从COS-1细胞中生产和纯化登革病毒样颗粒
Production and Purification of Dengue Virus-like Particles from COS-1 Cells.
作者信息
Galula Jedhan Ucat, Chang Gwong-Jen J, Chao Day-Yu
机构信息
Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung Hsing University, Taichung, Taiwan.
Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, US Department of Health and Human Services, Fort Collins, Colorado, USA.
出版信息
Bio Protoc. 2019 Jun 20;9(12):e3280. doi: 10.21769/BioProtoc.3280.
Non-infectious virus-like particles (VLPs) containing dengue virus (DENV) pre-membrane (prM) and envelope (E) proteins have been demonstrated to be highly immunogenic and can be used as a potential vaccine candidate as well as a tool for serodiagnostic assays. Successful application of VLPs requires abundant, and high-purity production methods. Here, we describe a robust protocol for producing DENV VLPs from transiently-transformed or stable COS-1 cells and further provide an easily adaptable antigen purification method by sucrose gradient centrifugation.
含有登革病毒（DENV）前膜（prM）和包膜（E）蛋白的非感染性病毒样颗粒（VLP）已被证明具有高度免疫原性，可作为潜在的疫苗候选物以及血清诊断检测的工具。VLP的成功应用需要丰富且高纯度的生产方法。在此，我们描述了一种从瞬时转染或稳定的COS-1细胞中生产DENV VLP的稳健方案，并进一步提供了一种通过蔗糖梯度离心易于调整的抗原纯化方法。
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