Establishment of a sandwich ELISA for the determination of beef content in processed foods by using monoclonal antibodies to myoglobin

A sandwich enzyme-linked immunosorbent assay specific to beef myoglobin was developed using monoclonal antibodies to sodium dodecyl sulphate-denatured beef myoglobin and a peptide with an amino acid sequence unique to beef myoglobin. This method was very specific for beef myoglobin, and showed no cross-reactivity with not only pork and chicken myoglobins, but also with other food proteins such as egg, cow's milk, wheat and peanut. The limit of detection and that of quantification for the beef protein was 9.5 and 12.8 ng/ml, respectively. When the content of beef protein in pork-and chicken-based model processed foods containing varying amounts of beef were determined by this method, the recovery of beef protein agreed well with the actual ratio of added beef regardless of the processing conditions. Also, the amount of beef protein in several commercially processed foods could be reasonably determined and confirmed by this method.