Structural Changes Between Beef Oxy- And Metmyoglobin as Revealed by Monoclonal Antibodies

Oxy- and metmyoglobin were purified from beef muscle using ion-exchange chromatography on Mono-Q. Their reactivity against six monoclonal antibodies (MAbs) to beef myoglobin was compared in a competitive ELISA using biotinylated beef oxymyoglobin. Four MAbs reacted with higher affinity for oxymyoglobin than for metmyoglobin. A similar higher reactivity of oxymyoglobin versus metmyoglobin was obtained with a sandwich ELISA using peroxidase-labelled rabbit anti-serum against beef myoglobin. Such a result has never been reported for myoglobins of any species. These MAbs should be valuable tools for analyzing conformational changes of oxymyoglobin solutions when subjected to chemical or physical treatments.