Expression and one-step ion-exchange purification of (AAR)IL-8 (human IL-8 receptor antagonist).
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.
Hou
Dengyong
D
Biotechnology Center, The Fourth Military Medical University, Xi'an 710032, PR China.
Yan
Zhen
Z
Shi
Jihong
J
Han
Wei
W
Zhang
Yingqi
Y
D019943:Amino Acid Substitution; D000818:Animals; D002633:Chemotaxis / Q000187:drug effects; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering / Q000379:methods; D051381:Rats; D023062:Receptors, Interleukin-8A / Q000037:antagonists & inhibitors*; D023063:Receptors, Interleukin-8B / Q000037:antagonists & inhibitors*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification* / Q000494:pharmacology
D016209:Interleukin-8; D023062:Receptors, Interleukin-8A; D023063:Receptors, Interleukin-8B; D011994:Recombinant Proteins