Expression, purification and characterization of recombinant human interleukin-22 in Pichia pastoris.
Interleukin-22 (IL-22) is a member of the IL-10 family. Its potential in clinical use has been highlighted for its important roles in promoting antimicrobial defense and preventing epithelial damages. Previous studies have reported that IL-22 can be expressed using prokaryotic systems and purified from inclusion bodies, however the recovery rate was poor. To produce functional IL-22 with a high yield, human IL-22 was inserted into the eukaryotic expression vector pPICZalphaA and transformed into Pichia pastoris. The expression of recombinant human IL-22 (rhIL-22) was induced by methanol and accounted for about 85% of the total secreted proteins. A simple purification strategy was established to purify the rhIL-22 from the culture supernatant, yielding 100 mg/l at 90% purity by chromatography with a SP Sepharose FF column. Bioactivity analysis showed the purified rhIL-22 demonstrated a specific activity that was comparable with the commercial one. This study provides a new strategy for large-scale production of bioactive IL-22 for use in basic studies and therapeutic applications.
Cai
Xin
X
Laboratory of Molecular Genetics, Beijing Institute of Basic Medical Sciences, 100850 Beijing, China.
Wang
Jinfeng
J
Wang
Yuanyuan
Y
Yang
Yongchang
Y
Gao
Jie
J
Fu
Wenliang
W
Wang
Jiaxi
J
Xu
Donggang
D
D001681:Biological Assay; D056945:Hep G2 Cells; D006801:Humans; D007378:Interleukins / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D010843:Pichia / Q000187:drug effects / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D014170:Transformation, Genetic / Q000187:drug effects; D000097043:Interleukin-22
D007378:Interleukins; D011994:Recombinant Proteins; D000097043:Interleukin-22