Abstract
Immobilized metal ion affinity chromatography (IMAC) (1,2) is also referred to as metal chelate chromatography, metal ion interaction chromatography, and ligand-exchange chromatography. We view this affinity separation technique as an intermediate between highly specific, high-affinity bioaffinity separation methods, and wider spectrum, low-specificity adsorption methods, such as ion exchange. The IMAC stationary phases are designed to chelate certain metal tons that have selectivity for specific groups (e.g., His residues) in peptides (e.g., 3–7) and on protein surfaces (8–13). The number of stationary phases that can be synthesized for efficient chelation of metal ions is unlimited, but the critical consideration is that there must be enough exposure of the metal ion to interact with the proteins, preferably in a biospecific manner. Several examples are presented in Fig. 1. The challenge to produce new immobilized chelating groups, including protein surface metal-binding domains (14,15) is being explored continuously (16). Table 1 presents a list of published procedures for the synthesis and use of stationary phases with immobilized chelating groups. This is by no means exhaustive, and is intended only to give an idea of the scope and versatility of IMAC.
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© 1996 Humana Press Inc.
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Yip, TT., Hutchens, T.W. (1996). Immobilized Metal Ion Affinity Chromatography. In: Doonan, S. (eds) Protein Purification Protocols. Methods in Molecular Biology™, vol 59. Humana Press. https://doi.org/10.1385/0-89603-336-8:197
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DOI: https://doi.org/10.1385/0-89603-336-8:197
Publisher Name: Humana Press
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