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基于豇豆花叶病毒外壳蛋白的嵌合 SARS-CoV-2 抗原的制备与鉴定。

Production and characterization of chimeric SARS-CoV-2 antigens based on the capsid protein of cowpea chlorotic mottle virus.

机构信息

Facultad de Ciencias Químicas, Universidad Autónoma de San Luis Potosí, Av. Dr. Manuel Nava 6, SLP 78210, Mexico; Sección de Biotecnología, Centro de Investigación en Ciencias de la Salud y Biomedicina, Universidad Autónoma de San Luis Potosí, Av. Sierra Leona 550, Lomas 2ª. Sección, San Luis Potosí 78210, Mexico.

Sección de Biotecnología, Centro de Investigación en Ciencias de la Salud y Biomedicina, Universidad Autónoma de San Luis Potosí, Av. Sierra Leona 550, Lomas 2ª. Sección, San Luis Potosí 78210, Mexico.

出版信息

Int J Biol Macromol. 2022 Jul 31;213:1007-1017. doi: 10.1016/j.ijbiomac.2022.06.021. Epub 2022 Jun 8.

Abstract

The COVID-19 pandemic has highlighted the need for new vaccine platforms to rapidly develop solutions against emerging pathogens. In particular, some plant viruses offer several advantages for developing subunit vaccines, such as high expression rates in E. coli, high immunogenicity and safety, and absence of pre-immunity that could interfere with the vaccine's efficacy. Cowpea chlorotic mottle virus (CCMV) is a model system that has been extensively characterized, with key advantages for its use as an epitope carrier. In the present study, three relevant epitopes from the SARS-CoV-2 Spike protein were genetically inserted into the CCMV CP and expressed in E. coli cultures, resulting in the CCMV1, CCMV2, and CCMV3 chimeras. The recombinant CP mutants were purified from the formed inclusion bodies and refolded, and their immunogenicity as a subunit vaccine was assessed in BALB/c mice. The three mutants are immunogenic as they induce high IgG antibody titers that recognize the recombinant full-length S protein. This study supports the application of CCMV CP as an attractive carrier for the clinical evaluation of vaccine candidates against SARS-CoV-2. Furthermore, it suggests that VLPs assembled from these chimeric proteins could result in antigens with better immunogenicity.

摘要

COVID-19 大流行凸显了需要新的疫苗平台来快速针对新出现的病原体开发解决方案。特别是,一些植物病毒为开发亚单位疫苗提供了几个优势,例如在大肠杆菌中的高表达率、高免疫原性和安全性,以及不存在可能干扰疫苗效力的预先免疫。豇豆花叶病毒(CCMV)是一个经过广泛表征的模型系统,具有作为表位载体使用的关键优势。在本研究中,三个来自 SARS-CoV-2 刺突蛋白的相关表位被遗传插入到 CCMV CP 中,并在大肠杆菌培养物中表达,产生了 CCMV1、CCMV2 和 CCMV3 嵌合体。重组 CP 突变体从形成的包涵体中纯化并复性,并在 BALB/c 小鼠中评估其作为亚单位疫苗的免疫原性。这三种突变体具有免疫原性,因为它们诱导了高 IgG 抗体滴度,可识别重组全长 S 蛋白。本研究支持将 CCMV CP 作为 SARS-CoV-2 候选疫苗临床评估的有吸引力的载体的应用。此外,它表明由这些嵌合蛋白组装的 VLPs 可能产生具有更好免疫原性的抗原。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a14/9174154/0145b9746e4a/gr1_lrg.jpg

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