title: [Preparation and characterization of monoclonal antibodies against S100A9].
abstract: OBJECTIVE
  To prepare monoclonal antibodies (mAbs) against S100A9, and characterize these antibodies' properties for functional studies.
  
  
  METHODS
  We amplified cDNA fragment of S100A9 from adult human liver sample, then constructed the vectors pGEX-4T-1-S100A9 and pET-32a-S100A9 for protein expression. The S100A9 protein fused respectively with the His-tag and GST-tag were expressed in E.coli and purified for immunizing mice as antigen and hybridoma screening, respectively. After hybriodma screening, the titer of mAbs was determined by ELISA and the specificity was identified by Western blotting and indirect immunofluorescence technique.
  
  
  RESULTS
  Both GST-tagged and His-tagged S100A9 fusion proteins were successfully constructed and expressed in E.coli. Eighteen hybridoma cell strains secreting S100A9 mAbs were obtained, of which 15 showed strong positive reaction but 3 showed weak positive reaction to the recombinant protein in Western blotting. Two hybridoma cell strains were capable of binding to S100A9 native protein in tumor tissue interstitial fluid.
  
  
  CONCLUSIONS
  We successfully prepared the mAbs against S100A9 protein, which provide the useful tool for further study on biological function and clinical detection of S100A9.
authors.affiliation: Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing 102206, China.
authors.forename: Yangyang
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing 102206, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing 102206, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zuo
authors.affiliation: Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing 102206, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chi
authors.affiliation: Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing 102206, China.
authors.forename: Gang
authors.initials: G
authors.lastname: Liu
authors.affiliation: Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing 102206, China.
authors.forename: Longqin
authors.initials: L
authors.lastname: Sun
authors.affiliation: Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing 102206, China.
authors.forename: Junyi
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Department of Antibody Engineering, Beijing Proteome Research Center, State Key Laboratory of Proteomics, Beijing 102206, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Liu
mesh_terms: D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D040502:Calgranulin B / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007106:Immune Sera / Q000276:immunology*; D008099:Liver / Q000378:metabolism; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D011485:Protein Binding / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D040502:Calgranulin B; D007106:Immune Sera; D011993:Recombinant Fusion Proteins