Production and characterization of monoclonal antibodies against human airway mucins

The objective of this study was to generate and characterize monoclonal ant ibodies (MAbs) against human airway mucins, and therefore, should serve as a useful tool in studying the regulation of airway mucins in various physio logical or pathological situations of human airway, As an antigen, we used a high molecular mass mucin preparation purified from the sputum of normal human subjects. Two monoclonal hybridomas, namely MAbs HM02 and HM03 were o btained and they showed strong immunoreactivity against purified or crude m ucin in sputum or bronchial washing off normal human subject. With the high immunoreactivity of these MAbs, mucin contents could be analyzed with more than 100-fold dilution of human airway secretion, The antibodies recognize d carbohydrate epitopes because their immunoreactivity was completely aboli shed by treatment of the mucin with 5 mM periodate, Further characterizatio n of MAbs HM02 and HM03 showed that: (1) they belong to the IgM type; (2) t hey bind to high molecular mass mucins based on Western blot; (3) they coul d indirectly immunoprecipitate human airway mucin and as we know, this is t he first to demonstrate immunoprecipitation of human airway mucin with anti -human mucin antibodies; and (4) they bind to the goblet cell in airway epi thelium as well as some submucosal glands based on immunohistochemistry. Th erefore, MAbs HM02 and HM03 should be able to serve as an invaluable tool i n studying the regulation of airway mucins in various physiological and pat hological situations of human airway.