title: Determination of Anticyclic Citrullinated Peptide Based on Biotin-Streptavidin-Amplified Time-Resolved Fluoroimmunoassay.
abstract: BACKGROUND
  A rapid and sensitive time-resolved fluoroimmunoassay (TRFIA) based on the biotin-streptavidin amplification system was developed for the determination of anticyclic citrullinated peptide (anti-CCP).
  
  
  METHODS
  Europium-labeled streptavidin derivatives combined with europium and anhydride of diethylene triamine pentaacetic acid were used to label streptavidin, biotin was coupled with rabbit anti-human IgG to form a biotin-anti-human IgG bridge between streptavidin-europium and the anti-CCP antibody in the immunoassay. The anti-CCP assay was carried out by measuring the fluorescence of Eu(3+) -streptavidin at 615 nm.
  
  
  RESULTS
  The presented method produced a wide linear range from 0.58 to 9,463 U/ml, while it was only 591.4-18.48 U/ml when using an ELISA kit, and featured a detection limit up to 0.5 U/ml for anti-CCP. The values determined by the biotin-streptavidin-TRFIA and ELISA correlated well (R(2) = 0.8927). The method was applied to determine anti-CCP in serum samples with satisfied recoveries of 96.45-104.63%.
  
  
  CONCLUSIONS
  The assay results obtained by the present method showed that biotin-streptavidin-amplified TRFIA improve the traditional ELISA kit for anti-CCP detection. Therefore, it offers a better alternative immunoassay in rheumatoid arthritis management.
authors.affiliation: Shanghai Tongren Hospital Affiliated to Shanghai Jiaotong University, School of Medicine, Shanghai, China.
authors.forename: Huiming
authors.initials: H
authors.lastname: Sheng
authors.affiliation: Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, China.|Department of Laboratory Medicine, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Hu
authors.affiliation: Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Liu
authors.affiliation: Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, China.
authors.forename: Fenghong
authors.initials: F
authors.lastname: Yuan
authors.affiliation: Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, China.
authors.forename: Mei
authors.initials: M
authors.lastname: Li
authors.affiliation: Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, China.
authors.forename: Yaohong
authors.initials: Y
authors.lastname: Zou
authors.affiliation: Department of Laboratory Medicine, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Chen
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000097:blood*; D001323:Autoantibodies / Q000097:blood*; D001710:Biotin; D016022:Case-Control Studies; D004797:Enzyme-Linked Immunosorbent Assay; D015200:Fluoroimmunoassay; D006801:Humans; D007118:Immunoassay; D057230:Limit of Detection; D008180:Lupus Erythematosus, Systemic / Q000097:blood / Q000175:diagnosis; D008947:Mixed Connective Tissue Disease / Q000097:blood / Q000175:diagnosis; D009103:Multiple Sclerosis / Q000097:blood / Q000175:diagnosis; D010456:Peptides, Cyclic / Q000097:blood* / Q000276:immunology; D011379:Prognosis; D011817:Rabbits; D012595:Scleroderma, Systemic / Q000097:blood / Q000175:diagnosis; D012859:Sjogren's Syndrome / Q000097:blood / Q000175:diagnosis; D019809:Streptavidin / Q000032:analysis*
chemical_list: D000888:Antibodies, Anti-Idiotypic; D001323:Autoantibodies; D010456:Peptides, Cyclic; C048844:anti-IgG; C487763:cyclic citrullinated peptide; D001710:Biotin; D019809:Streptavidin
keywords: cyclic citrullinated peptide; rheumatoid arthritis; time-resolved fluoroimmunoassay