CRISPR-Editing of the Vero Cell Line Improves Processability of Live Virus Vaccines by Enabling Targeted Proteolysis of Fibronectin.

IF 3.5 2区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology and Bioengineering Pub Date : 2025-05-14 DOI:10.1002/bit.29028

Jackson K B Cahn,Henry Ludwicki,Jillian Shingler,Shannon Gulvin,Young Zhang,Adam Kristopeit,Christopher Ton,Michael A Winters,James M Wagner,Jeffrey Moore

引用次数: 0

Abstract

Removal of host cell components is a significant cost driver in the production of live virus vaccines. Filtration processes such as tangential flow filtration can be effective in this capacity by leveraging the relative size difference between viral particles and host proteins; however, filtration membranes can be fouled by larger proteins, particularly those of the extracellular matrix. In this study, we used CRISPR editing to insert the recognition element of the highly-selective TEV protease into various positions of the gene encoding fibronectin in the genome of the Vero cell line, a common platform for viral production. By screening edited cell lines, we identified a promising candidate line in which fibronectin could be effectively removed by treating with the protease during processing, eliminating filter fouling and allowing for viral purification without the need for costly chromatography steps.

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crispr编辑的Vero细胞系通过激活纤维连接蛋白的靶向蛋白水解提高了活病毒疫苗的加工能力。

去除宿主细胞成分是生产活病毒疫苗的一个重要成本驱动因素。通过利用病毒颗粒和宿主蛋白之间的相对大小差异，切向流过滤等过滤过程可以有效地发挥这种能力；然而，过滤膜可能被较大的蛋白质污染，特别是那些细胞外基质。在这项研究中，我们使用CRISPR编辑将高选择性TEV蛋白酶的识别元件插入到Vero细胞系基因组中编码纤维连接蛋白的基因的各个位置，Vero细胞系是病毒产生的常见平台。通过筛选编辑细胞系，我们确定了一个有希望的候选细胞系，在加工过程中通过蛋白酶处理可以有效地去除纤维连接蛋白，消除过滤器污垢，并允许病毒纯化，而不需要昂贵的色谱步骤。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology and Bioengineering 工程技术-生物工程与应用微生物

CiteScore

7.90

自引率

5.30%

发文量

280

审稿时长

2.1 months

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