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A plant-based transient expression system for the rapid production of highly immunogenic Hepatitis E virus-like particles
Biotechnology Letters ( IF 2.0 ) Pub Date : 2020-09-01 , DOI: 10.1007/s10529-020-02995-x
Eugenia S Mardanova 1 , Katerina H Takova 2 , Valentina T Toneva 2, 3 , Gergana G Zahmanova 2, 4 , Liudmila M Tsybalova 5 , Nikolai V Ravin 1
Affiliation  

Objective Hepatitis E virus (HEV) infection is a major cause of acute hepatitis worldwide. The aim of the study is the development of plant expression system for the production of virus-like particles formed by HEV capsid and the characterization of their immunogenicity. Results Open reading frame (ORF) 2 encodes the viral capsid protein and possesses candidate for vaccine production. In this study, we used truncated genotype 3 HEV ORF 2 consisting of aa residues 110 to 610. The recombinant protein was expressed in Nicotiana benthamiana plants using the self-replicating potato virus X-based vector pEff up to 10% of the soluble protein fraction. The yield of HEV 110-610 after purification was 150–200 µg per 1 g of green leaf biomass. The recombinant protein formed nanosized virus-like particles. The immunization of mice with plant-produced HEV 110-610 protein induced high levels of HEV-specific serum antibodies. Conclusions HEV ORF 2 (110-610 aa) can be used as candidate for the development of a plant-produced vaccine against Hepatitis E.

中文翻译:


一种基于植物的瞬时表达系统,用于快速生产高免疫原性戊型肝炎病毒样颗粒



目的 戊型肝炎病毒(HEV)感染是全世界急性肝炎的主要原因。本研究的目的是开发植物表达系统,用于生产由 HEV 衣壳形成的病毒样颗粒并表征其免疫原性。结果开放阅读框(ORF)2编码病毒衣壳蛋白,具有用于疫苗生产的候选蛋白。在本研究中,我们使用由氨基酸残基 110 至 610 组成的截短基因型 3 HEV ORF 2。使用基于自我复制马铃薯病毒 X 的载体 pEff 在本塞姆氏烟草植物中表达重组蛋白,其可溶性蛋白部分的含量高达 10% 。纯化后,HEV 110-610 的产量为每 1 g 绿叶生物量 150–200 µg。重组蛋白形成纳米级病毒样颗粒。用植物产生的 HEV 110-610 蛋白免疫小鼠可诱导高水平的 HEV 特异性血清抗体。结论 HEV ORF 2 (110-610 aa) 可作为开发植物生产的戊型肝炎疫苗的候选者。
更新日期:2020-09-01
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