Purification and characterization of the receptor-binding domain of SARS-CoV-2 spike protein from Escherichia coli

doi:10.1002/elsc.202000106

	Purification and characterization of the receptor-binding domain of SARS-CoV-2 spike protein from Escherichia coli
	He, Yunxia 1,2; Qi, Jinming 1,2; Xiao, Lucheng 1,2; Shen, Lijuan 2; Yu, Weili 2; Hu, Tao 2
刊名	ENGINEERING IN LIFE SCIENCES
	2021-05-07
页码	8
关键词	COVID‐ 19 purification RBD SARS‐ CoV‐ 2 spike protein
ISSN号	1618-0240
DOI	10.1002/elsc.202000106
英文摘要	SARS-CoV-2 is responsible for a disruptive worldwide viral pandemic, and renders a severe respiratory disease known as COVID-19. Spike protein of SARS-CoV-2 mediates viral entry into host cells by binding ACE2 through the receptor-binding domain (RBD). RBD is an important target for development of virus inhibitors, neutralizing antibodies, and vaccines. RBD expressed in mammalian cells suffers from low expression yield and high cost. E. coli is a popular host for protein expression, which has the advantage of easy scalability with low cost. However, RBD expressed by E. coli (RBD-1) lacks the glycosylation, and its antigenic epitopes may not be sufficiently exposed. In the present study, RBD-1 was expressed by E. coli and purified by a Ni Sepharose Fast Flow column. RBD-1 was structurally characterized and compared with RBD expressed by the HEK293 cells (RBD-2). The secondary structure and tertiary structure of RBD-1 were largely maintained without glycosylation. In particular, the major beta-sheet content of RBD-1 was almost unaltered. RBD-1 could strongly bind ACE2 with a dissociation constant (K-D) of 2.98 x 10(-8) M. Thus, RBD-1 was expected to apply in the vaccine development, screening drugs and virus test kit.
资助项目	National Key Research and Development Project ofChina ; NationalNatural Science Foundation of China[31970875] ; BeijingNatural Science Foundation
WOS关键词	EXPRESSION
WOS研究方向	Biotechnology & Applied Microbiology
语种	英语
出版者	WILEY
WOS记录号	WOS:000648189700001
资助机构	National Key Research and Development Project ofChina ; NationalNatural Science Foundation of China ; BeijingNatural Science Foundation
内容类型	期刊论文
源URL	[http://ir.ipe.ac.cn/handle/122111/48637]
专题	中国科学院过程工程研究所
通讯作者	Yu, Weili; Hu, Tao
作者单位	1.Univ Chinese Acad Sci, Beijing, Peoples R China 2.Chinese Acad Sci, Inst Proc Engn, State Key Lab Biochem Engn, Beijing 100190, Peoples R China
推荐引用方式 GB/T 7714	He, Yunxia,Qi, Jinming,Xiao, Lucheng,et al. Purification and characterization of the receptor-binding domain of SARS-CoV-2 spike protein from Escherichia coli[J]. ENGINEERING IN LIFE SCIENCES,2021:8.
APA	He, Yunxia,Qi, Jinming,Xiao, Lucheng,Shen, Lijuan,Yu, Weili,&Hu, Tao.(2021).Purification and characterization of the receptor-binding domain of SARS-CoV-2 spike protein from Escherichia coli.ENGINEERING IN LIFE SCIENCES,8.
MLA	He, Yunxia,et al."Purification and characterization of the receptor-binding domain of SARS-CoV-2 spike protein from Escherichia coli".ENGINEERING IN LIFE SCIENCES (2021):8.