title: [Expression and purification of recombinant human interleukin-11 in Pichia pastoris].
abstract: OBJECTIVE
  To express recombinant human interleukin-11 (rhIL-11) in methylotropic yeast Pichia pastoris.
  
  
  METHODS
  By designing and synthesizing an artificial gene for IL-11, the expression vector pPICZ alpha-A-IL-11 was constructed and introduced into Pichia pastoris by linearized electroporation. The rhIL-11 protein was identified by ELISA and SDS-PAGE analysis. The bioactivity was analyzed by B9-11 cell line. A combination of liquid chromatography was developed to purify the rhIL-11 from ferment supernatant.
  
  
  RESULTS
  The nucleotide sequence analysis indicated that the sequence of cloned artificial IL-11 gene accorded with that of designed; the secreted yield of rhIL-11 by yeast Pichia pastoris KM71-2424 in flask reached 60 mg/L. The biological activity of IL-11 in yeast supernatant and E. coli standard determined by B9-11 was 5.5 x 10(7) U/mg and 2.2 x 10(7) U/mg respectively. The rhIL-11 was purified to electrophoretic purity by a combination of liquid chromatography.
  
  
  CONCLUSIONS
  The human IL-11 artificial gene was obtained and successfully expressed in the Pichia pastoris(KM71-2424). The biological activity of IL-11 in yeast supernatant was significantly higher than that of E. coli standard. The rhIL-11 was purified to electrophoretic purity.
authors.affiliation: Department of Immunology, Suzhou University, Suzhou 215007, China.
authors.forename: J K
authors.initials: JK
authors.lastname: Zhu
authors.forename: Z X
authors.initials: ZX
authors.lastname: Xu
authors.forename: W D
authors.initials: WD
authors.lastname: Huang
authors.forename: T S
authors.initials: TS
authors.lastname: Ming
authors.forename: W
authors.initials: W
authors.lastname: Xie
authors.forename: Y
authors.initials: Y
authors.lastname: Xu
authors.forename: X G
authors.initials: XG
authors.lastname: Zhang
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors; D006801:Humans; D017370:Interleukin-11 / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D017370:Interleukin-11; D011994:Recombinant Proteins