Journal of Clinical and Cellular Immunology
Open Access
ISSN: 2155-9899
ISSN: 2155-9899
Quoc Anh Bui, Minh Tam Duong, Vo Thi Hong and My Linh Bui
Saigon Hi-Tech Park, Vietnam
Posters & Accepted Abstracts: J Clin Cell Immunol
The adulteration of melamine into food is banned but its misuse have been widely reported in milk products and animal feed. The development of a rapid screening immunoassay for monitoring these substances is an urgent requirement. In this research, a hapten of melamine was successfully synthesized by introducing three carbon chain length spacer arm via a reaction between 6-aminocaproic acid and 2-chloro-4,6-diamino-1,3,5-triazine (CAAT). The molecular structure of this hapten was identified by H1 nuclear magnetic resonance spectrometry. An immunogen was prepared by coupling this hapten to Keyhole limpet hemocyanin (KLH). An enzyme conjugated was prepared by coupling synthesized hapten to Horseradish peroxidase (HRP). A direct competitive ELISA (cELISA) was developed to evaluate assay performance. The results showed that high titer polyclonal antibodies were obtained and the performance of this cELISA kit showed good specification: IC50: 230 ng mL-1, LOD: 8.3 ng mL-1 and LOQ: 10 ng mL-1. Regarding selectivity performance, no obvious cross-reactivity to common compound was found. This data show that this ELISA kit met the requirement of melamine maximum residues level (1000 ng mL-1 in milk products, 2500 ng mL in animal feed) and could be use in an immunoassay for the rapid and sensitive detection of this banned chemical.
Email: quocanhchem@gmail.com