title: [Study on the detection method for determining the bioactivity of recombinant human interleukin-22].
abstract: OBJECTIVE
  To develop a method for determining the bioactivity of recombinant human IL-22.
  
  
  METHODS
  pSTAT3-TA-Luc and pcDNA3. 1 plasmids were co-transfect to HepG2 cells to generate stable HepG2/ STAT3 cell line with G418 screen. After treating cell with IL-22, the luciferase activity was assayed. Orthogonal test was used to optimize the assay condition, and then the reproducibility and specificity of the assay was checked.
  
  
  RESULTS
  The best condition for this assay are: cell density as 4 x 10(5)/mL, stimulating time as 4 hours, luciferase substrate as 50 microL. 50% effective dose (ED50) of IL-22 assayed by this method is 16.89 ng/mL, relative standard deviation (RSD) is 7.09%. Neutralizing antibody test shows the high specificity. Comparing with ELISA, the method described here has more advantages, including higher stability, easier performance and less cost.
  
  
  CONCLUSIONS
  Luciferase reporter gene assay method is a fast, sensitive, reproducible method for IL-22 bioactivity determination.
authors.affiliation: Department of Biochemistry and Molecular Biology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
authors.forename: Jun-Yao
authors.initials: JY
authors.lastname: Zhang
authors.forename: Zhi-Hua
authors.initials: ZH
authors.lastname: Huang
authors.forename: Yu-Liang
authors.initials: YL
authors.lastname: Huang
authors.forename: Fei
authors.initials: F
authors.lastname: Xu
authors.forename: Xiao-Ying
authors.initials: XY
authors.lastname: Li
authors.forename: Rui-Qing
authors.initials: RQ
authors.lastname: Xing
authors.forename: Qiang
authors.initials: Q
authors.lastname: Fu
mesh_terms: D001681:Biological Assay / Q000379:methods*; D017930:Genes, Reporter; D056945:Hep G2 Cells; D006801:Humans; D007378:Interleukins / Q000032:analysis*; D008156:Luciferases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis*; D050796:STAT3 Transcription Factor / Q000235:genetics*; D000097043:Interleukin-22
chemical_list: D007378:Interleukins; D008156:Luciferases; D011994:Recombinant Proteins; D050796:STAT3 Transcription Factor; D000097043:Interleukin-22