当前位置: X-MOL 学术Mol. Ther. Methods Clin. Dev. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
EPPIC (Efficient Purification by Parental Inducer Constraint) Platform for Rapid Generation of Recombinant Vaccinia Viruses.
Molecular Therapy - Methods & Clinical Development ( IF 4.6 ) Pub Date : 2020-03-30 , DOI: 10.1016/j.omtm.2020.03.026
Brittany Jasperse 1 , Caitlin M O'Connell 1 , Yuxiang Wang 1 , Paulo H Verardi 1
Affiliation  

Vaccinia virus (VACV) was successfully used as a vaccine in the smallpox eradication campaign. Since then, it has been widely used in the development of vaccine and therapeutic vectors. However, methods of generating and purifying recombinant VACVs (rVACVs) are often time-consuming, cumbersome, and in some cases require specialized cell lines or equipment. Here, we describe a novel EPPIC (Efficient Purification by Parental Inducer Constraint) platform for the rapid generation of rVACVs using a replication-inducible VACV (vIND) as a parental virus for homologous recombination. Purification of the rVACV from the parental vIND is achieved by two serial passages in the absence of inducer (i.e., parental inducer "constraint") in standard laboratory cell lines, without the need for specialized equipment, within 1 week. We determined the optimal conditions for homologous recombination and serial purification and generated a suite of vIND parental viruses to facilitate customization of the platform. Importantly, the EPPIC platform can be adapted to rapidly generate replication-deficient and replication-competent rVACVs expressing vaccine or therapeutic antigens, with or without screening markers, by simple modifications to a DNA shuttle vector, thus allowing the rapid development, updating, and refinement of personalized or custom vaccines and therapeutic vectors in a matter of days.

中文翻译:

EPPIC(通过亲本诱导物约束有效纯化)平台,可快速产生重组痘苗病毒。

牛痘病毒(VACV)已成功用作根除天花的疫苗。从那时起,它已被广泛用于疫苗和治疗载体的开发。但是,产生和纯化重组VACV(rVACV)的方法通常很耗时,麻烦,并且在某些情况下需要专门的细胞系或设备。在这里,我们描述了一种新型的EPPIC(通过亲本诱导因子有效纯化)平台,用于使用复制诱导型VACV(vIND)作为亲本重组的亲本病毒快速生成rVACV。通过在标准实验室细胞系中不存在诱导剂(即,父母诱导剂“约束”)的情况下,通过两次连续传代从亲本vIND中纯化rVACV,无需专用设备,即可在1周内完成。我们确定了同源重组和系列纯化的最佳条件,并生成了一套vIND亲本病毒,以促进平台的定制。重要的是,通过对DNA穿梭载体的简单修饰,EPPIC平台可以适应快速生成表达疫苗或治疗性抗原的复制缺陷型和具有复制能力的rVACV,无论是否带有筛选标记,从而可以快速开发,更新和完善数天之内即可完成个性化或定制疫苗和治疗载体的生产。
更新日期:2020-03-30
down
wechat
bug