登革热病毒3型E蛋白域III的细胞外表达和诊断潜力通过Komagataella phaffii中的Codon优化
Camila Marques de Simone1, Maria Carolina Yi Lin Lee1, Yasmin Mayra Bispo1
1Laboratory of Microorganism Physiology, Department of Biomedical Sciences and Health, Universidade do Estado de Minas Gerais (UEMG), Rua Sabará, 164, Centro, Passos, MG, CEP: 37900-004, Brazil.
Molecular biotechnology
|July 3, 2025
概括
研究人员开发了一种具有成本效益的方法,使用Komagataella phaffii.产生登革热病毒EDIII抗原. 这种重组抗原显示出开发精确登革热诊断测试的前景.
科学领域:
- 病毒学 病毒学
- 生物技术是生物技术.
- 免疫学 免疫学 免疫学
背景情况:
- 登革热病毒 (DENV) 影响全球数百万人,由蚊子传播.
- DENV包膜蛋白域III (EDIII) 对于病毒进入至关重要,也是诊断和疫苗的关键目标.
研究的目的:
- 使用Komagataella phaffii酵母表达系统来表达和净化DENV-3中的EDIII蛋白.
- 在诊断试验中评估复合EDIII抗原的疗效.
主要方法:
- 在K. phaffii集成的PPICZαA载体中优化和克隆EDIII基因.
- 由甲醇诱导的蛋白质表达和通过Ni2+亲和染色法进行净化.
- 使用SDS-PAGE和西方斑点分析确认蛋白质表达.
主要成果:
- 实现了净化的重组EDIII (209-309 mg/L) 的高产量.
- 使用92个血清样本进行间接ELISA测试,显示出具有很高的诊断潜力.
- 显示IgM的AUC为0.94,IgG的AUC为0.828,IgM检测的特异性为92%.
结论:
- 科玛格塔贝拉 (Komagataella phaffii) 是一个有效的宿主,可以产生复合登革热EDIII抗原.
- 这种方法为开发登革热诊断工具提供了一个可扩展和经济的战略.
- 这种重组抗原显着有望改善登革热诊断.
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