title: [Purification and separation of mannan-binding lectin (MBL) and MBL-associated serine proteases complex from human plasma].
abstract: OBJECTIVE
  To separate mannan-binding lectin (MBL) and MBL-associated serine proteases (MASPs) from human plasma.
  
  
  METHODS
  A two-step affinity chromatography on underivatized sepharose 4B was employed for purification of MBL-MASP complex, followed by gel filtration on a Sephacryl S-300 column for separation of MBL and MASPs from the complex. The purification procedures were performed at 4 degrees Celsius with the addition of two proteolytic inhibitors, phenyl methylsulfonyl fluoride and 1,10-phenanthroline during affinity chromatography but not in the gel filtration buffer.
  
  
  RESULTS
  Preparations of highly purified MBL and proenzyme MASPs were obtained. The purified MBL was shown by SDS-PAGE and Western blotting to be a functional multimer composed of 28,000 and 32,000 peptide chains, with high bioactivity as demonstrated by ligand-binding assay and yeast agglutination experiment.
  
  
  CONCLUSIONS
  A simple and convenient procedure is established successfully for the purification of MBL and the proenzyme MASPs.
authors.affiliation: Department of Immunology, Southern Medical University, Guangzhou 510515, China. chenyue@fimmu.com
authors.forename: Yue
authors.initials: Y
authors.lastname: Chen
authors.forename: Li-yun
authors.initials: LY
authors.lastname: Zhang
authors.forename: Zheng-liang
authors.initials: ZL
authors.lastname: Chen
mesh_terms: D002846:Chromatography, Affinity; D006801:Humans; D037601:Mannose-Binding Lectin / Q000302:isolation & purification*; D050606:Mannose-Binding Protein-Associated Serine Proteases / Q000302:isolation & purification*; D010949:Plasma / Q000737:chemistry*
chemical_list: D037601:Mannose-Binding Lectin; D050606:Mannose-Binding Protein-Associated Serine Proteases