The characteristic analysis of sugar chains bound to proteins is essential for the development of antibody drugs which has recently received attention. Various derivatization reagents are used for high sensitivity analysis of sugar chains with HPLC, and 2-aminobenzoic acid (2-AA) is one of typical sugar derivatization reagents. In this application, 2-AA Labeled sugar chain (glucose homopolymer) was separated usign a polymer-based amino column, under HILIC mode. Derivatization with 2 – AA allows the use of a fluorescence detector for sugar chain analysis, and G1 (glucose) can be detected in amol levels.
Sample : 50μL
2-AA Labeled glucose homopolymer ladder (2-AA Labeled dextran hydrolysate ladder)
10nM (G1 conc.)
Column : Shodex Asahipak NH2P-50 4E (4.6mmI.D. x 250mm) Eluent : (A); 2% CH3COOH in CH3CN (B); 5% CH3COOH + 3% Triethylamine in H2O (B%) 30% to 95% (2 to 82min) Flow rate : 1.0mL/min Detector : Fluorescence (Ex.350nm, Em.425nm) Column temp. : 40℃
