Development of Monoclonal Antibodies and Indirect Competitive Enzyme-Linked Immunosorbent Assay Kits for the Detection of Clenbuterol and Salbutamol in the Tissues and Products of Food-Producing Animals

Dapeng Peng, Lingyan Zhang, Chen Situ, Yuanhu Pan, Yanfei Tao, Yulian Wang, Zonghui Yuan

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6 Citations (Scopus)

Abstract

To better monitor the residues of clenbuterol and salbutamol in edible tissues and products of animals treated with these compounds, a monoclonal antibody (mAb) against the β-agonists was prepared, and an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on this antibody was also developed. The hapten of salbutamol was synthesized and conjugated to carrier proteins via different linkers. Female Balb/c mice were immunized with the hapten-protein conjugates to produce monoclonal antibodies using the standard fusion procedures. After fusion and four cloning cycles, eight hybridomas were isolated and of which one clone, 6E5, that has the isotype IgG1, was selected for the present study. The cross-reactivities of the mAb 6E5 towards salbutamol, clenbuterol, mabuterol, cimaterol, clenproperol, mapenterol, tuoloterol and terbutaline were determined as 104.5, 100, 100, 83.6, 71.9, 44.2, 6.3 and 2.3%, respectively. The limit of detection for salbutamol and clenbuterol in edible animal tissues was 0.21 and 0.57 μg kg−1, respectively. The recoveries ranged from 55.4 to 122.0%, and the CVs were less than 19.6%. When used with spiked or incurred samples, or in a field trial, the established ic-ELISA has demonstrated a consistent performance in various biological matrices, suggesting this is a sensitive, accurate and low-cost analytical method.
Original languageEnglish
JournalFood Analytical Methods
Early online date18 May 2017
DOIs
Publication statusEarly online date - 18 May 2017

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