Characterization of a monoclonal antibody against a mucin-type glycoprotein in human sweat.
We report the preparation and characterization of an IgG2 monoclonal antibody (MAb), HSMA, prepared against a human pooled sweat extract (HPSE). The major component of HPSE was a mucin-type molecule, as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) stained with periodic acid-Schiff (PAS) reagent. By immunoblotting, HSMA revealed a smear in the high molecular weight range, typical of mucins. In enzyme-linked immunosorbent assay (ELISA), HSMA failed to react with HPSE fractions isolated after anionic exchange gel chromatography. Similarly, radio-immunobinding assays demonstrated no reactivity between HSMA and A, B, H, and Lewis blood group-related structures. The immunohistological labeling on normal skin showed that HSMA reacted with the cells of eccrine sweat glands, and to a lesser extent, with sebaceous glands and epidermal cells. Periodate treatment in situ abolished these reactions, thus suggesting the carbohydrate structure of the HSMA-epitope. In indirect immunofluorescence (IF) studies, HSMA also reacted with other exocrine glands, e.g. mammary glands, sublingual glands, mixed sero-mucous glands of the trachea, and in the pancreas. Sparse positive cells were also observed in the testis, kidney, thyroid and digestive tract.
Dupuy
P
P
Department of Dermatology, Royal Victoria Hospital, McGill University, Montreal, Quebec, Canada.
LePendu
J
J
Jothy
S
S
Wilkinson
R D
RD
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000941:Antigens; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D009077:Mucins / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D012867:Skin / Q000276:immunology / Q000378:metabolism; D013542:Sweat / Q000737:chemistry / Q000276:immunology*; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D000941:Antigens; D009077:Mucins