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Development of a highly sensitive and specific monoclonal antibody based enzyme‐linked immunosorbent assay for the detection of a new β‐agonist, phenylethanolamine A, in food samples
Author(s) -
Jiang Danni,
Cao Biyun,
Wang Meiyu,
Yang Hong,
Zhao Kang,
Li Jianguo,
Li Mingxin,
Sun Lulu,
Deng Anping
Publication year - 2017
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/jsfa.7826
Subject(s) - monoclonal antibody , chemistry , agonist , chromatography , phenylethanolamine , immunoassay , antibody , enzyme , detection limit , homogeneous , coefficient of variation , microbiology and biotechnology , biochemistry , receptor , biology , immunology , tyrosine hydroxylase , physics , thermodynamics
BACKGROUND All β‐agonists are banned as feed additives for growth promotion in animals due to toxic effects on humans after consuming the β‐agonist contaminated meats. Phenylethanolamine A ( PA ) is a newly emerged β‐agonist. Thus there is a need to develop highly sensitive and specific analytical methods for the detection of PA in food samples. In this study, the monoclonal antibody ( mAb ) against PA was produced by hybridoma technology and used for the development of enzyme‐linked immunosorbent assay ( ELISA ). RESULTS The IC 50 values and limits of detection ( LODs ) of the ELISA using homogeneous combination of coating antigen/antibody for PA were 0.16 ng mL −1 and 0.011 ng mL −1 , respectively. The cross‐reactive ( CR ) values of the assay with 14 structurally related β‐agonists were lower than 0.59%. Swine liver and meat samples were spiked with PA at different content and analysed by ELISA . Acceptable recovery rates of 91.40–105.51% and intra‐assay coefficients of variation of 1.56–9.92% ( n = 3) were obtained. The ELISA for seven spiked samples was confirmed by LC‐MS / MS with a high correlation coefficient of 0.9881. CONCLUSION The proposed mAb ‐based ELISA was highly sensitive and specific for PA and could be used as a quantitative/screening method for PA analysis in food samples. © 2016 Society of Chemical Industry