With the years of experience in the field of glycoprotein analysis, Creative Biolabs has become a world-leading custom service provider offering a full range of glycoprotein analysis services for global customers. With high-analytical sensitivity and high resolution, UHPLC/FLD/Q-TOF has been employed by our professional experts to characterize N-Glycan Profiling of a monoclonal antibody (mAb).
Glycoproteins Analysis Based on UHPLC/FLD/Q-TOF
In general, glycosylation analysis of a mAb is conducted by enzymatic deglycosylation using N-Glycosidase F (PNGase F) to cleave asparagine-linked oligosaccharides from the antibody. Then the released glycans are derivatized, fluorescently labeled, and separated using UHPLC method. An FLD is a sensitive tool for providing sensitive and quantitative analysis of the labeled glycans. However, HPLC-FLD method does not always offer positive identification of all the glycan isoforms. While Q-TOF is known to have a larger mass range and higher resolution, so lower false-positive rates. Q-TOF MS is used to definitively identify all glycan isoforms with accurate mass information. As a consequence, UHPLC/FLD/Q-TOF MS is a powerful platform for quantitative and qualitative profiling of N-glycans associated with mAbs, including minor glycoforms.
Learn more: Ultra High-Performance Liquid Chromatography with Fluorescence Detection-Quadrupole Time-of-Flight
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