Terry James S, Anderson Loran Br, Scherman Michael S, McAlister Carley E, Perera Rushika, Schountz Tony, Geiss Brian J
Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA.
Arthropod-borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado, USA.
bioRxiv. 2020 Sep 3:2020.09.03.280370. doi: 10.1101/2020.09.03.280370.
The global COVID-19 pandemic has caused massive disruptions in every society around the world. 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 a major component of the viral replication processes, integral to viral particle assembly, and is a major diagnostic marker for infection and immune protection. Currently available antibody reagents targeting the nucleocapsid protein were primarily developed against the related SARS-CoV virus and are not specific to SARS-CoV-2 nucleocapsid protein. Therefore, in this work we developed and characterized a series of new mouse monoclonal antibodies against the SARS-CoV-2 nucleocapsid protein. The anti-nucleocapsid monoclonal antibodies were tested in ELISA, western blot, and immunofluorescence analyses. The variable regions from the heavy and light chains from five select clones were cloned and sequenced, and preliminary epitope mapping of the sequenced clones was performed. Overall, the new antibody reagents described here will be of significant value in the fight against COVID-19.
全球新冠疫情给世界各地的每个社会都带来了巨大破坏。为了帮助抗击新冠疫情，迫切需要专门针对新冠病毒病原体严重急性呼吸综合征冠状病毒2（SARS-CoV-2）关键成分的新型分子工具。SARS-CoV-2核衣壳蛋白是病毒复制过程的主要成分，对病毒颗粒组装不可或缺，并且是感染和免疫保护的主要诊断标志物。目前可用的针对核衣壳蛋白的抗体试剂主要是针对相关的SARS-CoV病毒开发的，并非特异性针对SARS-CoV-2核衣壳蛋白。因此，在这项工作中，我们开发并鉴定了一系列针对SARS-CoV-2核衣壳蛋白的新型小鼠单克隆抗体。这些抗核衣壳单克隆抗体在酶联免疫吸附测定（ELISA）、蛋白质免疫印迹法和免疫荧光分析中进行了测试。对五个选定克隆的重链和轻链可变区进行了克隆和测序，并对测序克隆进行了初步表位定位。总体而言，本文所述的新型抗体试剂在抗击新冠疫情中将具有重要价值。
