Single-cycle parainfluenza virus type 5 vectors for producing recombinant proteins, including a humanized anti-V5 tag antibody.

IF 3.6 4区 医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of General Virology Pub Date : 2025-01-01 DOI:10.1099/jgv.0.002061
Richard E Randall, Dan Young, Maria Pisliakova, Jelena Andrejeva, Lynsey West, Luis Rossler, Volker Morath, David Hughes, Steve Goodbourn
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Abstract

Parainfluenza virus type 5 (PIV5) can cause either persistent or acute/lytic infections in a wide range of mammalian tissue culture cells. Here, we have generated PIV5 fusion (F)-expressing helper cell lines that support the replication of F-deleted viruses. As proof of the principle that F-deleted single-cycle infectious viruses can be used as safe and efficient expression vectors, we have cloned and expressed a humanized (Hu) version of the mouse anti-V5 tag antibody (clone SV5-Pk1). We show that multiple different cell lines can be infected and express high levels of the Hu anti-V5 antibody, with Chinese hamster ovary cells expressing 20-50 mg l-1 after 5 days when cells were grown to a density of ~1×106 cells per millilitre at the time of infection. We suggest that PIV5-based vectors may be further developed to produce recombinant proteins both in vitro and in vivo.

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单循环副流感病毒5型载体,用于生产重组蛋白,包括人源化抗v5标签抗体。
5型副流感病毒(PIV5)可在多种哺乳动物组织培养细胞中引起持续性或急性/溶解性感染。在这里,我们生成了表达PIV5融合(F)的辅助细胞系,支持F缺失病毒的复制。为了证明f缺失的单循环感染性病毒可以作为安全高效的表达载体,我们克隆并表达了人源化(Hu)版本的小鼠抗v5标签抗体(克隆SV5-Pk1)。我们发现多种不同的细胞系可以被感染并表达高水平的Hu抗v5抗体,当细胞生长到感染时每毫升约1×106细胞的密度时,中国仓鼠卵巢细胞在5天后表达20-50 mg l-1。我们认为,基于piv5的载体可以进一步发展,在体外和体内产生重组蛋白。
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来源期刊
Journal of General Virology
Journal of General Virology 医学-病毒学
CiteScore
7.70
自引率
2.60%
发文量
91
审稿时长
3 months
期刊介绍: JOURNAL OF GENERAL VIROLOGY (JGV), a journal of the Society for General Microbiology (SGM), publishes high-calibre research papers with high production standards, giving the journal a worldwide reputation for excellence and attracting an eminent audience.
期刊最新文献
Erratum: Out-of-sync evolutionary patterns and mutual interplay of major and minor capsid proteins in norovirus GII.2. Targeting pseudoknots with Cas13b inhibits porcine epidemic diarrhoea virus replication. A yeast-assembled, plasmid-launched reverse genetics system for the murine coronavirus MHV-A59. An improved reverse genetics system for rotavirus vaccine strain LLR using five plasmid vectors. Single-cycle parainfluenza virus type 5 vectors for producing recombinant proteins, including a humanized anti-V5 tag antibody.
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