title: Evaluation of a newly developed quantitative heart-type fatty acid binding protein assay based on fluorescence immunochromatography using specific monoclonal antibodies.
abstract: OBJECTIVE
  To develop a rapid, sensitive and specific assay for quantification of serum heart-type fatty acid binding protein (H-FABP) based on immunofluorescence of specific monoclonal antibodies.
  
  
  METHODS
  We generated novel H-FABP-directed monoclonal antibodies by cloning of spleen cells of mice immunized with H-FABP. Epitopes were mapped and antigen affinity was assessed by surface plasmon resonance (SPR). The H-FABP specific monoclonal antibodies were coupled to fluorescent beads and sprayed onto a nitrocellulose membrane facilitating quantification of H-FABP by immunofluorescence. Reagent cross-reactivity, interference resistance, accuracy and sensitivity were examined. A total of 103 clinical samples were used to compare the sensitivity and specificity of the new assay to a commercially available Randox kit.
  
  
  RESULTS
  This new assay could be finished within 15 min, with sensitivity reaching 1 ng/ml. In a trial of 103 clinical serum samples, the new testing kit results were highly correlated with those from the Randox kit (R(2) = 0.9707). Using the Randox kit as the reference kit, the sensitivity of the new assay was 98.25%, and specificity was 100%.
  
  
  CONCLUSIONS
  An immunofluorescence-based H-FABP assay employing novel monoclonal antibodies could rapidly, specifically and sensitively detect H-FABP in serum samples, providing an effective method for rapid clinical assessment of H-FABP index in the clinic.
authors.affiliation: a School of Bioscience and Bioengineering, South China University of Technology, Guangzhou , Guangdong Province , China.|b National Engineering Laboratory of Point-of-Care Testing, Guangzhou Wondfo Biotech Co. Ltd , Guangzhou, Guangdong Province , China.
authors.forename: Keren
authors.initials: K
authors.lastname: Kang
authors.affiliation: b National Engineering Laboratory of Point-of-Care Testing, Guangzhou Wondfo Biotech Co. Ltd , Guangzhou, Guangdong Province , China.
authors.forename: Peidian
authors.initials: P
authors.lastname: Wu
authors.affiliation: a School of Bioscience and Bioengineering, South China University of Technology, Guangzhou , Guangdong Province , China.|b National Engineering Laboratory of Point-of-Care Testing, Guangzhou Wondfo Biotech Co. Ltd , Guangzhou, Guangdong Province , China.
authors.forename: Wenmei
authors.initials: W
authors.lastname: Li
authors.affiliation: b National Engineering Laboratory of Point-of-Care Testing, Guangzhou Wondfo Biotech Co. Ltd , Guangzhou, Guangdong Province , China.
authors.forename: Shixing
authors.initials: S
authors.lastname: Tang
authors.affiliation: b National Engineering Laboratory of Point-of-Care Testing, Guangzhou Wondfo Biotech Co. Ltd , Guangzhou, Guangdong Province , China.
authors.forename: Jihua
authors.initials: J
authors.lastname: Wang
authors.affiliation: a School of Bioscience and Bioengineering, South China University of Technology, Guangzhou , Guangdong Province , China.
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Luo
authors.affiliation: a School of Bioscience and Bioengineering, South China University of Technology, Guangzhou , Guangdong Province , China.
authors.forename: Mingquan
authors.initials: M
authors.lastname: Xie
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry*; D001667:Binding, Competitive; D015415:Biomarkers / Q000097:blood; D002845:Chromatography; D000074026:Fatty Acid Binding Protein 3; D050556:Fatty Acid-Binding Proteins / Q000097:blood* / Q000276:immunology; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D057230:Limit of Detection; D008807:Mice, Inbred BALB C; D017202:Myocardial Ischemia / Q000097:blood; D011485:Protein Binding
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D015415:Biomarkers; C493408:FABP3 protein, human; D000074026:Fatty Acid Binding Protein 3; D050556:Fatty Acid-Binding Proteins
keywords: H-FABP; Myocardial reperfusion injury; immunochromatography; molecular diagnostics; point-of-care testing