title: [Eukaryotic expression and biological activity of human interleukin-35].
abstract: OBJECTIVE
  To express and purify recombinant human interleukin-35[IL-35-IgG1 (Fc) in eukaryotic expression system and to study the interaction of IL-35 with gp130 protein.
  
  
  METHODS
  A mammalian expression vector, pSTEP2-IL35-LFc, was constructed and transfected into HEK293T cells. Then rhIL-35-IgG1 (Fc) was expressed and purified with protein A affinity chromatography, and was examined with SDS-PAGE and Western blot analysis. The binding of IL-35 to its receptor gp130 was investigated using enzyme-linked immunosorbent assay (ELISA). The biological effect of IL-35 on gp130 was explored in M1 myeloid leukemia cells.
  
  
  RESULTS
  rhIL-35-Fc with high purity on reduced SDS-PAGE was obtained. ELISA confirmed that IL-35-Fc was bound to gp130 and neutralized the function of gp130 in M1 myeloid leukemic cells.
  
  
  CONCLUSIONS
  High-purity and biologically active rhIL-35-Fc protein successfully produced in this study. IL-35 binds to gp130 and neutralizes its activity of in M1 myeloid leukemic cells.
authors.affiliation: Cell Culture R&D Center, CAMS and PUMC, Beijing 100005, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Ma
authors.affiliation: Cell Culture R&D Center, CAMS and PUMC, Beijing 100005, China; Sino Biogical Inc, Beijing 100176, China.
authors.forename: Liang-zhi
authors.initials: LZ
authors.lastname: Xie
mesh_terms: D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007378:Interleukins / Q000378:metabolism*; D014162:Transfection
chemical_list: D007378:Interleukins; C532417:interleukin-35, human