Efficient production of recombinant SARS-CoV-2 spike protein using the baculovirus-silkworm system
Efficient production of recombinant SARS-CoV-2 spike protein using the baculovirus-silkworm system
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DOI：
10.1016/j.bbrc.2020.06.020
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发表时间：
2020-08-20
影响因子：
3.1
通讯作者：
Kusakabe, Takahiro
中科院分区：
文献类型：
--
作者：
Fujita, Ryosuke;Hino, Masato;Kusakabe, Takahiro
在新的病毒性疾病爆发的情况下，需要立即开发病毒检测试剂盒和疫苗。针对COVID-19，我们利用杆状病毒-家蚕表达系统建立了一种快速生产SARS-CoV-2刺突蛋白（S蛋白）的方法。杆状病毒载体衍生的S蛋白被成功地分泌到家蚕血清中，而那些在幼虫脂肪体和蛹细胞中形成不溶性结构。具有天然序列的S蛋白的胞外域被宿主弗林蛋白酶切割，导致较少的重组蛋白产生。经弗林蛋白酶靶位点修饰的S蛋白可高效分泌到家蚕血清中，并纯化为寡聚体，其可与SARS-CoV-2 S2抗体发生免疫反应。利用重组杆状病毒直接转染家蚕，实现了无胎牛血清（FBS）系统中SARS-CoV-2S蛋白的高效表达。所得纯化的S蛋白将是用于开发免疫检测试剂盒、用于免疫球蛋白生产的免疫抗原和疫苗的有用工具。(C)2020由Elsevier Inc.出版。
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.