一种用于快速生产高免疫原性戊型肝炎病毒样颗粒的基于植物的瞬时表达系统。

A plant-based transient expression system for the rapid production of highly immunogenic Hepatitis E virus-like particles.

作者信息

Mardanova Eugenia S, Takova Katerina H, Toneva Valentina T, Zahmanova Gergana G, Tsybalova Liudmila M, Ravin Nikolai V

机构信息

Institute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, 119071, Russia.

Department of Plant Physiology and Molecular Biology, University of Plovdiv, 4000, Plovdiv, Bulgaria.

出版信息

Biotechnol Lett. 2020 Nov;42(11):2441-2446. doi: 10.1007/s10529-020-02995-x. Epub 2020 Sep 1.

DOI:10.1007/s10529-020-02995-x

PMID:32875477

Abstract

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.

摘要

目的

戊型肝炎病毒（HEV）感染是全球急性肝炎的主要病因。本研究旨在开发用于生产由HEV衣壳形成的病毒样颗粒的植物表达系统，并对其免疫原性进行表征。

结果

开放阅读框（ORF）2编码病毒衣壳蛋白，是疫苗生产的候选对象。在本研究中，我们使用了截短的3型HEV ORF 2，其由第110至610位氨基酸残基组成。重组蛋白通过基于自我复制马铃薯X病毒的载体pEff在本氏烟草植物中表达，其表达量高达可溶性蛋白组分的10%。纯化后每1克绿叶生物量中HEV 110 - 610的产量为150 - 200微克。重组蛋白形成了纳米级的病毒样颗粒。用植物产生的HEV 110 - 610蛋白免疫小鼠可诱导产生高水平的HEV特异性血清抗体。