Development of a SARS-CoV-2 nucleocapsid specific monoclonal antibody.

Development of a SARS-CoV-2 nucleocapsid specific monoclonal antibody.
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SARS-CoV-2核衣壳蛋白特异性单克隆抗体的研制

DOI:
10.1016/j.virol.2021.01.003
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发表时间:
2021-06
期刊:
影响因子:
3.7
通讯作者:
Geiss BJ
Geiss BJ
中科院分区:
医学3区
文献类型:
--
作者:
Terry JS;Anderson LB;Scherman MS;McAlister CE;Perera R;Schountz T;Geiss BJ

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为了帮助对抗 COVID-19,迫切需要专门针对 COVID-19 病原体 SARS-Coronavirus-2 (SARS-CoV-2) 关键成分的新分子工具。 SARS-CoV-2 核衣壳蛋白对于病毒复制至关重要,是病毒颗粒组装的组成部分,也是感染和免疫保护的主要诊断标记物。目前,针对核衣壳蛋白的有限可用抗体试剂并不特异于 SARS-CoV-2 核衣壳蛋白,并且这些抗体的序列尚未公开。在这项工作中,我们开发并表征了一系列针对 SARS-CoV-2 核衣壳蛋白的新型小鼠单克隆抗体,其中有一个特定克隆 mBG86,仅针对 SARS-CoV-2 核衣壳蛋白。单克隆抗体在 ELISA、蛋白质印迹和免疫荧光分析中得到验证。对来自六个选定克隆的可变区进行克隆和测序,并对测序克隆进行初步表位作图。总体而言,这些新型抗体试剂将在对抗 COVID-19 方面具有重要价值。
To help fight COVID-19, new molecular tools specifically targeting critical components of the causative agent of COVID-19, SARS-Coronavirus-2 (SARS-CoV-2), are desperately needed. The SARS-CoV-2 nucleocapsid protein is critical for viral replication, integral to viral particle assembly, and a major diagnostic marker for infection and immune protection. Currently the limited available antibody reagents targeting the nucleocapsid protein are not specific to SARS-CoV-2 nucleocapsid protein, and sequences for these antibodies are not publicly available. In this work we developed and characterized a series of new mouse monoclonal antibodies against the SARS-CoV-2 nucleocapsid protein, with a specific clone, mBG86, targeting only SARS-CoV-2 nucleocapsid protein. The monoclonal antibodies were validated in ELISA, Western blot, and immunofluorescence analyses. The variable regions from six select clones were cloned and sequenced, and preliminary epitope mapping of the sequenced clones was performed. Overall, these new antibody reagents will be of significant value in the fight against COVID-19.
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