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特异性识别新型冠状病毒刺突蛋白受体结合域线性表位的兔单克隆抗体。

Rabbit Monoclonal Antibody Specifically Recognizing a Linear Epitope in the RBD of SARS-CoV-2 Spike Protein.

作者信息

Hong Junping, Wang Qian, Wu Qian, Chen Junyu, Wang Xijing, Wang Yingbin, Chen Yixin, Xia Ningshao

机构信息

National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen 361102, China.

State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, School of Public Health, Xiamen University, Xiamen 361102, China.

出版信息

Vaccines (Basel). 2021 Jul 28;9(8):829. doi: 10.3390/vaccines9080829.

Abstract

To date, SARS-CoV-2 pandemic has caused more than 188 million infections and 4.06 million deaths worldwide. The receptor-binding domain (RBD) of the SARS-CoV-2 spike protein has been regarded as an important target for vaccine and therapeutics development because it plays a key role in binding the human cell receptor ACE2 that is required for viral entry. However, it is not easy to detect RBD in Western blot using polyclonal antibody, suggesting that RBD may form a complicated conformation under native condition and bear rare linear epitope. So far, no linear epitope on RBD is reported. Thus, a monoclonal antibody (mAb) that recognizes linear epitope on RBD will become valuable. In the present study, an RBD-specific rabbit antibody named 9E1 was isolated from peripheral blood mononuclear cells (PBMC) of immunized rabbit by RBD-specific single B cell sorting and mapped to a highly conserved linear epitope within twelve amino acids CNGVEGFNCYFP on RBD. 9E1 works well in Western blot on S protein and immunohistochemistry on the SARS-CoV-2 infected tissue sections. The results demonstrated that 9E1 can be used as a useful tool for pathological and functional studies of SARS-CoV-2.

摘要

迄今为止,新型冠状病毒肺炎大流行已在全球造成超过1.88亿人感染和406万人死亡。新型冠状病毒刺突蛋白的受体结合域(RBD)被视为疫苗和治疗药物开发的重要靶点,因为它在结合病毒进入所需的人类细胞受体血管紧张素转换酶2(ACE2)中起关键作用。然而,使用多克隆抗体在蛋白质免疫印迹法中检测RBD并不容易,这表明RBD在天然条件下可能形成复杂的构象,且带有罕见的线性表位。到目前为止,尚未报道RBD上的线性表位。因此,一种识别RBD上线性表位的单克隆抗体(mAb)将变得很有价值。在本研究中,通过RBD特异性单B细胞分选从免疫兔的外周血单个核细胞(PBMC)中分离出一种名为9E1的RBD特异性兔抗体,并将其定位到RBD上十二个氨基酸CNGVEGFNCYFP内的一个高度保守的线性表位。9E1在S蛋白的蛋白质免疫印迹法和新型冠状病毒感染组织切片的免疫组织化学中表现良好。结果表明,9E1可作为新型冠状病毒病理和功能研究的有用工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2fb/8402368/7a3a3c2a9b8d/vaccines-09-00829-g001.jpg

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