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High level soluble expression, purification, and characterization of human ciliary neuronotrophic factor in Escherichia coli by single protein production system.

Abstract：

:Ciliary neurotrophic factor (CNTF) is characterized as a neuropoietic cytokine for a broad spectrum of neurons, leading to its evaluation in humans suffering from neurodegenerative diseases. Due to its wide range of biological applications, high yield production of soluble biologically active recombinant human CNTF (rhCNTF) in heterologous expression system is demanded. Many attempts had been undertaken to product rhCNTF in Escherichia coli (E. coli), however, the expression level of rhCNTF was low and most of which formed insoluble inclusion bodies. In this study, we described a new and efficient method to express rhCNTF. The human CNTF gene was codon optimized and then expressed by the single protein production (SPP) expression system in E. coli. The results showed that rhCNTF was expressed as a soluble biologically active protein, and upon purification, the final yield was about 250 mg/L in shake flask with a specific neuroprotective activity in Aβ-induced SH-SY5Y cell injury model. Our study might open up a new strategy for large-scale production of functional rhCNTF for clinical applications as well as basic research.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Wang K,Zhou F,Zhu L,Zhu X,Zhang K,Zhu L

doi

10.1016/j.pep.2014.01.008

subject

Has Abstract

pub_date

2014-04-01 00:00:00

pages

8-13

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(14)00010-2

journal_volume

96

pub_type

杂志文章

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