A rapid colorimetric assay for carcinoembryonic antigen (CEA)-mediated cell adhesion and analysis of CEA domains involved in the adhesion.
A colorimetric microadhesion assay that allows the quantitative determination of carcinoembryonic antigen (CEA)-mediated homophilic cell adhesion to CEA immobilized on 96-well polyvinyl chloride plates is described. Chinese hamster ovary (CHO) cells transfected with a full-length CEA cDNA were used as indicator cells. After dislodging nonadherent cells, specifically bound cells were quantified by a colorimetric analysis based on the ability of live cells to reduce the dye 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to a blue formazan product. The domains of CEA responsible for the homophilic cell adhesion were analyzed by inhibition assays using anti-CEA monoclonal antibodies whose reactive domains were already known. The involvement of domain N and possibly subdomain A3 of CEA in the homophilic cell adhesion has been suggested.
Kuwahara
M
M
First Department of Biochemistry, School of Medicine, Fukuoka University, Japan.
Kuroki
M
M
Haruno
M
M
Murakami
M
M
Arakawa
F
F
Oikawa
S
S
Nakazato
H
H
Matsuoka
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001665:Binding Sites / Q000187:drug effects; D016466:CHO Cells / Q000378:metabolism; D002272:Carcinoembryonic Antigen / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D002448:Cell Adhesion; D002452:Cell Count; D002470:Cell Survival; D002478:Cells, Cultured; D003124:Colorimetry / Q000379:methods; D006224:Cricetinae; D004273:DNA, Neoplasm / Q000235:genetics; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D013778:Tetrazolium Salts; D013844:Thiazoles; D014162:Transfection / Q000235:genetics
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D004273:DNA, Neoplasm; D000939:Epitopes; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue