Expression and one-step ion-exchange purification of (AAR)IL-8 (human IL-8 receptor antagonist)

Protein Expr Purif. 2005 Dec;44(2):104-9. doi: 10.1016/j.pep.2005.03.033.

Abstract

Interleukin-8 (IL-8) is C-X-C chemokine, which is produced by a variety of cells. IL-8 plays an important role in the inflammatory response and may be a therapeutic target for some inflammatory diseases. To develop an IL-8 receptor antagonist, (AAR)IL-8 (IL-8 receptor antagonist) was constructed and successfully expressed in Escherichia coli. (AAR)IL-8 could be easily purified by one-step SP-Sepharose fast flow column after the lysate of recombinant bacterial cells was heated at 70 degrees C for 10 min. The purity of (AAR)IL-8 is more than 95%. This purification process resulted in final purified yields of 4.29 mg (AAR)IL-8/g cell paste. In addition, the purified (AAR)IL-8 can significantly inhibit the chemotaxis that was induced by human IL-8 in vitro and in vivo. These results showed that this purification process is very simple and effective. It could be easily amplified at a larger scale. (AAR)IL-8 might find use as a new therapeutic IL-8 receptor antagonist for some acute and chronic inflammatory diseases.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution
  • Animals
  • Chemotaxis / drug effects
  • Escherichia coli / genetics
  • Humans
  • Interleukin-8 / biosynthesis*
  • Interleukin-8 / genetics
  • Interleukin-8 / pharmacology
  • Neutrophils / cytology
  • Neutrophils / drug effects
  • Plasmids / genetics
  • Protein Engineering / methods
  • Rats
  • Receptors, Interleukin-8A / antagonists & inhibitors*
  • Receptors, Interleukin-8B / antagonists & inhibitors*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / isolation & purification*
  • Recombinant Proteins / pharmacology

Substances

  • Interleukin-8
  • Receptors, Interleukin-8A
  • Receptors, Interleukin-8B
  • Recombinant Proteins