STRUCTURE OF NATIVE AND HEAT-DENATURED OVALBUMIN AS REVEALED BY MONOCLONAL-ANTIBODIES - EPITOPIC CHANGES DURING HEAT-TREATMENT

Twenty-one monoclonal antibodies (MAbs) of IgG1 or IgM isotype were developed against either native or heat denatured ovalbumin. Most of the anti-native ovalbumin MAbs were of the IgG1 isotype and had high affinity for native ovalbumin in a sandwich ELISA test (antigen bound by immunocapture with polyclonal antibodies). Anti-native MAbs did not recognize plastic bound ovalbumin in an indirect ELISA. Most anti-heat denatured ovalbumin MAbs were of the IgM isotype and reacted with ovalbumin on plastic but not with ovalbumin in the sandwich ELISA test. An immunocapture test using these MAbs was developed which enabled determination of temperatures to which ovalbumin had previously been heated.