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利用与CpG-DNA-脂质体复合物配制的合成肽表位生产靶向中东呼吸综合征冠状病毒M蛋白的单克隆抗体以检测中东呼吸综合征冠状病毒

Production of a Monoclonal Antibody Targeting the M Protein of MERS-CoV for Detection of MERS-CoV Using a Synthetic Peptide Epitope Formulated with a CpG-DNA-Liposome Complex.

作者信息

Park Byoung Kwon, Lee Su In, Bae Joon-Yong, Park Man-Seong, Lee Younghee, Kwon Hyung-Joo

机构信息

1Center for Medical Science Research, College of Medicine, Hallym University, Chuncheon, 24252 Republic of Korea.

2Department of Microbiology, College of Medicine, and the Institute for Viral Diseases, Korea University, Seoul, 02841 Republic of Korea.

出版信息

Int J Pept Res Ther. 2019;25(3):819-826. doi: 10.1007/s10989-018-9731-8. Epub 2018 Jul 2.

DOI:10.1007/s10989-018-9731-8
PMID:32214929
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7087630/
Abstract

The Middle East respiratory syndrome-related coronavirus (MERS-CoV) contains four major structural proteins, the spike glycoprotein, nucleocapsid phosphoprotein, membrane (M) glycoprotein and small envelope glycoprotein. The M protein of MERS-CoV has a role in the morphogenesis or assembly of the virus and inhibits type I interferon expression in infected cells. Here, we produced a monoclonal antibody specific against the M protein of MERS-CoV by injecting BALB/c mice with a complex containing the epitope peptide and CpG-DNA encapsulated with a phosphatidyl-β-oleoyl-γ-palmitoyl ethanolamine (DOPE):cholesterol hemisuccinate (CHEMS). The monoclonal antibody was reactive to the epitope peptide of the M protein of MERS-CoV which was confirmed by western blotting and immunoprecipitations. Indirect immunofluorescence assay and confocal image analysis showed that the monoclonal antibody binds specifically to the M protein of MERS-CoV in the virus-infected cells. Further studies using this monoclonal antibody may provide important information on the function of the M protein and its future application in diagnostics.

摘要

中东呼吸综合征相关冠状病毒（MERS-CoV）包含四种主要结构蛋白，即刺突糖蛋白、核衣壳磷蛋白、膜（M）糖蛋白和小囊膜糖蛋白。MERS-CoV的M蛋白在病毒的形态发生或组装中起作用，并抑制感染细胞中I型干扰素的表达。在此，我们通过向BALB/c小鼠注射一种复合物来制备针对MERS-CoV M蛋白的单克隆抗体，该复合物包含表位肽和用磷脂酰-β-油酰-γ-棕榈酰乙醇胺（DOPE）：胆固醇半琥珀酸酯（CHEMS）包裹的CpG-DNA。该单克隆抗体与MERS-CoV M蛋白的表位肽发生反应，这通过蛋白质免疫印迹法和免疫沉淀法得到证实。间接免疫荧光测定和共聚焦图像分析表明，该单克隆抗体在病毒感染细胞中特异性结合MERS-CoV的M蛋白。使用这种单克隆抗体的进一步研究可能会提供有关M蛋白功能及其在诊断中的未来应用的重要信息。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/9281fc2e138e/10989_2018_9731_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/396f8cbbd75c/10989_2018_9731_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/9fc9f1b656b3/10989_2018_9731_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/cee913aa791a/10989_2018_9731_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/9281fc2e138e/10989_2018_9731_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/396f8cbbd75c/10989_2018_9731_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/9fc9f1b656b3/10989_2018_9731_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/cee913aa791a/10989_2018_9731_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8ea2/7087630/9281fc2e138e/10989_2018_9731_Fig4_HTML.jpg

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