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编码血液凝固因子VIII的基因表达没有B域在E. 大肠杆菌的细菌表达系统

Anna Mazurkiewicz-Pisarek¹, Alina Mazurkiewicz¹, Diana Mikiewicz²

¹Centre for Advanced Materials and Technologies, Warsaw University of Technology, Warszawa, Poland.

|October 18, 2023

概括

研究人员使用大肠杆菌系统开发了活性复合凝血因子VIII. 与传统的真核生物系统相比,这为血友病A治疗提供了潜在的更便宜和更快的方法.

科学领域:

生物技术是生物技术.
分子生物学分子生物学
生物化学生物化学

背景情况:

血液凝固因子VIII (FVIII) 对于血液静止至关重要,其缺乏导致血友病A.
目前的重组FVIII生产依赖于昂贵和耗时的真核生物系统.
细菌表达系统为重组蛋白质生产提供了一种经济有效和快速的替代方案.

研究的目的:

为了证明使用*E. coli*细菌表达系统产生活性复合FVIII的可行性.
探索一种用于FVIII生产的新型 prokaryotic 方法,作为潜在的血友病A疗法.

主要方法:

用合成FVIII基因构建一个 prokaryotic表达向量.
大肠杆菌的转化与表达载体.
优化细菌生长条件和蛋白质表达.
开发FVIII的隔离,变质和重新变质协议.
通过质谱测量确认蛋白质表达,并评估FVIII活性.

主要成果:

在大肠杆菌中成功合成和表达FVIII.
使用质谱学确认蛋白质表达.
建立了FVIII隔离和重新折叠的条件,从而产生活性蛋白.
证明了四分之一技术规模生产的可行性.

结论:

*大肠杆菌*细菌表达系统是生产活性复合FVIII的一种可行的方法.
这种方法在FVIII生产方面比传统的真核生物系统有了显著的进步.
开发的方法有望为更容易获得和更具成本效益的血友病A治疗提供希望.

关键词:

美国大肠杆菌 (E. coli).第八个因子的因素.血友病A型 A型血友病原生体表达系统的表达系统.再组合凝血因子VIII的使用.重组蛋白质生产系统的重组蛋白质生产系统.

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