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基于免疫测定的生物传感器平台设计,用于使用高度特异性单克隆抗体检测新型冠状病毒。

Design of immunoassay based biosensor platforms for SARS-CoV-2 detection using highly specific monoclonal antibodies.

作者信息

Dinç Göknur Gizem, Saatçi Ebru, Polat İlkay Göksu, Yücel Fatıma, Tazebay Uygar Halis, Akçael Esin

机构信息

TÜBİTAK Marmara Research Center, 41470, Gebze, Kocaeli, Türkiye; Gebze Technical University, Department of Molecular Biology and Genetics, 41400, Gebze, Kocaeli, Türkiye.

Erciyes University, Faculty of Science, Department of Biology, 38039, Kayseri, Türkiye.

出版信息

Diagn Microbiol Infect Dis. 2025 Mar;111(3):116644. doi: 10.1016/j.diagmicrobio.2024.116644. Epub 2024 Dec 3.

Abstract

The global expand of SARS-CoV-2 has highlighted the importance of early and rapid detection to control the spread of a pandemic. In this study, specific and high-affinity monoclonal antibodies (mAbs) were developed against the conserved nucleocapsid protein of the virus among variants. Appropriate antibody pairs were selected to develop a lateral flow immunoassay (LFIA) and an unconventional application of an amperometric biosensor using unmodified screen-printed electrodes and external magnetic bead preparation. In the study, the LFIA we developed detected the SARS-CoV-2 virus at 10 PFU/mL, while the amperometric biosensor enabled sensitive detection of inactivated SARS-CoV-2 with an LOD of 5.5 PFU/mL. After validating the developed systems, it is considered that the mAbs we have obtained will enable the sensitive and selective detection of SARS-CoV-2 in LFIA and amperometric immunosensor platforms for clinical diagnosis.

摘要

严重急性呼吸综合征冠状病毒2(SARS-CoV-2)在全球的传播凸显了早期快速检测对于控制大流行传播的重要性。在本研究中,针对该病毒变异株中保守的核衣壳蛋白开发了特异性高亲和力单克隆抗体(mAb)。选择合适的抗体对开发了一种侧向流动免疫分析(LFIA)以及一种使用未修饰的丝网印刷电极和外部磁珠制备的电流型生物传感器的非常规应用。在该研究中,我们开发的LFIA能检测出10 PFU/mL的SARS-CoV-2病毒,而电流型生物传感器能够灵敏检测灭活的SARS-CoV-2,检测限为5.5 PFU/mL。在对开发的系统进行验证后,认为我们获得的单克隆抗体将能够在LFIA和电流型免疫传感器平台上灵敏且选择性地检测SARS-CoV-2用于临床诊断。

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