Yield Optimisation of Hepatitis B Virus Core Particles in E. coli Expression System for Drug Delivery Applications.
Yield Optimisation of Hepatitis B Virus Core Particles in E. coli Expression System for Drug Delivery Applications.
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DOI：
10.1038/srep43160
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发表时间：
2017-03-03
影响因子：
4.6
通讯作者：
Al-Jamal KT
中科院分区：
文献类型：
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作者：
Bin Mohamed Suffian IF;Garcia-Maya M;Brown P;Bui T;Nishimura Y;Palermo AR;Ogino C;Kondo A;Al-Jamal KT
大肠大肠杆菌表达系统为快速、高产、经济地生产B型肝炎病毒核心（HBc）颗粒提供了一种方法。然而，在细菌中高水平生产HBc颗粒是需要的，并且从E.大肠杆菌中的表达，以提高实验室规模的生产力，用于进一步的药物递送应用。生产步骤包括细菌培养、蛋白质分离、变性、纯化和最后的蛋白质组装。在这项研究中，我们描述了一个修改后的E。大肠杆菌为基础的方法纯化HBc颗粒，并与使用常规纯化方法获得的结果进行比较。通过原子力显微镜（AFM）证实HBc颗粒形态。蛋白质特异性和二级结构分别通过Western Blot和圆二色性（CD）进行确认。与常规方法相比，改进的方法产生了约3倍更高的产量和更高纯度的野生型HBc颗粒。我们的结果表明，改进的方法产生了更好的收率和纯度的HBc颗粒在E。大肠杆菌表达系统，其被充分表征并适合用于药物递送应用。
An E. coli expression system offers a mean for rapid, high yield and economical production of Hepatitis B Virus core (HBc) particles. However, high-level production of HBc particles in bacteria is demanding and optimisation of HBc particle yield from E. coli is required to improve laboratory-scale productivity for further drug delivery applications. Production steps involve bacterial culture, protein isolation, denaturation, purification and finally protein assembly. In this study, we describe a modified E. coli based method for purifying HBc particles and compare the results with those obtained using a conventional purification method. HBc particle morphology was confirmed by Atomic Force Microscopy (AFM). Protein specificity and secondary structure were confirmed by Western Blot and Circular Dichroism (CD), respectively. The modified method produced ~3-fold higher yield and greater purity of wild type HBc particles than the conventional method. Our results demonstrated that the modified method produce a better yield and purity of HBc particles in an E. coli-expression system, which are fully characterised and suitable to be used for drug delivery applications.