Expression and Purification of Bioactive High-Purity Recombinant Mouse CXCL14 in Escherichia coli

Abstract

Mouse CXCL14/BRAK is a monocyte-selective chemokine which is expressed in almost all normal tissues. A flood of reports on its new functions of tumor suppression and fat metabolism modulation has left CXCL14 a potential therapeutic candidate for these diseases. Therefore, a simple accessible method is on demand for large-scale production of recombinant mouse CXCL14 protein for in vivo animal studies. Here, we introduce an efficient method for large-scale production of recombinant mouse CXCL14, by which an 18-mg protein is produced from 2-L Escherichia coli culture with good bioactivity and low level of endotoxin.

Biophysical characterization of the isolated C‐terminal region of the transient receptor potential vanilloid 1

“…Several pieces of evidence suggest that this is not the case. Firstly, a similar refolding protocol has been used by others [35,36] and for ourselves (JLN, JB, AC, unpublished results) to refold several proteins (which also accumulate as IBs), and the refolded proteins were biologically active. Second, there are several evidences in the literature where the unfolded predictors predict a folded conformation ( Fig.…”

The Denaturing and Renaturing are Critical Steps in the Purification of Recombinant Protein in Prokaryotic System

“…However, some other proteins, such as recombinant human Midkine (rhMK), 8 recombinant mouse monokine induced by IFN-γ (rmMig) 9 , recombinant human Reg 4 (rhReg-4) 10 or recombinant human Chemerin (rhChemerin), 11 easily form inclusion bodies in E. coli, hence, necessary denaturing and renaturing steps must be applied. In some other cases, like recombinant murine CXCL14 (rmCXCL14), 12 full length protein is hard to be purified, and an additional tag will help to get final products with high yield and high purity.…”