Úloha glykokonjugátů v buněčné signalizaci - CEACAMI a Fcε R1

• Main Author: Dudková, Lenka
• Other Authors: Šmíd, František
• Format: Doctoral Thesis
• Language: Czech
• Published: 2006
• Online Access: http://www.nusl.cz/ntk/nusl-270081

Carcinoembryonic cell adhesion molecule 1 (CEACAM1) is human membrane glycoprotein. The purpose of the first part of this thesis was to isolate CEACAM1 glycoprotein from bile and characterise its purity and recovery that have not been described before. Affinity chromatography of CEACAM1 on hydrazide-activated cellulose with immobilized monoclonal anti-CEA F34-187 antibodies is described. The method is based on the oxidation of the immunoglobulin carbohydrate component located on the Fc fragment of antibody by periodate and oriented bounding to hydrazide-activated matrix. Crude protein fraction from bile was applied onto the affinity column and CEACAM1 was eluted with 6 M guanidine-HCl. A single immunopositive 85 kDa band of CEACAM1 was detected on the western blot with anti-CEA antibody. Probably due to the high glycosylation of CEACAM1 any common method of protein staining was not applicable except staining with lectin.