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Molecules 2008, 13(9), 2238-2248; doi:10.3390/molecules13092238

Design and Synthesis of Immunoconjugates and Development of an Indirect ELISA for Rapid Detection of 3, 5-Dinitrosalicyclic Acid Hydrazide

2,*  and 2,*
1 College of Food Science, South China Agricultural University, Guangzhou 510642, P. R. China 2 South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, P. R. China
* Authors to whom correspondence should be addressed.
Received: 22 August 2008 / Revised: 14 September 2008 / Accepted: 16 September 2008 / Published: 23 September 2008
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In this study novel immunoconjugates were designed, synthesized and then used to develop a rapid, specific and sensitive indirect ELISA method to directly detect residues of 3,5-dinitrosalicyclic acid hydrazide (DNSH), a toxic metabolite of nifursol present in chicken tissues. The hapten DNSHA was first designed and used to covalently couple to BSA to form an immunogen which was immunized to rabbits to produce a polyclonal antibody against DNSH. Furthermore, a novel 3,5-dinitrosalicylic acidovalbumin (DNSA-OVA) immunoconjugate structurally different from DNSHA-OVA was designed and used as a “substructural coating antigen” to improve the sensitivity of an indirect ELISA analysis for a direct DNSH detection. Based on the “substructural coating antigen” concept, an optimized indirect ELISA method was established that exhibited good specificity and high sensitivity for detecting DNSH, with a cross-reactivity of less than 0.1% (excluding the parent compound nifursol), IC50 of 0.217 nmol/mL and detection limit of 0.018 nmol/mL. Finally, a simple and efficient analysis of DNSH samples in chicken tissues showed that the average recovery rate of the indirect ELISA analysis was 82.3%, with the average coefficient of variation 15.9%. Thus, the developed indirect ELISA method exhibited the potential for a rapid detection of DNSH residues in tissue.
Keywords: Nifursol; DNSH; Hapten design; Indirect ELISA Nifursol; DNSH; Hapten design; Indirect ELISA
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Shen, Y.-D.; Zhang, S.-W.; Lei, H.-T.; Wang, H.; Xiao, Z.-L.; Jiang, Y.-M.; Sun, Y.-M. Design and Synthesis of Immunoconjugates and Development of an Indirect ELISA for Rapid Detection of 3, 5-Dinitrosalicyclic Acid Hydrazide. Molecules 2008, 13, 2238-2248.

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