title: Purification and characterization of human IL-10/Fc fusion protein expressed in Pichia pastoris.
abstract: 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 5mg/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 30h 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.
authors.affiliation: Hefei National Laboratory for Physical Sciences at Microscale, Hefei, China.
authors.forename: Yugang
authors.initials: Y
authors.lastname: Guo
authors.forename: Wenyao
authors.initials: W
authors.lastname: Kang
authors.forename: Yongjun
authors.initials: Y
authors.lastname: Zhong
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.forename: Guangwei
authors.initials: G
authors.lastname: Li
authors.forename: Yi
authors.initials: Y
authors.lastname: Shen
authors.forename: Siyi
authors.initials: S
authors.lastname: Hu
authors.forename: Jie
authors.initials: J
authors.lastname: Sun
authors.forename: Weihua
authors.initials: W
authors.lastname: Xiao
mesh_terms: D000818:Animals; D006801:Humans; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D007371:Interferon-gamma / Q000032:analysis / Q000378:metabolism; D016753:Interleukin-10 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism / Q000493:pharmacokinetics; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism / Q000493:pharmacokinetics
chemical_list: C508609:IL10 protein, human; D007141:Immunoglobulin Fc Fragments; D011993:Recombinant Fusion Proteins; D016753:Interleukin-10; D007371:Interferon-gamma