کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2020651 | 1069196 | 2012 | 5 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Purification and characterization of human IL-10/Fc fusion protein expressed in Pichia pastoris Purification and characterization of human IL-10/Fc fusion protein expressed in Pichia pastoris](/preview/png/2020651.png)
Interleukin (IL)-10 is an anti-inflammatory cytokine that could be potentially applied for clinical therapy. However, its short circulating half-life in the serum limits its clinical applications. In this study, we designed a fusion protein containing human IL-10 and an IgG Fc fragment (hIL-10/Fc), and expressed it in Pichia pastoris. This hIL-10/Fc fusion protein was purified from the culture supernatant using MabSelect affinity chromatography and size-exclusion chromatography. The hIL-10/Fc yield was about 5 mg/L in shake flasks, with purity exceeding 95%. In addition, the hIL-10/Fc fusion protein suppressed the phytohemagglutinin-induced IFN-γ production in human peripheral blood mononuclear cells. Pharmacokinetic study also revealed that hIL-10/Fc has a prolonged circulating half-life of about 30 h in rats. More importantly, the hIL-10/Fc fusion protein displayed highly specific biological activity, which was slightly higher than that of the commercial recombinant human IL-10 (rhIL-10). Therefore, P. pastoris is useful in the large-scale production of hIL-10/Fc fusion protein for both research and therapeutic applications.
► This is the first study on the expression of human IL-10/Fc in P. pastoris.
► IL-10/Fc shows an increasing bioactivity compare to rhIL-10.
► IL-10/Fc expressed in P. pastoris is in the form of dimmeric.
Journal: Protein Expression and Purification - Volume 83, Issue 2, June 2012, Pages 152–156