High-throughput platform introduced in Genos enables fast and robust analysis of protein glycosylation.
UPLC (Ultra Performance Liquid Chromatography) glycosylation analysis:
Ultra Performance Liquid Chromatography is generally accepted as the gold standard for quantitation of a wide range of glycan structures. Enzymatically released glycans from proteins are labelled with 2-aminobenzamide, a fluorescent dye which in combination with a fluorescent detector enables high sensitivity of the system. Glycans are separated using Hydrophilic interaction chromatography (HILIC) based on charge, size, and polarity of glycan. UPLC analysis is a sensitive method with high resolution, reduced analysis time and cost-per-sample. It is a reliable and robust quantitative method proven to provide quality data which makes it the method of choice for large studies (publications).
Liquid chromatography – mass spectrometry (LC-MS) analysis of glycopeptides:
LC-MS is another powerful tool for the structural characterization of glycans due to its sensitivity, small sample requirement and ability to analyze complex glycan mixtures. In addition to the characterization of the glycan sequence, MS provides branching patterns, substituent location, and quantitative information. In this approach glycans are still attached to a peptide backbone which preserves the information about the position of the glycosylation site in the studied protein. Genos has introduced and validated a fast nano-reverse phase-LC-ESI-MS method developed for high-throughput analysis of glycopeptides. The advantage of softer ionization techniques like Electro spray ionization (ESI) enables the analysis of intact ions of high molecular mass and provides high sensitivity.