Huiming Sheng1, Zhi-Gang Hu2,3, Jie Liu2, Fenghong Yuan2, Mei Li2, Yaohong Zou2, Yu Chen3. 1. Shanghai Tongren Hospital Affiliated to Shanghai Jiaotong University, School of Medicine, Shanghai, China. 2. Affiliated Wuxi People's Hospital of Nanjing Medical University, Wuxi, China. 3. Department of Laboratory Medicine, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
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%. CONCLUSION: 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.
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%. CONCLUSION: 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: Farah T van Genderen; Frans K Gorus; Daniel G Pipeleers; Christiaan F H van Schravendijk Journal: Endocrinology Date: 2013-03-24 Impact factor: 4.736
Authors: Robert B M Landewé; Maarten Boers; Arco C Verhoeven; Rene Westhovens; Mart A F J van de Laar; Harry M Markusse; J Christiaan van Denderen; Marie Louise Westedt; Andre J Peeters; Ben A C Dijkmans; Piet Jacobs; Annelies Boonen; Désirée M F M van der Heijde; Sjef van der Linden Journal: Arthritis Rheum Date: 2002-02
Authors: F A van Gaalen; S P Linn-Rasker; W J van Venrooij; B A de Jong; F C Breedveld; C L Verweij; R E M Toes; T W J Huizinga Journal: Arthritis Rheum Date: 2004-03