Antibody to rabbit macrophage type V collagenase/gelatinase and its use to further characterize the enzyme.

A highly specific antiserum was raised against purified rabbit alveolar macrophage type V collagenase/gelatinase. The reactivity of the antiserum was tested in ELISA and was detected at a high titer. Competitive inhibition ELISA demonstrated a linear inhibition of reactivity by pure antigen if either pure antigen or crude culture medium was used as the solid phase antigen. In the latter case, greater than 90% inhibition was achieved indicating a high degree of specificity of the antibody. Immobilized antibody was successful in absorbing the type V collagenolytic activity. Immunoblot analysis of crude culture medium revealed a single immunoreactive band which correlated with both the stained gel of the purified antigen as well as the only zone of gelatinolysis in a gelatin/acrylamide gel. This monospecific antibody will be a useful tool in the further characterization of this enzyme as well as the delineation of its relative role in chronic inflammation.