Characterization of several monoclonal antibodies generated against ferret tracheal epithelial cells.
Monoclonal antibodies have been generated to ferret tracheal epithelial (FTE) cells. Four of the antibodies exhibiting positive reactivity in a fluorescence-activated cell sorter were further characterized. In tracheal sections, three of the antibodies reacted with submucous gland cells and material intermeshed in the cilia while the fourth antibody failed to react with epithelial cells. The three antibodies which reacted in tracheal sections also reacted with void volume fractions following Sepharose CL-6B chromatography of a solubilized FTE cell preparation. Reactivity of the void volume fractions to these antibodies was sensitive to periodate oxidation, and unaffected by heating (100 degrees C X 30 min) or methanol extraction suggesting a carbohydrate epitope. The fourth antibody recognized periodate-insensitive antigen in the void volume and included column fractions. Binding of one of the antibodies to periodate-sensitive antigen was completely blocked by 10 mM N-acetyl galactosamine as demonstrated in an ELISA with fixed FTE cells. In a western blot analysis, two of the antibodies which recognized periodate-sensitive antigen also reacted with a large heterogenous macromolecule.
Groelke
J W
JW
Coalson
J J
JJ
Baseman
J B
JB
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000941:Antigens / Q000032:analysis; D004847:Epithelial Cells; D005260:Female; D005289:Ferrets; D005434:Flow Cytometry; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D009092:Mucous Membrane / Q000166:cytology; D014132:Trachea / Q000166:cytology*
D000911:Antibodies, Monoclonal; D000941:Antigens; D007074:Immunoglobulin G