丙型肝炎病毒核心-NS3融合蛋白在细菌粪便表面的表达:疫苗生产前景。

Q3 Biochemistry, Genetics and Molecular Biology Avicenna journal of medical biotechnology Pub Date : 2023-07-01 DOI:10.18502/ajmb.v15i3.12927
Minoosadat Tayebinia, Sedigheh Sharifzadeh, Gholamreza Rafiei Dehbidi, Farahnaz Zare, Reza Ranjbaran, Amir Rahimi, Mohammad Reza Miri, Mehdi Mirzakhani, Abbas Behzad-Behbahania
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摘要

背景:一方面,使用细菌表面显示系统的抗原呈递具有细菌载体的优点,包括低成本生产和易于操作。另一方面,细菌可以作为佐剂帮助刺激免疫系统。例如,利用细菌表面显示技术,我们开发了一种丙型肝炎病毒(HCV)多重抗原,显示细菌表面,然后将其变成细菌幽灵。方法:将HCV核心和NS3蛋白的保守表位克隆到AIDA基因质粒中,作为自身转运蛋白。然后将重组质粒转化到大肠杆菌(E.coli)Bl21(DE3)中。然后,重组细菌变成了一个细菌幽灵,一个空的细胞包膜。采用全细胞ELISA、流式细胞仪和蛋白质印迹技术监测细菌表面蛋白质的表达。结果:HCV核心-NS3-AIDA融合蛋白在大肠杆菌Bl21(DE3)表面成功表达,并通过蛋白质印迹、酶联免疫吸附试验(ELISA)和流式细胞仪检测技术进行了证实。结论:HCV抗原在非致病菌表面的存在有望开发出安全、成本效益高的疫苗,具有最佳的内在佐剂效果和异源抗原暴露于免疫系统。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Expression of the Hepatitis C Virus core-NS3 Fusion Protein on the Surface of Bacterial Ghosts: Prospects for Vaccine Production.

Background: Antigen presentation using bacterial surface display systems, on one hand, has the benefits of bacterial carriers, including low-cost production and ease of manipulation. On the other hand, the bacteria can help in stimulating the immune system as an adjuvant. For example, using bacterial surface display technology, we developed a hepatitis C virus (HCV) multiple antigens displaying bacteria's surface and then turned it into a bacterial ghost.

Methods: The HCV core and NS3 proteins' conserved epitopes were cloned into the AIDA gene plasmid as an auto transporter. The recombinant plasmid was then transformed into Escherichia coli (E. coli) Bl21 (DE3). Recombinant bacteria were then turned into a bacterial ghost, an empty cell envelope. Whole-cell ELISA, flow cytometry, and Western blot techniques were used for monitoring the expression of proteins on the surface of bacteria.

Results: A fusion protein of HCV core-NS3-AIDA was successfully expressed on the E. coli Bl21 (DE3) surface and confirmed by western blotting, Enzyme-Linked Immunosorbent Assay (ELISA), and flow cytometry detection techniques.

Conclusion: The presence of HCV antigens on non-pathogen bacteria surfaces holds promise for developing safe and cost-benefit-accessible vaccines with optimal intrinsic adjuvant effects and exposure of heterologous antigens to the immune system.

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来源期刊
Avicenna journal of medical biotechnology
Avicenna journal of medical biotechnology Biochemistry, Genetics and Molecular Biology-Biotechnology
CiteScore
2.90
自引率
0.00%
发文量
43
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