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脂多糖对人胚肾(HEK - 293)细胞培养及重组蛋白表达的影响。

Impact of lipopolysaccharides on cultivation and recombinant protein expression in human embryonal kidney (HEK-293) cells.

作者信息

Faust Christine, Beil Christian, Dittrich Werner, Rao Ercole, Langer Thomas

机构信息

R&D Biologics Research, Building H811 Sanofi-Aventis Deutschland GmbH Frankfurt am Main Germany.

出版信息

Eng Life Sci. 2021 Oct 25;21(11):778-785. doi: 10.1002/elsc.202100065. eCollection 2021 Nov.

Abstract

The human embryonal kidney 293 cell (HEK-293) is a widely used expression host for transient gene expression. The genes or plasmids used for the transient transfections are usually propagated and extracted from the gram-negative bacterium , the workhorse for molecular biologists. As a gram-negative bacterium has an outer membrane (OM) containing lipopolysaccharides (LPS) or endotoxins. LPS are very potent inducers of inflammatory cytokines in the body. In early research phases DNA intended for transient transfections is not routinely checked for LPS-levels. In this study we addressed the question whether LPS has an impact on the cultivation and production of a recombinant antibody. At high concentrations the presence of LPS has a detrimental impact on cell viability and recombinant protein expression. But low LPS concentrations are tolerated and might even enhance protein expression levels.

摘要

人胚肾293细胞(HEK - 293)是一种广泛用于瞬时基因表达的表达宿主。用于瞬时转染的基因或质粒通常从革兰氏阴性细菌中繁殖和提取,革兰氏阴性细菌是分子生物学家的得力工具。作为革兰氏阴性细菌,其具有包含脂多糖(LPS)或内毒素的外膜(OM)。LPS是体内炎症细胞因子的非常有效的诱导剂。在早期研究阶段,用于瞬时转染的DNA通常不会常规检查LPS水平。在本研究中,我们探讨了LPS是否对重组抗体的培养和生产有影响这一问题。在高浓度下,LPS的存在对细胞活力和重组蛋白表达有不利影响。但低浓度的LPS是可以耐受的,甚至可能提高蛋白表达水平。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d1c3/8576075/735f963af7d3/ELSC-21-778-g004.jpg

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