Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, 8000 Utopia Parkway, Queens, NY, 11439, USA.
Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, 8000 Utopia Parkway, Queens, NY, 11439, USA.
Protein Expr Purif. 2024 Aug;220:106502. doi: 10.1016/j.pep.2024.106502. Epub 2024 May 15.
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.
腺相关病毒 (AAV) 是一种很有前途的基因治疗载体。然而,很少有研究集中在细胞中生产 AAV 的病毒样颗粒 (VLPs),特别是在大肠杆菌中。在这项研究中,我们描述了一种通过共表达 AAV2 的 VP3 和组装激活蛋白 (AAP) 在大肠杆菌中生产空 VP3 衣壳 VLPs 的方法。尽管我们的方法产生的 VLPs 产量较低,但 VLPs 能够在没有体外衣壳组装的情况下在大肠杆菌中自我组装。通过免疫检测和透射电子显微镜 (TEM) 对产生的 VLPs 进行了表征。总之,本研究表明 AAV2 的衣壳组装在大肠杆菌中是可行的,大肠杆菌可能是 AAV VLPs 的候选生产系统。