High-performance liquid chromatography (HPLC) is a sensitive analytical technique that is widely used for the isolation and analysis of both glycoproteins and their derived carbohydrates. Based on this technique, Creative Biolabs is committed to offering a wide range of glycoprotein analysis service for customers all over the world.
HPLC is a form of column chromatography that pumps a sample mixture or analyte in a solvent (called the mobile phase) at high pressure through a column with chromatographic packing material (stationary phase). The sample is carried by a moving carrier gas stream of helium or nitrogen. HPLC has the ability to separate and identify compounds that are present in any sample that can be dissolved in a liquid in trace concentrations as low as parts per trillion. The main advantages of HPLC include excellent resolution, ease of use, the generally high recoveries, the excellent reproducibility of repetitive separations, and the high productivity in terms of cost parameters.
HPLC is also one of the most commonly used methods for the isolation and analysis of both glycoproteins and their derived carbohydrates. Reversed-phase HPLC (RP-HPLC) has become a commonly used method for the analysis and purification of peptides, proteins, and glycoproteins. The RP-HPLC experimental system usually includes an n-alkylsilica-based sorbent. Complex mixtures of peptides and proteins can be separated and low picomolar amounts of resolved components can be collected by using modern instrumentation and columns. Subsequently, separated fractions can be used for further analysis of carbohydrates and amino acids. Besides, a new HPLC method, HPLC-pulsed amperometric detection (PAD), which bypasses the derivatization steps by using pulsed electrochemical detection on gold electrodes, has been developed. HPLC-PAD has been used successfully in resolving and quantitating the constituent monosaccharides released by acidic hydrolysis of glycan chains and for resolving N-linked oligosaccharides separated by enzyme digestion.
Fig. 1 Preparative HPLC system.
Creative Biolabs has successfully developed an advanced and sensitive platform for glycoprotein/glycopeptide analysis based on HPLC technique. Based on RP-HPLC, complex mixtures of glycoproteins can be separated efficiently and the separated carbohydrates and amino acids can be analyzed by combined with other detection technologies. Based on HPLC-PAD, monosaccharides released by acidic hydrolysis of glycan chains are quantitatively analyzed. Besides, we also provide a custom service for global clients to meet their special purposes.
Professional scientists in Creative Biolabs can help our client to design the optimal analytical protocol and attain their interested experimental data to meet their special project requirement. We have established a powerful HPLC-based analytical platform which is applied for Glycan Profiling, Glycomic Profiling, and Glycosylation Site Mapping. If you are interested in our HPLC analysis services, please feel free to contact us for more details.