Analytical Biochemistry 2012-04-15

Production of the monoclonal antibody against Sudan 2 for immunoassay of Sudan dyes in egg.

Jing Liu, Huicai Zhang, Dongsheng Zhang, Feng Gao, Jianping Wang

Index: Anal. Biochem. 423(2) , 246-52, (2012)

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Abstract

Many methods have been reported to determine the residues of Sudan dyes in food samples. Among the reported methods, enzyme-linked immunosorbent assay (ELISA) was a frequently used practical screen tool. In this study, a novel hapten of Sudan 2 was synthesized by coupling 4-amino-3-methylbenzoic acid to β-naphthol, and the monoclonal antibody against Sudan 2 was produced. The obtained antibody can recognize Sudan 1, 2, 3, and 4, Sudan red G, and Para red simultaneously. After evaluation of different coating antigens, a heterologous indirect competitive ELISA was then developed to determine the six Sudan dyes in egg. The cross-reactivities for the six analytes were in a range of 63% to 100%, and the limits of detection were in a range of 0.2 to 0.5 ng/g, depending on the compound. Intra- and interassay recoveries from the standard fortified blank eggs were in a range of 71.7% to 97.6% with coefficients of variation lower than 17.1%.Copyright © 2012 Elsevier Inc. All rights reserved.