Cancer-associated glycoproteins defined by a monoclonal antibody, MLS 128, recognizing the Tn antigen.
A murine monoclonal antibody, MLS 128, recognizing the Tn antigen, was established and used for characterization of glycoproteins expressing the Tn antigen. The Tn antigen was expressed on three polypeptide chains with molecular weights of 250k, 210k and 150k daltons. LS 180 cells were labeled with 3H-glucosamine or 35S-sulfate metabolically, and then the immunoprecipitate derived from the cell lysate was subjected to SDS-PAGE followed by fluorography. It was revealed that these Tn antigen glycoproteins were produced through the processing of a high molecular weight precursor. The carbohydrate moieties of the Tn antigen glycoproteins labeled with 3H-glucosamine were released with alkaline-borohydride, and the released sugars were examined by gel filtration and paper chromatography. The carbohydrates predominantly consisted of GalNAc and sialyl GalNAc (greater than 90%), with a nearly equal distribution.
Nakada
H
H
Department of Biotechnology, Faculty of Engineering, Kyoto Sangyo University, Japan.
Inoue
M
M
Numata
Y
Y
Tanaka
N
N
Funakoshi
I
I
Fukui
S
S
Yamashina
I
I
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000276:immunology*; D015295:Antigens, Tumor-Associated, Carbohydrate; D015153:Blotting, Western; D002236:Carbohydrate Conformation; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D005944:Glucosamine; D006023:Glycoproteins / Q000276:immunology* / Q000302:isolation & purification; D007163:Immunosorbent Techniques; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000276:immunology*; D009844:Oligosaccharides / Q000302:isolation & purification; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D015295:Antigens, Tumor-Associated, Carbohydrate; D006023:Glycoproteins; D009363:Neoplasm Proteins; D009844:Oligosaccharides; C040286:Tn antigen; D005944:Glucosamine