使用截短的适配体和 SYBR 绿 I 无标记检测程序性死亡配体 1 以进行预后分析

IF 2.6 4区 化学 Q2 BIOCHEMICAL RESEARCH METHODS Journal of Fluorescence Pub Date : 2024-10-01 DOI:10.1007/s10895-024-03960-x
Ling Li, Zheng-Ling Li, Wei Wang, Xin-Xin Long, Ping Liu, Guo-Tian Wang, Shi-Qing Sun, Rong-Lian Du
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引用次数: 0

摘要

快速准确地检测程序性死亡配体1(PD-L1)的表达对肿瘤的诊断和治疗具有重要价值。基于ELISA的传统方法是蛋白质检测的金标准,但仍存在一些缺陷,尤其是抗原抗体依赖性,大大增加了检测时间和成本。本研究以截短的aptamer为识别分子,以双链DNA特异性染料(SYBR Green I)为信号单元,构建了一种快速灵敏检测PD-L1的无标记荧光探针。经过一系列条件优化,该探针对缓冲溶液中的 PD-L1 具有良好的检测能力,检测限低至 0.68 ng/mL。由于适配体与目标物的特异性识别能力,该方法对 PD-L1 的检测也具有良好的选择性。人血清样本中 PD-L1 的回收率为 86.20% 至 96.36%。与其他方法相比,该策略无需标记,不需要其他复杂的设计和纯化过程,操作过程简单,抗干扰能力强。整个检测过程可在 20 分钟内完成,具有良好的应用前景。这项工作将为特异性肿瘤治疗的药物剂量和预后评估提供参考。
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Label-Free Detection of Programmed Death-Ligand 1 for Prognosis Using a Truncated Aptamer and SYBR Green I.

The rapid and accurate detection of programmed death-ligand 1 (PD-L1) expression is of great value in the diagnosis and treatment of tumors. ELISA-based traditional method is the gold standard for protein detection, but there are still some shortcomings, especially the antigen-antibody dependence, greatly increased the detection time and cost. This work constructed a label-free fluorescent probe for rapid and sensitive detection of PD-L1 using a truncated aptamer as recognition molecules and double-stranded DNA specific dyes (SYBR Green I) as signal units. After a series of optimization conditions, this probe has good detection capability for PD-L1 in buffer solution with the detection limit as low as 0.68 ng/mL. Due to the specific recognition ability of aptamer and target, this method also has good selectivity for PD-L1 detection. The recovery of PD-L1 in human serum samples ranges from 86.20 to 96.36%. Compared with other methods, this strategy does not need to be marked, and does not need other complex design and purification process, but simple operation process and strong anti-interference ability. The whole detection process can be completed within 20 min and has good application prospect. This work will provide reference for drug dosage and prognosis evaluation of specific tumor therapy.

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来源期刊
Journal of Fluorescence
Journal of Fluorescence 化学-分析化学
CiteScore
4.60
自引率
7.40%
发文量
203
审稿时长
5.4 months
期刊介绍: Journal of Fluorescence is an international forum for the publication of peer-reviewed original articles that advance the practice of this established spectroscopic technique. Topics covered include advances in theory/and or data analysis, studies of the photophysics of aromatic molecules, solvent, and environmental effects, development of stationary or time-resolved measurements, advances in fluorescence microscopy, imaging, photobleaching/recovery measurements, and/or phosphorescence for studies of cell biology, chemical biology and the advanced uses of fluorescence in flow cytometry/analysis, immunology, high throughput screening/drug discovery, DNA sequencing/arrays, genomics and proteomics. Typical applications might include studies of macromolecular dynamics and conformation, intracellular chemistry, and gene expression. The journal also publishes papers that describe the synthesis and characterization of new fluorophores, particularly those displaying unique sensitivities and/or optical properties. In addition to original articles, the Journal also publishes reviews, rapid communications, short communications, letters to the editor, topical news articles, and technical and design notes.
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