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建立产生针对 SARS-CoV-2 刺突蛋白抗体的小鼠杂交瘤细胞。

Establishment of Murine Hybridoma Cells Producing Antibodies against Spike Protein of SARS-CoV-2.

机构信息

Department of Functioning of Living Systems, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, 117997 Moscow, Russia.

Institute of Medicine, Peoples' Friendship University of Russia (RUDN University), 17198 Moscow, Russia.

出版信息

Int J Mol Sci. 2020 Dec 1;21(23):9167. doi: 10.3390/ijms21239167.

Abstract

In 2020 the world faced the pandemic of COVID-19 severe acute respiratory syndrome caused by a new type of coronavirus named SARS-CoV-2. To stop the spread of the disease, it is crucial to create molecular tools allowing the investigation, diagnoses and treatment of COVID-19. One of such tools are monoclonal antibodies (mAbs). In this study we describe the development of hybridoma cells that can produce mouse mAbs against receptor binding domain of SARS-CoV-2 spike (S) protein. These mAbs are able to specifically detect native and denatured S proteins in all tested applications, including immunoblotting, enzyme-linked immunosorbent assay, immunofluorescence staining of cells and immunohistochemical staining of paraffin embedded patients' tissue samples. In addition, we showed that the obtained mAbs can efficiently block SARS-CoV-2 infection in in vitro experiments. Finally, we determined the amino acid sequence of light and heavy chains of the mAbs. This information will allow the use of corresponding peptides to establish genetically engineered therapeutic antibodies. To date multiple mAbs against SARS-CoV-2 proteins have been established, however, bigger sets of various antibodies will allow the detection and neutralization of SARS-CoV-2, even if the virus acquires novel mutations.

摘要

2020 年,世界面临由新型冠状病毒 SARS-CoV-2 引起的 COVID-19 严重急性呼吸综合征大流行。为了阻止疾病的传播,开发能够用于 COVID-19 调查、诊断和治疗的分子工具至关重要。其中一种工具是单克隆抗体 (mAbs)。在这项研究中,我们描述了杂交瘤细胞的开发,这些细胞可以产生针对 SARS-CoV-2 刺突 (S) 蛋白受体结合域的鼠 mAbs。这些 mAbs 能够在所有测试应用中特异性检测天然和变性的 S 蛋白,包括免疫印迹、酶联免疫吸附试验、细胞免疫荧光染色和石蜡包埋患者组织样本的免疫组织化学染色。此外,我们表明,获得的 mAbs 可在体外实验中有效阻止 SARS-CoV-2 感染。最后,我们确定了 mAbs 的轻链和重链的氨基酸序列。该信息将允许使用相应的肽来建立基因工程治疗性抗体。迄今为止,已经建立了多种针对 SARS-CoV-2 蛋白的 mAbs,但是更大的各种抗体集将允许检测和中和 SARS-CoV-2,即使病毒发生新的突变。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d2e7/7731011/da33072e2fa7/ijms-21-09167-g001.jpg

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