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用于生产重组蛋白(包括人源化抗V5标签抗体)的单周期5型副流感病毒载体

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

作者信息

Randall Richard E, Young Dan, Pisliakova Maria, Andrejeva Jelena, West Lynsey, Rossler Luis, Morath Volker, Hughes David, Goodbourn Steve

机构信息

School of Biology, Centre for Biomolecular Sciences, BMS Building, North Haugh, University of St. Andrews, St. Andrews, Fife, KY16 9ST, UK.

Department of Nuclear Medicine, University Clinic rechts der Isar, School of Medicine, Technical University of Munich, 81675 Munich, Germany.

出版信息

J Gen Virol. 2025 Jan;106(1). doi: 10.1099/jgv.0.002061.

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 after 5 days when cells were grown to a density of ~1×10 cells per millilitre at the time of infection. We suggest that PIV5-based vectors may be further developed to produce recombinant proteins both and .

摘要

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/裂解性感染。在此,我们构建了表达PIV5融合蛋白(F)的辅助细胞系,该细胞系可支持缺失F基因的病毒的复制。作为缺失F基因的单周期感染性病毒可作为安全高效表达载体这一原理的证据,我们克隆并表达了小鼠抗V5标签抗体(克隆号SV5-Pk1)的人源化(Hu)版本。我们发现,多种不同的细胞系均可被感染并高水平表达Hu抗V5抗体,当感染时细胞密度达到约每毫升1×10个细胞,中国仓鼠卵巢细胞在5天后可表达20 - 50毫克/升。我们认为,基于PIV5的载体可能会得到进一步开发,用于在[此处原文缺失具体内容]和[此处原文缺失具体内容]中生产重组蛋白。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f769/12282297/41886b501447/jgv-106-02061-g001.jpg

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