Journal of Food, Agriculture and Environment
Establishment of a sensitive indirect competitive ELISA standard curve based on the artificial antigen of clofencet
Author(s):
Xin-liang Zhao 1, 2,
Long-yu Zhang 1, Zhi-kuan Li 1, Zheng Chen 1, Gai-sheng Zhang 1*, Qi-di Zhu 1Recieved Date: 2013-05-02, Accepted Date: 2013-09-26
Abstract:To establish an indirect competitive ELISA (icELISA) method for detecting clofencet (SQ-1) residue in wheat samples, NHS ester technology was employed to synthesize the artificial antigen and Balb/C female mice were immunized afterwards to prepare anti-SQ polyclonal antibody. Under optimal conditions, the established icELISA exhibited a working range from 0.083 to 65.2 ng/mL, with the half maximal inhibitory concentration (IC50) and limit of detection (LOD) values of 0.32 ng/mL and 0.05 ng/mL, respectively. After optimization, 1:20 dilution in wheat samples gave an inhibition curve almost the same as that in PBS buffer, and the correlation coefficient (R2) between icELISA and HPLC was 0.9947. The results suggest this icELISA provides a sensitive and cost-effective screening method for detecting SQ-1 residue in wheat samples, and demonstrates a noticeable advantage over methods requiring a tedious sample cleanup procedure.
Keywords:Clofencet,
artificial antigen, polyclonal antibody, indirect competitive ELISA, wheatJournal: Journal of Food, Agriculture and Environment
Year: 2013
Volume: 11
Issue: 3&4
Category: Agriculture
Pages: 1093-1096
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