基于MoS2@AuNPs组装双肽作为生物探针，具有显著的双信号放大的病原体抗原敏感安培免疫传感器

IF 5.7 2区化学 Q1 CHEMISTRY, ANALYTICAL

Analytica Chimica Acta Pub Date : 2025-04-04 DOI:10.1016/j.aca.2025.344015

Zongmei Zheng , Mingyang Wang , Jinlong Yuan , Junchong Liu , Haipeng Yu , Zhonghuang Yang , Wanjian Liu , Aihua Liu

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引用次数: 0

摘要

及时准确识别致病病毒，对早期治疗和紧急预防至关重要。病毒抗原检测可以识别那些最有可能传播疾病的人，但大多数是基于稳定性和敏感性有限的抗体。与抗体相比，多肽具有成本低、体积小、稳定性好等优点。利用特异性肽探针进行电化学免疫分析具有灵敏度、选择性好、稳定性好等优点。本文报道了一种利用多肽作为捕获探针和识别探针的安培免疫传感器。分子对接表明，从噬菌体展示中生物筛选的Pi（序列：nfwisplklafalgggkkkksc）和FK11（序列：WFLNDSELISML）的两个肽结合到SARS-CoV-2刺突蛋白（SP）的n端结构域。将肽Pi组装在MoS2@AuNPs修饰电极上捕获SARS-CoV-2 SP，被肽fk11展示的噬菌体识别，形成Pi/SARS-CoV-2 SP/ fk11噬菌体三明治。然后引入抗M13噬菌体偶联的辣根过氧化物酶（HRP）（anti M13-HRP）识别噬菌体衣壳蛋白pVIII，形成M13噬菌体/抗M13-HRP，富集数千个HRP，进一步在- 0.35 V高导电性MoS2@AuNPs下电化学催化H2O2还原。构建免疫传感器，线性范围为0.1 ~ 5000 pg/mL，检出限为0.074 pg/mL。该传感器还具有良好的选择性、批量重现性和稳定性，可检测到10个转导单位/mL的SARS-CoV-2假病毒。本研究首次实现了基于双肽探针的三明治型电化学免疫传感器与双信号放大的集成，为检测真正的SARS-CoV-2病毒提供了一个具有成本效益的检测平台，可用于早期诊断。灵活和模块化的策略可以扩展到开发其他类型的生物传感器的广泛应用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Sensitive amperometric immunosensor for pathogen antigen based on MoS2@AuNPs assembling dual-peptide as bioprobes with significant dual signal amplification

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Sensitive amperometric immunosensor for pathogen antigen based on MoS2@AuNPs assembling dual-peptide as bioprobes with significant dual signal amplification

It is crucial to timely and accurately identify the causative virus for early treatment and urgent prevention. Viral antigen detection can identify those people who are most likely at risk of spreading the disease, but most based on antibodies with limited stability and sensitivity. Peptides offer several advantages over antibodies, such as low cost, smaller size and good stability. The development of electrochemical immunoassay using specific peptide probes have the merits of good sensitivity and selectivity as well as good stability. Herein we report an amperometric immunosensor using peptides as capture probe and recognition probe. The molecular docking suggests that the two peptides of Pi (sequence: NFWISPKLAFALGGGKKKSC) and FK11 (sequence: WFLNDSELISML), bioscreened from phage display, bind to N-terminal domain of SARS-CoV-2 spike protein (SP). The peptide Pi is assembled on MoS₂@AuNPs modified electrode to capture SARS-CoV-2 SP, which is recognized by peptide FK11-displayed phage to form Pi/SARS-CoV-2 SP/FK11-phage sandwich. Then anti-M13 phage conjugated horseradish peroxidase (HRP) (anti M13-HRP) was introduced to recognize the phage capsid protein pVIII to form M13 phage/anti M13-HRP to enrich thousands of HRP, which can further electrochemically catalyze H₂O₂ reduction at highly conductive MoS₂@AuNPs at − 0.35 V. Then amperometric immunosensor was constructed with linear range of 0.1–5000 pg/mL SARS-CoV-2 SP and detection limit of 0.074 pg/mL. The sensor also has good selectivity, batch reproducibility and stability, capable of detecting down to 10 transducing units/mL SARS-CoV-2 pseudoviruses. This work represents the first example of dual-peptide probes based sandwich-type electrochemical immunosensor integrated with dual signal amplification, which may provide a cost-effective assay platform in detecting real SARS-CoV-2 viruses for early diagnosis. The flexible and modular strategy can be extended to develop other type biosensors for a wide range of applications.

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来源期刊

Analytica Chimica Acta 化学-分析化学

CiteScore

10.40

自引率

6.50%

发文量

1081

审稿时长

38 days

期刊介绍： Analytica Chimica Acta has an open access mirror journal Analytica Chimica Acta: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. Analytica Chimica Acta provides a forum for the rapid publication of original research, and critical, comprehensive reviews dealing with all aspects of fundamental and applied modern analytical chemistry. The journal welcomes the submission of research papers which report studies concerning the development of new and significant analytical methodologies. In determining the suitability of submitted articles for publication, particular scrutiny will be placed on the degree of novelty and impact of the research and the extent to which it adds to the existing body of knowledge in analytical chemistry.