Development of an Elisa to detect melamine in milk product

Abstract

Research outlines the application of thedirect competitiveenzyme-linked immunosorbent assay (ELISA) for the analysis of melamine in milk products. Compared with other instrumental methods such as liquid chromatography-mass spectrometry (LC-MS), gas chromatography-mass spectrometry (GC-MS), ELISA has many advantages: sensitivity, simplicity and cost-effectiveness. The results of the study show that, two haptens were synthesized.It was found that the ELISA show best sensitivity and specificity to melamine when using the coating antigen prepared by coupling 2-(aminocaproic acid)-4,6-diamino-1,3,5–triazine with ovalbumin (OVA) .The antibodies were used to produce ELISA test kits to detect melamine residues. The 50% inhibitory concentration (IC50) was 139 µgxkg−1. The limit of detection (LOD) was 3 µgxkg−1. The detectable working range (20–80% inhibitory concentration) was from 7.62-µgxkg−1 to 2519µgxkg−1, respectively. Keywords: ELISA, Melamine, Test kits,Milk, hapten.