Gel filtration of carcinoembryonic antigen (CEA) and non-specific crossreacting antigen (NCA) on the Superose 12HR column: effect of guanidine hydrochloride.
CEA and NCA, which behave as dimeric molecules in SDS-polyacrylamide gel electrophoresis, were submitted to gel filtration on the Superose 12HR column in phosphate-buffered saline (PBS) pH 7.0 or in 6 M guanidine hydrochloride. The elution volumes of these antigens were compared with those of protein markers. The reference proteins showed almost linear relation between log Mr and elution volume, but they were eluted distinctly faster in guanidine that in PBS. The elution volumes of NCA corresponded to Mr approx. 110,000 in PBS and 50,000 in guanidine. These values were accordant with those obtained by other methods and suggested that guanidine dissociates NCA into subunits. The apparent molecular weights of CEA (migrating as 180 kDa molecule in SDS-electrophoresis) estimated by gel filtration on Superose were approx. 700,000 in PBS and 160,000 in guanidine. These results also suggested the dissociation of CEA by guanidine, but due to anomalous elution of CEA from the Superose column they are less convincing than in the case of NCA.
Sedlaczek
P
P
Department of Pathological Anatomy, School of Medicine, Wrocław.
Krop-Watorek
A
A
Gawlikowski
W
W
Lisowska
E
E
D000941:Antigens / Q000302:isolation & purification*; D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen / Q000302:isolation & purification*; D015815:Cell Adhesion Molecules; D002850:Chromatography, Gel; D006023:Glycoproteins / Q000302:isolation & purification*; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D011487:Protein Conformation / Q000187:drug effects; D012685:Sepharose
D000941:Antigens; D000951:Antigens, Neoplasm; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D006146:Guanidines; D012685:Sepharose; C049225:Superose 12; D019791:Guanidine