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Fc-X融合蛋白在哺乳动物细胞中的高水平表达与分泌。

High level expression and secretion of Fc-X fusion proteins in mammalian cells.

作者信息

Lo K M, Sudo Y, Chen J, Li Y, Lan Y, Kong S M, Chen L, An Q, Gillies S D

机构信息

Lexigen Pharmaceuticals Corporation, Lexington, MA 02173, USA.

出版信息

Protein Eng. 1998 Jun;11(6):495-500. doi: 10.1093/protein/11.6.495.

Abstract

We have developed a general expression system that enhances the production and secretion of proteins in mammalian cells. The protein of interest is expressed as a fusion to a signal peptide and the Fc fragment of immunoglobulin as the N-terminal fusion partner, which can direct the cellular processes into expressing and secreting high levels of many different types of proteins. These include secretory proteins, enzymes and soluble domains of membrane proteins, as well as nuclear and regulatory proteins. Typical expression levels of these proteins from stable cell lines ranged from several to 100 microg/ml in conditioned media. The Fc domain helps to solubilize hydrophobic proteins and provides a handle for easy detection and purification of the fusion proteins; and it can be cleaved off by treatment with protease if desired.

摘要

我们开发了一种通用表达系统,可增强哺乳动物细胞中蛋白质的产生和分泌。目的蛋白作为与信号肽和免疫球蛋白Fc片段的融合蛋白表达,作为N端融合伴侣,它可引导细胞过程表达和分泌高水平的多种不同类型蛋白质。这些蛋白质包括分泌蛋白、酶和膜蛋白的可溶性结构域,以及核蛋白和调节蛋白。来自稳定细胞系的这些蛋白质在条件培养基中的典型表达水平为每毫升几微克至100微克。Fc结构域有助于使疏水蛋白溶解,并为融合蛋白的易于检测和纯化提供一种手段;如果需要,可用蛋白酶处理将其切除。

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