在大肠杆菌细胞中生产仅有 VP3 的腺相关病毒 2 病毒样颗粒。

IF 1.4 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2024-05-15 DOI:10.1016/j.pep.2024.106502

Chengyu Fu, Shruthi Gobbooru , Ashley T. Martino, Woon-Kai Low

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引用次数: 0

摘要

腺相关病毒（AAV）是一种前景广阔的基因治疗载体。然而，很少有研究关注在细胞中，尤其是在大肠杆菌中生产 AAV 的病毒样颗粒（VLPs）。在本研究中，我们介绍了一种通过共表达 AAV2 的 VP3 和组装激活蛋白（AAP）在大肠杆菌中生产仅有 VP3 的 AAV2 空 VLPs 的方法。虽然用我们的方法生产的 VLPs 产量较低，但这些 VLPs 能够在大肠杆菌中自我组装，无需体外胶囊组装。我们通过免疫学检测和透射电子显微镜（TEM）对所制备的 VLPs 进行了表征。总之，这项研究证明 AAV2 的囊膜可以在大肠杆菌中组装，大肠杆菌可能是生产 AAV VLPs 的候选系统。

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Production of VP3-only virus-like particles of Adeno-associated virus 2 in E. coli cells

Adeno-associated Virus (AAV) is a promising vector for gene therapy. However, few studies have focused on producing virus-like particles (VLPs) of AAV in cells, especially in E. coli. In this study, we describe a method to produce empty VP3-only VLPs of AAV2 in E. coli by co-expressing VP3 and assembly-activating protein (AAP) of AAV2. Although the yields of VLPs produced with our method were low, the VLPs were able to self-assemble in E. coli without the need of in vitro capsid assembly. The produced VLPs were characterized by immunological detection and transmission electron microscopy (TEM). In conclusion, this study demonstrated that capsid assembly of AAV2 is possible in E. coli, and E. coli may be a candidate system for production of VLPs of AAV.

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来源期刊

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and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.