一方面，羧基化的CD63适体固定在负载有Co-SANs颗粒的PB修饰的g-CN表面（Co@g-CN@PB）。通过组合这些组件，构建了一种三明治结构（AuNPs/Apt1/Exo/Apt2-Co@g-CN@PB），形成了一种用于特异性外泌体识别的探针。首先，在荧光倒置显微镜下对样品的阳性或阴性状态进行初步评估。随后，使用电化学或荧光分析方法对疑似阳性样品进行更深入的定量分析。电化学分析和荧光分析的检测限分别为66.68和33.5颗粒/毫升。在临床血清外泌体样品分析中，所开发的双模态适体传感器有效地区分了NSCLC患者和健康志愿者的血清标本。这突出了双模态适配传感器的检测能力，特别是在即时检测中，使其成为临床应用的高度合适工具。
