Journal
ANALYTICAL CHEMISTRY
Volume 88, Issue 7, Pages 3909-3916Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.6b00082
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Funding
- Natural Science Foundation of China [U1301214]
- Guangdong Natural Science Foundation [S2013030013338, 2015A030313366]
- Guangdong Planed Program in Science and Technology [2014TX01N250, 2010A032000001-4, 2013B051000072]
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On the basis of the structural features of (fluoro)quinolones (FQs), pazufloxacin was first used as a generic immunizing hapten to raise a broad-specificity antibody. The obtained polyclonal antibody exhibited :broad cross-reactivity ranging from 5.19% to 478.77% with 21 FQs. Furthermore, the antibody was able to recognize these FQs below their maximum residue limits (MRLs) in an indirect competitive chemiluminescence enzyme immunoassay (ic-CLEIA), with the limit of detection (LOD) ranging from 0.10 to 33.83 ng/mL. For simply pretreated milk samples with spiked FQs, the ic-CLEIA exhibited an excellent recovery with a range of 84.6-106.9% and an acceptable coefficient of variation below 15%, suggesting its suitability and reliability for the use of a promising tool to detect FQs. Meanwhile, comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) models, with statistically significant correlation coefficients (q(CoMFA)(2) = 0.559, r(CoMFA)(2) = 0.999; q2(CoMSIA) = 0.559, r(2CoMSIA) = 0.994), were established to investigate the antibody recognition mechanism. These two models revealed that in the antibody, the active cavity binding FQs' 7-position substituents worked together with another cavity (binding FQs' 1-position groups) to crucially endow the high cross-reactivity. This investigation will 'be significant for better exploring the recognition mechanism and for designing new haptens.
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