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

Preparation of highly specific monoclonal antibodies against SARS-CoV-2 nucleocapsid protein and the preliminary development of antigen detection test strips.
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DOI:
10.1002/jmv.27520
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发表时间:
2022-04
影响因子:
12.7
通讯作者:
Lv, Hangjun
Lv, Hangjun
中科院分区:
医学3区
文献类型:
--
作者:
Xie, Chengzuo;Ding, Haojie;Ding, Jianzu;Xue, Yangji;Lu, Shaohong;Lv, Hangjun

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2019年冠状病毒病(COVID - 19)正在全球暴发。为了帮助防治这种疾病,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)的核衣壳蛋白(N)参与病毒的复制、组装和免疫调节,在病毒生命周期中发挥重要作用。此外,N蛋白也可能是一个诊断因子和潜在的药物靶点。因此,我们通过合成SARS‐CoV‐2的N基因序列,构建pET‐28a(+)‐N重组质粒,在大肠杆菌中表达了N蛋白,并通过杂杂瘤和腹水获得了15个针对SARS‐CoV‐2‐N蛋白的单克隆抗体(mab),建立了检测N抗原的免疫层析试纸条方法。在这项研究中,我们获得了14种高效价和高特异性的单克隆抗体,并且试纸只与SARS - CoV - 2 - N蛋白发生反应,与其他冠状病毒无交叉反应,并且还识别Delta (B.1.617.2)变异的重组N蛋白。这些单克隆抗体可通过血清学抗原用于SARS - CoV - 2感染的早期快速诊断。
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.
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