Abstract
Twenty-five phages that selectively bind to a monoclonal antibody (Mab) 1H2 specific to 2,2′,4,4′-tetrabromodiphenyl ether (BDE47) in the absence or presence of BDE47 have been selected from phage‐display libraries containing cyclic 7-mer, linear 7-mer, and linear 12-mer randomized peptides. Competitive and noncompetitive enzyme-linked immunosorbent assays (ELISA) for BDE47 were developed by using a clone C7-1 specific to the BDE47-free Mab 1H2 and a clone XC7-8 specific to the BDE47-bound Mab 1H2, respectively. The half-maximum signal inhibition concentration (IC50) of the competitive phage ELISA and the half-maximum signal enhancement concentration (EC50) of the noncompetitive phage ELISA for BDE47 were 6.8 ng mL−1 and 4.2 ng mL−1, respectively. The noncompetitive phage ELISA showed higher cross-reactivity with BDE28, BDE99, and BDE100 than the competitive one, ranging between 1.3 and 6.5 % versus 0.3 and 0.8 %. Recoveries of the competitive and the noncompetitive phage ELISAs for BDE47 in sewage sludge and fillet samples were 96–124 % and 97–120 %, respectively. The results of the two types of phage ELISAs for BDE47 in the real-world samples agreed well with a gas chromatography/electron capture detector-ion trap mass spectrometer method.
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Acknowledgments
This work was supported in part by the National Natural and Science Foundation (NNSF) of China (20977111), the Chinese University Scientific Fund and the Special Project on the Integration of Industry, Education and Research of Guangdong Province (2012B091100281).
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Wang, J., Liu, Z., Li, G. et al. Simultaneous development of both competitive and noncompetitive immunoassays for 2,2′,4,4′-tetrabromodiphenyl ether using phage-displayed peptides. Anal Bioanal Chem 405, 9579–9583 (2013). https://doi.org/10.1007/s00216-013-7364-5
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DOI: https://doi.org/10.1007/s00216-013-7364-5