Development of a Competitive Indirect Enzyme-Linked Immunosorbent Assay Based on Monoclonal Antibodies for the Detection of 2-Dodecylcyclobutanone in Irradiated Beef

文献类型： 外文期刊

第一作者： Zhao, Yueliang

作者： Zhao, Yueliang;Wang, Feng;Li, Weiming;Ha, Yiming

作者机构：

关键词： 2-dodecylcyclobutanone;irradiated beef monoclonal antibody;ELISA

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 影响因子：5.279； 五年影响因子：5.269 ）

ISSN： 0021-8561

年卷期： 2013 年 61 卷 32 期

页码：

收录情况： SCI

摘要： A highly sensitive and specific competitive indirect enzyme-linked immunosorbent assay (ciELISA) based on monoclonal antibodies was developed for the detection of 2-dodecylcyclobutanone (2-DCB), a chemical marker for irradiated lipid-containing foods. 2-0xocyclobutane undecanoic acid was used as an alternative to 2-DCB and conjugated to BSA and OVA via a conventional carbodiimide condensation reaction to prepare the immunogen and the coating antigen for 2-DCB. The monoclonal antibody against 2-DCB was obtained using the hybridoma technique, with a high specificity and low cross-reactivity for 2-tetradecylcyclobutanone (2-TCB; <8%) and other structurally related compounds (<0.1%). The ciELISA method was applicable at optimal experimental conditions of 0.001-100 mu g/mL 2-DCB in a buffer solution, with an IC50 value of 0.25 mu g/rnL and a limit of detection (defined as the IC20) of approximately 0.004 mu g/mL. The recovery efficiency of 2-DCB from ground beef patties ranged from 84.4 to 109.8%. The intra-assay and interassay coefficients of variation were <10.0 and <12.0%, respectively. The proposed method was validated by gas chromatography mass spectrometry with high correlation. The same method was used to detect 2-DCB in ground beef patties irradiated at 0.5, 1.0, 3.0, 5.0, and 7.0 kGy; the 2-DCB concentration linearly increased with the radiation dose.

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