title: [Purification of human endothelial overexpressed lipopolysaccharide-associated factor 1 protein].
abstract: OBJECTIVE
  To investigate the feasibility of obtaining of a highly pure protein of human endothelial overexpressed lipopolysaccharide-associated factor 1 (EOLA1) with metal chelation chromatography.
  
  
  METHODS
  Inclusion bodies of the E. coli transformed with EOLA1 gene were extracted and washed with BugBuster Protein Extraction Reagent. The primary purified products were purified by His. Bind Resin Chromatography under denaturing condition and dialyzed for renaturation, and then were analyzed with SDS-PAGE, Western blotting and peptide mass fingerprinting (PMF).
  
  
  RESULTS
  EOLA1 was mainly expressed in E. coli as insoluble inclusion bodies. The protein content in the primary extracted inclusion bodies accounted for over 75%, and it accounted for more than 90% after chromatography and renaturation. It was indicated by PMF that the targeted protein peptide overlaid many of designed protein peptide.
  
  
  CONCLUSIONS
  The method of EOLA1 protein purification and renaturation was convenient and efficient, and by this method sufficient amount of highly pure EOLA1 protein could be obtained for the preparation of EOLA1 monoclonal antibody and for the study of its gene function.
authors.affiliation: Institute of Burn Research , Southwest Hospital , State Key Laboratory of Trauma , Burns and Combined Injury, The Third Military Medical University, Chongqing 400038 , P. R. China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Cai
authors.forename: Zi-wen
authors.initials: ZW
authors.lastname: Liang
authors.forename: Xiang-dong
authors.initials: XD
authors.lastname: Luo
authors.forename: Zong-cheng
authors.initials: ZC
authors.lastname: Yang
authors.forename: Rong-ju
authors.initials: RJ
authors.lastname: Sun
authors.forename: Yong-yue
authors.initials: YY
authors.lastname: Su
mesh_terms: D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D042783:Endothelial Cells / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; C495142:EOLA1 protein, human; D008565:Membrane Proteins