- Nifursol hapten, artificial antigen, and preparation method and application thereof
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The invention relates to the field of biochemical engineering, and specifically relates to a nifursol hapten, an artificial antigen, and a preparation method and application thereof. The structure of the nifursol hapten is shown as a formula (I), the artificial antigen is obtained after the nifursol hapten is coupled with carrier protein, the artificial antigen is used as an immunogen to prepare a corresponding hybridoma cell 3H10, and a monoclonal antibody secreted by the hybridoma cell 3H10 can be used for detecting nifursol. A monoclonal antibody prepared by immunizing a mouse with an artificial antigen prepared from the nifursol hapten provided by the invention has relatively high sensitivity in detection of nifursol and derivatives thereof, and a new material basis is provided for establishing a rapid, simple, cheap, sensitive and specific nifursol detection method.
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- Design and synthesis of immunoconjugates and development of an indirect ELISA for rapid detection of 3, 5-dinitrosalicyclic acid hydrazide
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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 acid-ovalbumin (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.
- Shen, Yu-Dong,Zhang, Shi-Wei,Lei, Hong-Tao,Wang, Hong,Xiao, Zhi-Li,Jiang, Yue-Ming,Sun, Yuan-Ming
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p. 2238 - 2248
(2008/12/23)
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