Immunology Center, Adolfo Lutz Institute, São Paulo 01246-902, Brazil.
Interunits Graduate Program in Biotechnology, University of São Paulo, São Paulo 05508-000, Brazil.
Int J Mol Sci. 2023 Sep 11;24(18):13955. doi: 10.3390/ijms241813955.
The Zika Virus (ZIKV) is an emerging arbovirus of great public health concern, particularly in the Americas after its last outbreak in 2015. There are still major challenges regarding disease control, and there is no ZIKV vaccine currently approved for human use. Among many different vaccine platforms currently under study, the recombinant envelope protein from Zika Virus (rEZIKV) constitutes an alternative option for vaccine development and has great potential for monitoring ZIKV infection and antibody response. This study describes a method to obtain a bioactive and functional rEZIKV using an expression system, with the aid of a 5-L airlift bioreactor and following an automated fast protein liquid chromatography (FPLC) protocol, capable of obtaining high yields of approximately 20 mg of recombinant protein per liter of bacterium cultures. The purified rEZIKV presented preserved antigenicity and immunogenicity. Our results show that the use of an airlift bioreactor for the production of rEZIKV is ideal for establishing protocols and further research on ZIKV vaccines bioprocess, representing a promising system for the production of a ZIKV envelope recombinant protein-based vaccine candidate.
Zika 病毒(ZIKV)是一种新兴的虫媒病毒,对公共卫生具有重大影响,尤其是在 2015 年最后一次爆发后在美洲。在疾病控制方面仍然存在重大挑战,目前尚无 ZIKV 疫苗获得人类使用许可。在目前正在研究的许多不同疫苗平台中,Zika 病毒重组包膜蛋白(rEZIKV)是疫苗开发的另一种选择,对于监测 ZIKV 感染和抗体反应具有巨大潜力。本研究描述了一种使用表达系统获得生物活性和功能性 rEZIKV 的方法,借助 5 升气升式生物反应器和自动化快速蛋白液相色谱(FPLC)方案,能够从每升细菌培养物中获得约 20 毫克的高产量重组蛋白。纯化的 rEZIKV 保持了抗原性和免疫原性。我们的结果表明,使用气升式生物反应器生产 rEZIKV 非常适合建立 ZIKV 疫苗生物工艺的方案和进一步研究,是生产基于 ZIKV 包膜重组蛋白的候选疫苗的有前途的系统。