Demonstration of two molecular variants of carcinoembryonic antigen by concanavalin A sepharose affinity chromatography.
The carcinoembryonic antigen (CEA) active glycoproteins from perchloric acid extract of liver-metastasized primary colon tumor have been separated by concanavalin A Sepharose (Con A Sepharose) chromatography. The CEA activities separated by Con A Sepharose chromatography were designated as loosely bound and tightly bound which, respectively, eluted on the Con A Sepharose column between 0.12 and 0.15 M and 0.3 M alpha-methylmannose in a linear gradient of alpha-methylmannose. Further purification of these activities by Sephadex G-200, Bio-Gels A-1.5m and P-300 yielded two variants of glycoproteins (B1 and C2) with CEA activity. Both purified preparations of CEA had similar immunochemical properties. Their A280/A260 ratios were 1.30 and 1.56, respectively. The purified loosely bound CEA (B1) had immunological, chromatographic, and electrophoretic properties similar to those of 125I-CEA, whereas the tightly bound CEA (C2) had a lower molecular weight (120,000 to 140,000). Further, specificity to these two CEA's was established by their reactions in immunoelectrophoresis with preparations of specific goat anti-CEA anti-serum obtained from other investigators. The results indicate the practical use of Con A Sepharose affinity chromatography for the separation and characterization of glycoprotein tumor antigens.
D002272:Carcinoembryonic Antigen / Q000032:analysis* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003110:Colonic Neoplasms / Q000276:immunology; D003208:Concanavalin A; D000939:Epitopes; D006023:Glycoproteins / Q000032:analysis / Q000302:isolation & purification; D006801:Humans; D009362:Neoplasm Metastasis; D011863:Radioimmunoassay
D002272:Carcinoembryonic Antigen; D000939:Epitopes; D006023:Glycoproteins; D003208:Concanavalin A