ELISA for semicarbazide and its application for screening in food contamination

Analytica Chimica Acta
M VassM Franek

Abstract

The development of a direct competitive enzyme-linked immunosorbent assay (ELISA) based on polyclonal antibodies specific for semicarbazide (SEM) is described. Molecular modelling of the hapten mimics and other key components of the assay system was conducted to explain antibody properties in relation to hapten design. The small aliphatic molecule SEM was coupled to 3-carboxybenzaldehyde to produce carboxyphenyl-SEM (CPSEM), for the generation of specific antibodies. Five rabbits produced antibodies against NPSEM (used in direct and indirect ELISA formats) exhibiting a 50% binding inhibition level (IC(50) values) of 0.06-2.28 microgL(-1) in assay buffer for SEM. The most sensitive indirect assay based on the antibody MVK39 showed a high dynamic range providing a linear readout in the range of 0.01-0.2 microgL(-1). Antibody MVK31 (IgG) allowed specific SEM detection at an IC(50)=0.14 microgL(-1) in direct ELISA and was evaluated using solvent extracted SEM-spiked porcine and baby food samples. Recovery levels determined from fortified samples (0.5, 1.0, 1.5, 5, 10 and 20 microgkg(-1)) of porcine and baby food ranged from 82.9 to 105.3%, respectively, with a coefficient of variation less than 15.5%. Respective detection capabilit...Continue Reading

References

Sep 18, 2004·Journal of Agricultural and Food Chemistry·Adam BecalskiStephen W Seaman
Jun 11, 2005·Analytical and Bioanalytical Chemistry·Maria Beatriz de la Calle, Elke Anklam
Feb 2, 2006·Food Additives and Contaminants·A BecalskiS Seaman
Jul 9, 2008·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·Maria VassMilan Franek

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Citations

Mar 12, 2011·Journal of Agricultural and Food Chemistry·Xiao Dong YanJian Ping Wang
Jul 9, 2008·Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment·Maria VassMilan Franek
Oct 4, 2012·Journal of Environmental Science and Health. Part. B, Pesticides, Food Contaminants, and Agricultural Wastes·Li L WangJian P Wang
Jul 12, 2011·Analytica Chimica Acta·Roberta GalariniFrancesca Lega
Feb 24, 2009·Journal of Agricultural and Food Chemistry·Jun LiJian Ping Wang
Feb 10, 2009·Journal of Agricultural and Food Chemistry·Saowapa ChumaneePakawadee Sutthivaiyakit
Jun 10, 2010·Journal of Agricultural and Food Chemistry·Vanesa JiménezRamon Companyó

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