Center for Proteomics, Faculty of Medicine, University of Rijeka, Braće Branchetta 20, 51000 Rijeka, Croatia.
Research Department, University Hospital for Infectious Diseases "Dr. Fran Mihaljević", Mirogojska 8, 10000 Zagreb, Croatia.
Viruses. 2022 Feb 21;14(2):443. doi: 10.3390/v14020443.
In early 2020, the COVID-19 pandemic sparked a global crisis that continues to pose a serious threat to human health and the economy. Further advancement in research is necessary and requires the availability of quality molecular tools, including monoclonal antibodies. Here, we present the development and characterization of a collection of over 40 new monoclonal antibodies directed against different SARS-CoV-2 proteins. Recombinant SARS-CoV-2 proteins were expressed, purified, and used as immunogens. Upon development of specific hybridomas, the obtained monoclonal antibody (mAb) clones were tested for binding to recombinant proteins and infected cells. We generated mAbs against structural proteins, the Spike and Nucleocapsid protein, several non-structural proteins (nsp1, nsp7, nsp8, nsp9, nsp10, nsp16) and accessory factors (ORF3a, ORF9b) applicable in flow cytometry, immunofluorescence, or Western blot. Our collection of mAbs provides a set of novel, highly specific tools that will allow a comprehensive analysis of the viral proteome, which will allow further understanding of SARS-CoV-2 pathogenesis and the design of therapeutic strategies.
2020 年初，COVID-19 大流行引发了一场全球危机，继续对人类健康和经济构成严重威胁。进一步的研究进展是必要的，需要有高质量的分子工具，包括单克隆抗体。在这里，我们介绍了一组针对不同 SARS-CoV-2 蛋白的 40 多种新的单克隆抗体的开发和特性。重组 SARS-CoV-2 蛋白被表达、纯化，并用作免疫原。在开发出特定的杂交瘤后，对获得的单克隆抗体 (mAb) 克隆进行了测试，以检测其与重组蛋白和感染细胞的结合情况。我们生成了针对结构蛋白（Spike 和 Nucleocapsid 蛋白）、几种非结构蛋白（nsp1、nsp7、nsp8、nsp9、nsp10、nsp16）和辅助因子（ORF3a、ORF9b）的 mAbs，可用于流式细胞术、免疫荧光或 Western blot。我们的 mAb 集合提供了一套新的、高度特异性的工具，可用于全面分析病毒蛋白质组，从而进一步了解 SARS-CoV-2 的发病机制和治疗策略的设计。
