相关实验视频
Updated: Aug 3, 2026

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The MultiBac Protein Complex Production Platform at the EMBL
Published on: July 11, 2013
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针对功能性重组SARS-CoV-2尖端S1受体结合域蛋白的优化过程的开发,该蛋白在百科病毒表达载体系统中产生
Mohamed Boumaiza1, Ameni Chaabene1, Ines Akrouti1
1Laboratory of Molecular Microbiology, Vaccinology and Biotechnology Development, Group of Biotechnology Development, Institut Pasteur de Tunis, Université Tunis El Manar, Tunis 1002, Tunisia.
Tropical medicine and infectious disease
|November 24, 2023
概括
研究人员开发了一种负担得起的方法来生产SARS-CoV-2抗原用于本地血清学测试. 这种方法提高了COVID-19监测能力,特别是在中低收入国家,使用S1RBD蛋白通过baculovirus表达.
科学领域:
- 病毒学 病毒学
- 免疫学 免疫学 免疫学
- 生物技术是生物技术.
背景情况:
- 绘制严重急性呼吸系统综合征冠状病毒2 (SARS-CoV-2) 传播和免疫反应的地图需要有效的本地血清学测试.
- SARS-CoV-2 免疫原蛋白的高成本限制了中低收入国家 (LMICs) 的可访问性.
研究的目的:
- 建立一个负担得起的生产方法,用于SARS-CoV-2受体结合域 (RBD) 的尖端S1蛋白 (S1RBD) 使用baculovirus表达载体系统 (BEVS).
- 优化S1RBD高产量的生产条件,并评估其抗原性,用于开发诊断工具.
主要方法:
- 使用BEVS.生产S1RBD.
- 优化培养条件,包括感染的多重性 (MOI),细胞密度和收获时间.
- 西方斑分析用于监测S1RBD表达和评估对COVID-19患者血清的抗原性.
主要成果:
- 确定了最佳生产条件:MOI 3,细胞密度为2-3 × 10^6细胞/毫升,感染后的收获时间为72-96小时.
- 获得了高S1RBD产量:在摇瓶中达到4mg/L,在7L生物反应器中达到70mg/L.
- 生产的S1RBD对来自COVID-19患者的血清表现出极好的抗原性.
结论:
- 该研究成功建立了一种有效和可扩展的方法来产生SARS-CoV-2 S1RBD.
- 开发的S1RBD是一种可行的抗原,用于创建成本效益的血清分析,如ELISA用于COVID-19监测.
- 这种方法可以支持本地诊断测试开发和血清流行研究,特别是在资源有限的环境中.
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