Glycoproteins Analysis Based on HPLC
A sample combination or analyte is pumped down a column containing chromatographic packing material (stationary phase) at high pressure using a solvent known as the mobile phase in high-performance liquid chromatography (HPLC). A stream of flowing nitrogen or helium carrier gas carries the sample. Any substance that can dissolve in a liquid and have trace amounts as low as parts per trillion can be separated and identified using HPLC. The primary benefits of HPLC are its high productivity in terms of cost parameters, great resolution, ease of use, generally high recoveries, and excellent reproducibility of repeat separations.
HPLC is a frequently employed technique in the separation and evaluation of glycoproteins and the carbohydrates they generate. A popular technique for the analysis and purification of peptides, proteins, and glycoproteins is reversed-phase HPLC, or RP-HPLC. An n-alkylsilica-based sorbent is typically included in the RP-HPLC experimental setup. With the use of contemporary equipment and columns, complex mixtures of proteins and peptides can be separated and low picomolar levels of resolved components can be collected. Separated fractions can then be utilized to analyze amino acids and carbohydrates in further detail.
