title: [Preparation, epitope analysis and clinical application of monoclonal antibodies against heart-type fatty acid binding protein].
abstract: OBJECTIVE
  To prepare and characterize the monoclonal antibody (mAb) against recombinant human heart-type fatty acid binding protein (H-FABP) and apply it to the clinical analysis.
  
  
  METHODS
  BALB/c mice were immunized with recombinant H-FABP (rH-FABP) from prokaryotic expression or synthesized peptide fragment. After common fusion and screening, the subtypes, titer and affinity of mAbs were detected respectively. After purification, the specificity of mAbs was tested by indirect ELISA and Western blotting. ELISA system was established by pair mapping and applied in the detection of clinical samples. Two epitope peptides were designed by bioinformatics and used to detect the epitope of 1-F10 through Western blotting.
  
  
  RESULTS
  Four different hybridoma clones secreting anti-H-FABP antibodies were developed, with high titres of 1:51 200-1:1024 000. Immunoglobulin types of these mAbs were found to be IgG2a or IgG2b, respectively. The affinity of the mAb 1-F10 even reached 9.02×10(9); mol/L. ELISA and Western blotting showed that these mAbs could identify H-FABP specifically. In addition, the pair mapping of monoclonal antibodies 3-H5-1-F10 could recognize H-FABP in human serum samples. Furthermore, the specific target recognized by 1-F10 mAb was located within amino acid 86-133 of H-FABP.
  
  
  CONCLUSIONS
  Four highly specific mAbs against H-FABP were successfully obtained and the established ELISA system could be used to detect the H-FABP in clinical serum samples, and the epitope of 1-F10 mAb was also verified.
authors.affiliation: Department of Clinical Biochemistry, College of Medical Laboratory, Third Military Medical University, Chongqing 400038, China. hanqi688@163.com
authors.forename: Qi
authors.initials: Q
authors.lastname: Han
authors.forename: Shan
authors.initials: S
authors.lastname: Cong
authors.forename: Weijing
authors.initials: W
authors.lastname: Yi
authors.forename: Sha
authors.initials: S
authors.lastname: Chen
authors.forename: Wenbin
authors.initials: W
authors.lastname: Liang
authors.forename: An
authors.initials: A
authors.lastname: Chen
authors.forename: Chuanmin
authors.initials: C
authors.lastname: Hu
authors.forename: Shuhui
authors.initials: S
authors.lastname: Li
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D000917:Antibody Formation; D000918:Antibody Specificity / Q000276:immunology; D003429:Cross Reactions / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology*; D050556:Fatty Acid-Binding Proteins / Q000276:immunology*; D005260:Female; D006801:Humans; D006825:Hybridomas; D051379:Mice; D010446:Peptide Fragments / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D050556:Fatty Acid-Binding Proteins; D010446:Peptide Fragments