Analytical Separations of Proteins and Peptides Using Immobilized Metal Affinity Chromatography

摘要

Immobilized metal affinity chromatography (IMAC) is a well-established protein separation technique experiencing expanded use based on new, high-performance analytical phases. IMAC is commonly used in capture/ release cartridges for enriching peptidesand proteins of a certain class (i.e., surface exposed histidine [His] or phosphopeptides/phosphoproteins) or His-tagged recombinant protein purification. Due to die relatively low resolution of these cartridges, fractions collected from the cartridgescan still be quite complex, and further purification is generally required. To address this limitation, a reusable, high-resolution IMAC column that can be used for high-performance (1-D and 2-D) LC separations has been developed.