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用于生产口蹄疫病毒空衣壳的无血清悬浮培养哺乳动物细胞中的瞬时基因表达

Transient gene expression in serum-free suspension-growing mammalian cells for the production of foot-and-mouth disease virus empty capsids.

作者信息

Mignaqui Ana Clara, Ruiz Vanesa, Perret Sylvie, St-Laurent Gilles, Singh Chahal Parminder, Transfiguracion Julia, Sammarruco Ayelén, Gnazzo Victoria, Durocher Yves, Wigdorovitz Andrés

机构信息

Instituto de Virología, Centro de Investigación en Ciencias Veterinarias y Agronómicas, Instituto Nacional de Tecnología Agropecuaria, Hurlingham, Buenos Aires, Argentina.

出版信息

PLoS One. 2013 Aug 20;8(8):e72800. doi: 10.1371/journal.pone.0072800. eCollection 2013.

Abstract

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals. It produces severe economic losses in the livestock industry. Currently available vaccines are based on inactivated FMD virus (FMDV). The use of empty capsids as a subunit vaccine has been reported to be a promising candidate because it avoids the use of virus in the vaccine production and conserves the conformational epitopes of the virus. In this report, we explored transient gene expression (TGE) in serum-free suspension-growing mammalian cells for the production of FMDV recombinant empty capsids as a subunit vaccine. The recombinant proteins produced, assembled into empty capsids and induced protective immune response against viral challenge in mice. Furthermore, they were recognized by anti-FMDV bovine sera. By using this technology, we were able to achieve expression levels that are compatible with the development of a vaccine. Thus, TGE of mammalian cells is an easy to perform, scalable and cost-effective technology for the production of a recombinant subunit vaccine against FMDV.

摘要

口蹄疫(FMD)是偶蹄动物的一种高度传染性疾病。它给畜牧业造成严重的经济损失。目前可用的疫苗基于灭活口蹄疫病毒(FMDV)。据报道,使用空衣壳作为亚单位疫苗是一种有前景的候选疫苗,因为它避免了在疫苗生产中使用病毒,并保留了病毒的构象表位。在本报告中,我们探索了在无血清悬浮生长的哺乳动物细胞中进行瞬时基因表达(TGE),以生产FMDV重组空衣壳作为亚单位疫苗。所产生的重组蛋白组装成空衣壳,并在小鼠中诱导针对病毒攻击的保护性免疫反应。此外,它们能被抗FMDV牛血清识别。通过使用这项技术,我们能够实现与疫苗开发相匹配的表达水平。因此,哺乳动物细胞的TGE是一种易于操作、可扩展且具有成本效益的技术,用于生产抗FMDV的重组亚单位疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dd5a/3748020/b93087452770/pone.0072800.g001.jpg

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