HPLC Analysis

High Pressure liquid Chromatography is one of the most important tools employed by chemists in separation and purification of protein and peptide. Protein purification is the process of isolating proteins from a homogenate, which may comprise cell and tissue components, including DNA, cell membrane and other proteins.

Purification generally utilizes three properties to separate proteins. First, proteins may be purified according to their isolectric points by running them through a pH graded gel or an ion exchange column. Second, proteins can be separated according to their size or molecular weight via size exclusion chromatography or by SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) analysis. Finally, proteins may be isolated according to their unique binding properties. For example, Lactate Dehydrogenase (LDH), which binds NADH, can be purified by running it through a column comprised of beads with prosthetic NAD analog groups.

Protein purification schemes usually involve multiple separations and isolations. Furthermore, two purification methods may be employed together to increase the degree of purification. For example, in two dimensional gel electrophoresis, proteins are first separated longitudinally according to molecular weight, then laterally according to their isoelectric points.
PrimeSyn Lab offers following HPLC services:

1. Reversed-phase
2. Ion Exchange
3. Size Exclusion Chromatography