Immunoadsorbent purification of carcinoembryonic antigen using a monoclonal antibody: a direct comparison with a conventional method.
Carcinoembryonic antigen (CEA) has been extracted and purified by two different methods from a single aqueous homogenate of liver metastases from a primary adenocarcinoma of the colon, thus enabling direct comparison to be made between a conventional technique using gel filtration and a monoclonal antibody immunoadsorbent method. Yield, immunoreactivity and purity have been determined in both cases by direct weighing, enzyme-linked immunoadsorbent assay, radioimmunoassay and SDS-polyacrylamide gel electrophoresis. Reactivity of the antigen with monoclonal and polyclonal anti-CEA antibodies recognising different epitopes was analysed by Western blotting. There appears to be no significant difference in immunoreactivity or purity by these criteria, but the immunoadsorbent method gave a higher yield of CEA for far less expenditure of time and effort. A variant of CEA with a lower molecular weight was also identified in both preparations.
Ford
C H
CH
Oncology Research, Memorial University, St. John's, Newfoundland, Canada.
Macdonald
F
F
Griffin
J A
JA
Life
P
P
Bartlett
S E
SE
D000911:Antibodies, Monoclonal / Q000276:immunology; D002272:Carcinoembryonic Antigen / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D007163:Immunosorbent Techniques; D066298:In Vitro Techniques; D008113:Liver Neoplasms / Q000556:secondary
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen