Ultrasensitive anti-melamine monoclonal antibody and its use in the development of an immunochromatographic strip

Abstract

The hapten 1 which was formed by the reaction of 2-chloro-4,6-diamine-1,3,5-triazine (CAAT) with 6-aminocaproic acid, conjugated with keyhole limpet haemocyanin to prepare the immunogen. And coating antigen was prepared by the coupling of ovalbumin and hapten 2 which was synthesized by the reaction of CAAT and 3-mercapto propionic acid. Based on immunological and cell fusion technology, the anti-melamine monoclonal antibody was prepared and characterized. The half inhibitory concentration of the antibody for melamine was 1.8 ng/mL, and the linear range was 0.61–5.56 ng/mL. An immunochromatographic (ICT) strip, based on the antibody, was developed for the rapid detection of melamine residues in milk samples. The ICT strip had a visual cut-off value of 10 ng/mL for milk samples. Therefore, the colloidal gold immunoassay with the advantage of high affinity and sensitivity can be used for the ultrasensitive detection of melamine residues in milk samples.

Keywords

• antibody
• immunochromatographic test strip
• melamine
• milk