(HP-SEC) High Performance Size Exclusion Chromatography Tests

High performance size exclusion chromatography (HP-SEC) is a well-established, robust and versatile technique for the characterization and examination of protein monomers, fragments and aggregates.

In this test method, the sample is injected into a column that is continuously washed with the mobile phase. The chromatography column contains fine and porous beads of dextran, agarose, silica or polyacrylamide (stationary phase). Beads allow small species to migrate into the pores, increasing their retention in the column, while larger species migrate with the mobile phase without entering the bead pores. Thus, larger species reach the column end faster than smaller species.

After separation, various species are detected using detectors that include: ultraviolet absorption, fluorescence, refractive index, multi-angle laser light scattering, and charged aerosol detection.

The HP-SEC technique allows sizing, quantification and molecular weight determination of fragments, monomers and aggregates. The size range of this technique is defined by the pore size of the column and the method setup, eg mobile phase, flow settings, and column dimensions.

For common sample types in advanced laboratories, standardized methods are often applied with little setup effort. On top of that, experienced analytical scientists develop methods in accordance with the active ingredients, product type and development stage.

Briefly, size exclusion chromatography is a technique applied to separate proteins and other biological macromolecules on the basis of molecular size. Today, this technique, which separates molecules by size by filtration through a gel, has gained wide acceptance and is widely used because of the variety of molecular weights of proteins in biological tissues and extracts. Size exclusion chromatography is also used for many purposes, including buffer exchange (desalting), removal of non-protein contaminants (DNA, viruses), removal of protein aggregates, study of biological interactions, and protein folding (desalination). Being compatible with physiological conditions also creates an important advantage.

Our organization also provides (HP-SEC) high performance size exclusion chromatography testing services with its trained and expert staff and advanced technological equipment, among the numerous test, measurement, analysis and evaluation studies it provides for businesses in various sectors.