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制备针对 SARS-CoV-2 核衣壳蛋白的高特异性单克隆抗体，并初步开发抗原检测试纸条。

Preparation of highly specific monoclonal antibodies against SARS-CoV-2 nucleocapsid protein and the preliminary development of antigen detection test strips.

机构信息

Institute of Parasitic Disease, Hangzhou Medical College, Hangzhou, China.

The Key Laboratory of Blood Safety Research, Blood Center of Zhejiang Province, Hangzhou, China.

出版信息

J Med Virol. 2022 Apr;94(4):1633-1640. doi: 10.1002/jmv.27520. Epub 2021 Dec 21.

DOI:10.1002/jmv.27520
PMID:34904253
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9303534/
Abstract

The coronavirus disease 2019 (COVID-19) is outbreaking all over the world. To help fight this disease, it is necessary to establish an effective and rapid detection method. The nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is involved in viral replication, assembly, and immune regulation and plays an important role in the viral life cycle. Moreover, the N protein also could be a diagnostic factor and potential drug target. Therefore, by synthesizing the N gene sequence of SARS-CoV-2, constructing the pET-28a (+)-N recombinant plasmid, we expressed the N protein in Escherichia coli and obtained 15 monoclonal antibody (mAbs) against SARS-CoV-2-N protein by the hybridomas and ascites, then an immunochromatographic test strip method detecting N antigen was established. In this study, we obtained 14 high-titer and high-specificity monoclonal antibodies, and the test strips exclusively react with the SARS-CoV-2-N protein and no cross-reactivity with other coronavirus and also recognize the recombinant N protein of Delta (B.1.617.2) variant. These mAbs can be used for the early and rapid diagnosis of SARS-CoV-2 infection through serological antigen.

摘要

新型冠状病毒肺炎（COVID-19）在全球范围内爆发。为了帮助对抗这种疾病，有必要建立一种有效和快速的检测方法。严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）的核衣壳（N）蛋白参与病毒复制、组装和免疫调节，在病毒生命周期中发挥重要作用。此外，N 蛋白也可以作为诊断因子和潜在的药物靶点。因此，通过合成 SARS-CoV-2 的 N 基因序列，构建 pET-28a（+）-N 重组质粒，我们在大肠杆菌中表达了 N 蛋白，并通过杂交瘤和腹水获得了 15 株针对 SARS-CoV-2-N 蛋白的单克隆抗体（mAbs），然后建立了一种检测 N 抗原的免疫层析测试条方法。在这项研究中，我们获得了 14 株高滴度和高特异性的单克隆抗体，该测试条仅与 SARS-CoV-2-N 蛋白反应，与其他冠状病毒无交叉反应，也可识别德尔塔（B.1.617.2）变异株的重组 N 蛋白。这些 mAbs 可通过血清学抗原用于 SARS-CoV-2 感染的早期和快速诊断。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32e2/9303534/b6a46459ccf7/JMV-94-1633-g007.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32e2/9303534/b6a46459ccf7/JMV-94-1633-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32e2/9303534/cb3776579e1b/JMV-94-1633-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32e2/9303534/a530f1d839d8/JMV-94-1633-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32e2/9303534/f8c628d9aee9/JMV-94-1633-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32e2/9303534/97b912cbfd5d/JMV-94-1633-g003.jpg
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