Simultaneous development of both competitive and noncompetitive immunoassays for 2,2′,4,4′-tetrabromodiphenyl ether using phage-displayed peptides

Wang, Jia; Liu, Zhiping; Li, Ganqiong; Li, Ji; Kim, Hee-Joo; Shelver, Weilin L.; Li, Qing X.; Xu, Ting
November 2013
Analytical & Bioanalytical Chemistry;Nov2013, Vol. 405 Issue 27, p9579
Academic Journal
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 (IC 50) of the competitive phage ELISA and the half-maximum signal enhancement concentration (EC 50) 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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