Efficient production of recombinant SARS-CoV-2 spike protein using the baculovirus-silkworm system

被引:39
作者
Fujita, Ryosuke [1 ]
Hino, Masato [1 ]
Ebihara, Takeru [2 ]
Nagasato, Takumi [2 ]
Masuda, Akitsu [2 ]
Lee, Jae Man [3 ]
Fujii, Tsuguru [3 ]
Mon, Hiroaki [4 ]
Kakino, Kohei [2 ]
Nagai, Ryo [2 ]
Tanaka, Miyu [2 ]
Tonooka, Yoshino [2 ]
Moriyama, Takato [2 ]
Kusakabe, Takahiro [4 ]
机构
[1] Kyushu Univ, Fac Agr, Lab Sanit Entomol, Nishi Ku, Motooka 744, Fukuoka 8190395, Japan
[2] Kyushu Univ, Grad Sch Bioresource & Bioenvironm Sci, Lab Insect Genome Sci, Nishi Ku, Motooka 744, Fukuoka 8190395, Japan
[3] Kyushu Univ, Fac Agr, Lab Creat Sci Insect Ind, Nishi Ku, Motooka 744, Fukuoka 8190395, Japan
[4] Kyushu Univ, Fac Agr, Lab Insect Genome Sci, Nishi Ku, Motooka 744, Fukuoka 8190395, Japan
关键词
Silkworm-BmNPV expression system; SARS-CoV-2; Spike protein; EXPRESSION; VACCINE; VIRUS; ENTRY;
D O I
10.1016/j.bbrc.2020.06.020
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
In the case of a new viral disease outbreak, an immediate development of virus detection kits and vaccines is required. For COVID-19, we established a rapid production procedure for SARS-CoV-2 spike protein (S protein) by using the baculovirus-silkworm expression system. The baculovirus vector-derived S proteins were successfully secreted to silkworm serum, whereas those formed insoluble structure in the larval fat body and the pupal cells. The ectodomain of S protein with the native sequence was cleaved by the host furin-protease, resulting in less recombinant protein production. The S protein modified in furin protease-target site was efficiently secreted to silkworm serum and was purified as oligomers, which showed immunoreactivity for anti-SARS-CoV-2 S2 antibody. By using the direct transfection of recombinant bacmid to silkworms, we achieved the efficient production of SARS-CoV-2 S protein as fetal bovine serum (FBS)-free system. The resultant purified S protein would be useful tools for the development of immunodetection kits, antigen for immunization for immunoglobulin production, and vaccines. (C) 2020 Published by Elsevier Inc.
引用
收藏
页码:257 / 262
页数:6
相关论文
共 21 条
[1]   Development of epitope-based peptide vaccine against novel coronavirus 2019 (SARS-COV-2): Immunoinformatics approach [J].
Bhattacharya, Manojit ;
Sharma, Ashish R. ;
Patra, Prasanta ;
Ghosh, Pratik ;
Sharma, Garima ;
Patra, Bidhan C. ;
Lee, Sang-Soo ;
Chakraborty, Chiranjib .
JOURNAL OF MEDICAL VIROLOGY, 2020, 92 (06) :618-631
[2]   The spike glycoprotein of the new coronavirus 2019-nCoV contains a furinlike cleavage site absent in CoV of the same clade [J].
Coutard, B. ;
Valle, C. ;
de lamballerie, X. ;
Canard, B. ;
Seidah, N. G. ;
Decroly, E. .
ANTIVIRAL RESEARCH, 2020, 176
[3]   Development of a rapid test kit for SARS-CoV-2: an example of product design [J].
Cui, Zhanfeng ;
Chang, Hong ;
Wang, Hui ;
Lim, Boon ;
Hsu, Chia-Chen ;
Yu, Yejiong ;
Jia, Huidong ;
Wang, Yun ;
Zeng, Yida ;
Ji, Mengmeng ;
Liu, Weizhi ;
Inverarity, Catriona ;
Huang, Wei E. .
BIO-DESIGN AND MANUFACTURING, 2020, 3 (02) :83-86
[4]   The Challenges of Vaccine Development against a New Virus during a Pandemic [J].
Diamond, Michael S. ;
Pierson, Theodore C. .
CELL HOST & MICROBE, 2020, 27 (05) :699-703
[5]   Fiber knob domain lacking the shaft sequence but fused to a coiled coil is a candidate subunit vaccine against egg-drop syndrome [J].
Harakuni, Tetsuya ;
Andoh, Kiyohiko ;
Sakamoto, Ryu-ichi ;
Tamaki, Yukihiro ;
Miyata, Takeshi ;
Uefuji, Hirotaka ;
Yamazaki, Ken-ichi ;
Arakawa, Takeshi .
VACCINE, 2016, 34 (27) :3184-3190
[6]   Phylogenetic Analysis and Structural Modeling of SARS-CoV-2 Spike Protein Reveals an Evolutionary Distinct and Proteolytically Sensitive Activation Loop [J].
Jaimes, Javier A. ;
Andre, Nicole M. ;
Chappie, Joshua S. ;
Millet, Jean K. ;
Whittaker, Gary R. .
JOURNAL OF MOLECULAR BIOLOGY, 2020, 432 (10) :3309-3325
[7]   Expression and purification of biologically active human granulocyte-macrophage colony stimulating factor (hGM-CSF) using silkworm-baculovirus expression vector system [J].
Kinoshita, Yurie ;
Xu, Jian ;
Masuda, Akitsu ;
Minamihata, Kosuke ;
Kamiya, Noriho ;
Mon, Hiroaki ;
Fujita, Ryosuke ;
Kusakabe, Takahiro ;
Lee, Jae Man .
PROTEIN EXPRESSION AND PURIFICATION, 2019, 159 :69-74
[8]   Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor [J].
Lan, Jun ;
Ge, Jiwan ;
Yu, Jinfang ;
Shan, Sisi ;
Zhou, Huan ;
Fan, Shilong ;
Zhang, Qi ;
Shi, Xuanling ;
Wang, Qisheng ;
Zhang, Linqi ;
Wang, Xinquan .
NATURE, 2020, 581 (7807) :215-+
[9]   Efficient and cost effective production of active-form human PKB using silkworm larvae [J].
Maesaki, Ryoko ;
Satoh, Ryosuke ;
Taoka, Masato ;
Kanaba, Teppei ;
Asano, Tsunaki ;
Fujita, Chiharu ;
Fujiwara, Toshinobu ;
Ito, Yutaka ;
Isobe, Toshiaki ;
Hakoshima, Toshio ;
Maenaka, Katsumi ;
Mishima, Masaki .
SCIENTIFIC REPORTS, 2014, 4
[10]   Purification and characterization of immunogenic recombinant virus-like particles of porcine circovirus type 2 expressed in silkworm pupae [J].
Masuda, Akitsu ;
Lee, Jae Man ;
Miyata, Takeshi ;
Sato, Tetsuo ;
Hayashi, Shizuka ;
Hino, Masato ;
Morokuma, Daisuke ;
Karasaki, Noriko ;
Mon, Hiroaki ;
Kusakabe, Takahiro .
JOURNAL OF GENERAL VIROLOGY, 2018, 99 (07) :917-926