Glycosylation-Targeting Aptamer for the Feasible Construction of a Dual Aptamer-Based Plasmonic Immunosandwich Assay in Cancer Diagnostics

IF 6.7 1区 化学 Q1 CHEMISTRY, ANALYTICAL
Junyang Chen, Pengfei Ma, Jiayu Xu, Mingxi Zang, Wei Li
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Abstract

Fibroblast activation protein (FAP) is an important antigen in the tumor microenvironment, which plays a crucial role in promoting extracellular matrix remodeling and tumor cell metastasis. A circulating form of soluble FAP has also been identified in the serum, becoming a biomarker for pan-cancer diagnosis and prognosis. However, the current peptide substrate-based enzymatic activity detection or antibody-dependent detection methods have been hindered by insufficient selectivity and complex operations, so it is valuable to develop effective nucleic acid aptamers as FAP affinity ligands. In order to deeply explore the biomimetic recognition technology, this study proposed an elaborate aptamer screening strategy for targeting the protein characteristic structure. Taking the glycosylation of the FAP protein as a target, four FAP-specific aptamers with high performance were successfully generated. Further, using the champion aptamer as a recognition tool and combining it with ultrasensitive detection technology-surface enhanced Raman scattering (SERS), a novel dual aptamer-based sandwich sensor was constructed for the rapid determination of FAP. Due to the dual-specific recognition of the orthogonal aptamer pair, the sandwich method obviously improved the selectivity to FAP protein, with a maximum cross-reactivity of less than 8% and a quantitation limit of 100 pg/mL. It was conveniently applied in high-sensitive and high-selective detection of serum FAP in cancer patient samples. Therefore, the research of this study not only opens new access for the selection of antiglycan aptamers but also boosts the application of the FAP aptamer as a recognition tool in cancer diagnostics.

Abstract Image

糖基化靶向适配体构建基于双适配体的肿瘤诊断等离子免疫夹心法
成纤维细胞活化蛋白(Fibroblast activation protein, FAP)是肿瘤微环境中的重要抗原,在促进细胞外基质重塑和肿瘤细胞转移中起着至关重要的作用。在血清中也发现了一种循环形式的可溶性FAP,成为泛癌诊断和预后的生物标志物。然而,目前基于肽底物的酶活性检测或依赖抗体的检测方法存在选择性不足、操作复杂等问题,因此开发有效的核酸适体作为FAP亲和配体具有重要意义。为了深入探索仿生识别技术,本研究针对蛋白质特征结构提出了一种精细的适配体筛选策略。以FAP蛋白的糖基化为靶点,成功生成了4个高性能的FAP特异性适配体。进一步,以该适配体为识别工具,结合超灵敏检测技术——表面增强拉曼散射(SERS),构建了一种新型的双适配体夹层传感器,用于FAP的快速测定。由于正交适体对的双特异性识别,夹层法明显提高了对FAP蛋白的选择性,最大交叉反应活性小于8%,定量限为100 pg/mL。该方法可方便地用于肿瘤患者血清FAP的高灵敏度、高选择性检测。因此,本研究的研究不仅为抗糖蛋白适配体的选择开辟了新的途径,而且促进了FAP适配体作为一种识别工具在癌症诊断中的应用。
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来源期刊
Analytical Chemistry
Analytical Chemistry 化学-分析化学
CiteScore
12.10
自引率
12.20%
发文量
1949
审稿时长
1.4 months
期刊介绍: Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.
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