基于重组杆状病毒表达系统生产的 BVDV E2 蛋白作为诊断抗原和免疫原的评估

IF 1.4 4区 生物学 Q4 BIOCHEMICAL RESEARCH METHODS Protein expression and purification Pub Date : 2024-09-22 DOI:10.1016/j.pep.2024.106611
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引用次数: 0

摘要

牛病毒性腹泻病毒(BVDV)是一种对全球养牛业有重大影响的重要免疫抑制病原体。目前,研究工作主要集中在 BVDV 的包膜糖蛋白 E2 上,以改善免疫反应。然而,全长 E2 蛋白作为免疫抗原和诊断工具并不理想,因此需要探索替代策略。在本研究中,我们使用 IDEB 和 ExpOptimizer 软件优化了 E2 基因,然后使用杆状病毒和大肠杆菌两种表达系统表达了 E2 基因。随后,我们评估了纯化的 E2 蛋白在小鼠体内的免疫原性及其在间接 ELISA 检测中的应用。我们的研究结果表明,与大肠杆菌系统相比,由杆状病毒系统产生的 Bac-E2 蛋白能诱导小鼠产生更高水平的抗体和脾脏淋巴细胞增殖。此外,与大肠杆菌表达的E2 ELISA相比,利用Bac-E2蛋白开发的间接ELISA检测法在特异性、灵敏度和准确性方面都更胜一筹。总之,我们的研究表明,通过杆状病毒表达系统生成的优化 E2 蛋白可在小鼠体内引起强大的体液和细胞免疫反应,因此有望成为疫苗开发的候选物质。此外,优化后的 E2 蛋白 ELISA 检测法显示出更高的灵敏度和准确性,这表明它有潜力成为一种有价值的诊断抗原。
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Evaluation of BVDV E2 proteins based on recombinant baculovirus expression system production as diagnostic antigens and immunogens
Bovine viral diarrhea virus (BVDV) is a significant immunosuppressive pathogen that has a major impact on the global cattle industry. Research efforts are currently focused on the envelope glycoprotein E2 of BVDV to improve immune responses. However, the full-length E2 protein is not ideal as an immune antigen and diagnostic tool, leading to the exploration of alternative strategies. In this study, we optimized the E2 gene using IDEB and ExpOptimizer software, then expressed the E2 gene using both baculovirus and E. coli expression systems. Subsequently, we assessed the immunogenicity of the purified E2 protein in mice and its application in indirect ELISA assays. Our findings showed that the Bac-E2 protein produced by the baculovirus system induced higher levels of antibody production and splenic lymphocyte proliferation in mice compared to the E. coli system. Moreover, the indirect ELISA assay developed using Bac-E2 protein exhibited superior specificity, sensitivity, and accuracy in comparison to the E. coli-expressed E2 ELISA. Overall, our study demonstrates that the optimized E2 protein generated through a baculovirus expression system elicits robust humoral and cellular immune responses in mice, making it a promising candidate for vaccine development. Furthermore, the optimized E2 protein ELISA assay shows enhanced sensitivity and accuracy, suggesting its potential as a valuable diagnostic antigen.
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来源期刊
Protein expression and purification
Protein expression and purification 生物-生化研究方法
CiteScore
3.70
自引率
6.20%
发文量
120
审稿时长
32 days
期刊介绍: Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.
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