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在本氏烟中快速生产 SARS-CoV-2 受体结合域(RBD)和刺突蛋白特异性单克隆抗体 CR3022。

Rapid production of SARS-CoV-2 receptor binding domain (RBD) and spike specific monoclonal antibody CR3022 in Nicotiana benthamiana.

机构信息

Research Unit for Plant-Produced Pharmaceuticals, Chulalongkorn University, Bangkok, Thailand.

Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.

出版信息

Sci Rep. 2020 Oct 19;10(1):17698. doi: 10.1038/s41598-020-74904-1.

Abstract

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is responsible for the ongoing global outbreak of coronavirus disease (COVID-19) which is a significant threat to global public health. The rapid spread of COVID-19 necessitates the development of cost-effective technology platforms for the production of vaccines, drugs, and protein reagents for appropriate disease diagnosis and treatment. In this study, we explored the possibility of producing the receptor binding domain (RBD) of SARS-CoV-2 and an anti-SARS-CoV monoclonal antibody (mAb) CR3022 in Nicotiana benthamiana. Both RBD and mAb CR3022 were transiently produced with the highest expression level of 8 μg/g and 130 μg/g leaf fresh weight respectively at 3 days post-infiltration. The plant-produced RBD exhibited specific binding to the SARS-CoV-2 receptor, angiotensin-converting enzyme 2 (ACE2). Furthermore, the plant-produced mAb CR3022 binds to SARS-CoV-2, but fails to neutralize the virus in vitro. This is the first report showing the production of anti-SARS-CoV-2 RBD and mAb CR3022 in plants. Overall these findings provide a proof-of-concept for using plants as an expression system for the production of SARS-CoV-2 antigens and antibodies or similar other diagnostic reagents against SARS-CoV-2 rapidly, especially during epidemic or pandemic situation.

摘要

严重急性呼吸系统综合症冠状病毒 2 型(SARS-CoV-2)是引发目前全球冠状病毒疾病(COVID-19)大流行的主要原因,对全球公共健康构成重大威胁。COVID-19 的快速传播需要开发具有成本效益的技术平台,用于生产疫苗、药物和蛋白质试剂,以进行适当的疾病诊断和治疗。在本研究中,我们探索了在本氏烟中生产 SARS-CoV-2 的受体结合域(RBD)和抗 SARS-CoV 单克隆抗体(mAb)CR3022 的可能性。RBD 和 mAb CR3022 均在瞬时表达,在侵染后第 3 天达到最高表达水平,分别为 8μg/g 和 130μg/g 叶片鲜重。植物表达的 RBD 特异性结合 SARS-CoV-2 的受体血管紧张素转化酶 2(ACE2)。此外,植物产生的 mAb CR3022 与 SARS-CoV-2 结合,但不能在体外中和该病毒。这是首次报道在植物中生产抗 SARS-CoV-2 的 RBD 和 mAb CR3022。总的来说,这些发现为使用植物作为 SARS-CoV-2 抗原和抗体或针对 SARS-CoV-2 的类似其他诊断试剂的表达系统提供了概念验证,特别是在疫情或大流行期间,可以快速生产。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ca43/7573609/dd8de9ca6dc7/41598_2020_74904_Fig1_HTML.jpg

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