Broad-Specificity Chemiluminescence Enzyme Immunoassay for (Fluoro)quinolones: Hapten Design and Molecular Modeling Study of Antibody Recognition

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• 作者：Haopeng Zeng ; Jiahong Chen ; Chijian Zhang ; Xin-an Huang ; Yuanming Sun ; Zhenlin Xu ; Hongtao Lei
• 刊名：Analytical Chemistry
• 出版年：2016
• 出版时间：April 5, 2016
• 年：2016
• 卷：88
• 期：7
• 页码：3909-3916
• 全文大小：429K
• ISSN：1520-6882

文摘

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 (q²_CoMFA = 0.559, r²_CoMFA = 0.999; q²_CoMSIA = 0.559, r²_CoMSIA = 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.