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 b...

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Bibliographic Details
Published in:Cancer research (Chicago, Ill.) Ill.), 1975-11, Vol.35 (11 Pt 1), p.3001-3008
Main Authors: Harvey, S R, Chu, T M
Format: Article
Language:English
Subjects:
Carcinoembryonic Antigen - analysis
Carcinoembryonic Antigen - isolation & purification
Chromatography, Affinity
Chromatography, Gel
Colonic Neoplasms - immunology
Concanavalin A
Epitopes
Glycoproteins - analysis
Glycoproteins - isolation & purification
Humans
Neoplasm Metastasis
Radioimmunoassay
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Summary: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.
ISSN:0008-5472
1538-7445