Metal affinity chromatography, applications

The ELP expression system was compared to the conventional oligohistidme fusion, which is traditionally applied for purification by immobilized metal affinity chromatography (IMAC). Both techniques were shown to have a similar yield of the recombinant protein. The temperature-triggered approach offers a fast and inexpensive nonchromatographic separation with the possibility for larger scale purification. Although the ELP expression system may not be applicable to all types of recombinant proteins, numerous examples have already been shown [40]. 

Wu, H. and Bruley, D.F, Homologous Human Blood Protein Separation Using Immobilized Metal Affinity Chromatography Protein C Separation from Prothrombin with Application to the Separation of Factor IX and Prothrombin, Biotechnol. Prog., 15, 928, 1999. 

Abbreviations for type of application IEX Ion Exchange, HIC Hydrophobic interaction chromatography, AF Affinity chromatography, RP Reverse phase, IMAC Immobilized metal affinity chromatography. 

Applications of metal affinity chromatography include separation of S -oxynitrilase from Sorghum bicolor on IDA-Cu " ion... 

The foundations for immobilized metal affinity chromatography (IMAC) were first laid in 1961 when Helferich introduced ligand-exchange chromatography [1], The modern-day usage of this technique and its practical applications as a purification tool did not emerge, however, until 1975 and the seminal work by Porath et al. [2],... 

Ndassa, Y.M. Orsi, C. Marto, J.A. Improved irtunobilized metal affinity chromatography for large-scale phosphoproteomics applications. J. Proteome Res. 2006, 5, 2789-2799. 

Holmes, L.D. Schiller, M.R. Immobilized itonflll) metal affinity chromatography for the separation of phosphory-lated macromolecules Ligands and applications. J. Liq. Chromogr. Relat. Technol. 1997, 20 (1), 123. 

A higher value application, based on functionalization of a solid particle, is the development of the stationary phase of nanoengineered analytical immobilized metal affinity chromatography columns by ATRP for separation of proteins and synthetic prion peptides. 

Metal chelate affinity chromatography finds most prominent application in the affinity purification of recombinant proteins to which a histidine tag has been attached (described later). As protein binding occurs via the histidine residues, this technique is no more inherently useful for the purification of metalloproteins than for the purification of non-metalloproteins (a common misconception, given its name). 

Advanced affinity chromatography (AAC) media are used for the adsorption of metals from ground and waste waters. The AAC technology has been used in multiple applications and is commercially available from Affinity Water Technologies, formerly Ntec Solutions, Inc. 

In 1970s, first application of metal-chelate affinity chromatography which is later named as "immobilized-metal (ion) affinity chromatography (IMAC) was perfomed. Metal-chelate chromatography technique exploits selective interactions and affinity between transition metal immobilized on a solid support (resin) via a metal chelator and amino acid residues which act as electron donors in the protein of interest [25-26]. As well as aromatic and heterocyclic compounds, proteins such as histidine, tyrosine, tyriptophane and phenylalanine posses affinity to transition metals which form complexes with compounds rich in electrons [25,27]. 

A very interesting application of affinity chromatography to the purification of halophilic enzymes was reported by Sundquist and Fahey (1988). These authors have purified the enzymes bis-y-glu-tamylcysteine reductase and dihydrolipoamide dehydrogenase from H. halohium using immobilized metal ion affinity chromatography in high-salt buffers. 

Kagedal, L. (1989) Immobilized metal ion affinity chromatography, in Protein Purification -Principles, High Resolution Methods and Applications (Janson, J.-C., Ryden, L., Eds.). VCH, Weinheim. 

A variety of micropellicular packing materials has been developed for the analysis of both small and large molecules by various HPLC modes, including ion exchange (lEC), metal interaction (MIC), reversed phase (RPC) [4], and affinity chromatography (AC) [5]. Besides analytical applications, other possible utilization of micropellicular stationary phases includes fundamental kinetic and thermodynamic studies of the retention mechanisms on a well-defined surface. Nevertheless, a relatively limited variety of micropellicular... 

Many of the techniques used to enrich PTMs at the protein level are applicable at the peptide level. A popular method for enrichment of phosphopeptides is to use an immobilized metal affinity column. The molecular basis for the enrichment is the phosphate affinity to transition metal ions, such as copper, nickel, cobalt, iron, aluminum, gallium, and zinc (142). A comprehensive source for phosphopeptide enrichment strategies can be found in a recent review (143). Lectin-based chromatography is used to enrich for glycopep-tides (144). Lectins are proteins that have an affinity for carbohydrates. With multiple available enrichment options, choosing the right one for the experiment is important. For determining which proteins have a certain PTM, protein-level enrichment can be performed. However, if a type of PTM is of... 

These biopolymers can be used for the immobilization of metal ions not only with the final objective of metal recovery (and subsequent valorization by desorption or chemical/thermal destmction of the polymer matrix) but also for elaborating new materials or designing new applications. Depending on the metal immobilized on the biopolymer, it is possible to design new sorbents (immobilization of iron on alginate [119], of molybdate on chitosan [59], for As(V) removal, of silver on chitosan for pesticide removal [120]), supports for affinity chromatography [121], antimicrobial material [122], drug release material [123], neutron capture therapy [124], and photoluminescent materials [125]. These are only a few... 

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