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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Production, Characterization, and Uses of Monoclonal Antibodies against Recombinant Nucleoprotein of Elk Coronavirus
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Production, Characterization, and Uses of Monoclonal Antibodies against Recombinant Nucleoprotein of Elk Coronavirus

机译：麋鹿冠状病毒重组核蛋白单克隆抗体的生产，鉴定和应用

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This is the first report of the production of monoclonal antibodies against elk coronavirus. The nucleoprotein gene of elk coronavirus was amplified by PCR and was cloned and expressed in a prokaryotic expression vector. Recombinant nucleocapsid protein was used to immunize mice for the production of hybridomas. Twelve hybridomas that produced monoclonal antibodies against the nucleocapsid protein of elk coronavirus were selected by an indirect fluorescent-antibody test, an enzyme-linked immunosorbent assay, and a Western blot assay. Ten of the monoclonal antibodies were of the immunoglobulin G1 (IgG1) isotype, one was IgG2a, and one was IgM. All had kappa light chains. By immunohistochemistry four monoclonal antibodies detected bovine coronavirus and elk coronavirus in formalin-fixed intestinal tissues. Antinucleoprotein monoclonal antibodies were found to be better at ruminant coronavirus detection than the anti-spike protein monoclonal antibodies. Because nucleoprotein is a more abundant antigen than spike protein in infected cells, this was not an unexpected finding.
机译：这是针对麋鹿冠状病毒的单克隆抗体生产的首次报道。通过PCR扩增麋鹿冠状病毒的核蛋白基因，并将其克隆并在原核表达载体中表达。重组核衣壳蛋白用于免疫小鼠以产生杂交瘤。通过间接荧光抗体试验，酶联免疫吸附试验和蛋白质印迹试验选择了十二种产生针对麋鹿冠状病毒核衣壳蛋白的单克隆抗体的杂交瘤。十种单克隆抗体具有免疫球蛋白G1（IgG1）同种型，一种为IgG2a，一种为IgM。所有人都有κ轻链。通过免疫组织化学，四种单克隆抗体在福尔马林固定的肠组织中检测到牛冠状病毒和麋鹿冠状病毒。在反刍冠状病毒检测中发现抗核蛋白单克隆抗体比抗穗蛋白单克隆抗体更好。因为在感染的细胞中核蛋白是一种比刺突蛋白更丰富的抗原，所以这并不是一个意外的发现。

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