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腺病毒载体的可扩展生产。

Scalable production of adenovirus vectors.

作者信息

Silva Ana Carina, Fernandes Paulo, Sousa Marcos F Q, Alves Paula M

机构信息

Animal Cell Technology Unit, Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa (ITQB-UNL), Oeiras, Portugal.

出版信息

Methods Mol Biol. 2014;1089:175-96. doi: 10.1007/978-1-62703-679-5_13.

Abstract

Recombinant adenoviruses (AdV) are highly efficient at gene transfer for a broad spectrum of cell types and species. They became one of the vectors of choice for gene delivery and expression of foreign proteins in gene therapy and vaccination purposes. To meet the need of significant amounts of adenoviral vectors for preclinical and possibly clinical uses, scalable and reproducible production processes are required.In this chapter, we review processes used for scalable production of two types of first generation (E1-deleted) adenoviral vectors (Human and Canine) using stirred tank bioreactors. The production of adenovirus vectors using either suspension (HEK 293) or anchorage-dependent cells (MDCK-E1) are described to exemplify scalable production processes with different cell-culture types. The downstream processes will be covered in the next chapter.

摘要

重组腺病毒(AdV)在将基因转移到广泛的细胞类型和物种方面效率很高。它们成为基因治疗和疫苗接种中基因递送和外源蛋白表达的首选载体之一。为满足临床前和可能临床应用中大量腺病毒载体的需求,需要可扩展且可重复的生产工艺。在本章中,我们回顾了使用搅拌罐生物反应器可扩展生产两种第一代(E1缺失)腺病毒载体(人源和犬源)的工艺。描述了使用悬浮细胞(HEK 293)或贴壁依赖性细胞(MDCK-E1)生产腺病毒载体的过程,以举例说明不同细胞培养类型的可扩展生产工艺。下游工艺将在下一章介绍。

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