Use of phage display methods to identify heptapeptide sequences for use as affinity purification ‘tags’ with novel chelating ligands in immobilized metal ion affinity chromatography

Mooney, Jane Therese; Fredericks, Dale Patricia; Hearn, Milton Thomas William

doi:10.1016/j.chroma.2010.10.113

Cited by 30 publications

(23 citation statements)

References 41 publications

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“…Since these ligands are the only nitrogen-containing component of the adsorbents, the extent of ligand immobilization could be readily determined via nitrogen analysis ( Table 1). The ligand densities achieved are similar to those reported for other IMAC adsorbents [6][7][8][9][10][11]. Under the same immobilization conditions, a higher surface coverage was observed for the immobilized tacn adsorbent.…”

Section: Preparation and Characterization Of Imac Adsorbentssupporting

confidence: 88%

“…Acetonitrile was dried over 4Å molecular sieves. The synthesis of tacn and tacn 2 butane was based on published methods [8][9][10][11]. Chicken egg white lysozyme (HEWL) (3 × crystallized, dialyzed and lyophilized; 95% minimum protein), horse skeletal muscle myoglobin (HMYO) (95% minimum protein) and horse heart cytochrome c (HHCC) (95% minimum protein) were all purchased from Sigma Chemical Co. (St. Louis, MO, USA).…”

Section: Materials and Reagentsmentioning

confidence: 99%

“…The binding experiments were conducted under batch equilibrium conditions [8][9][10][11] with the extent of protein bound determined either by direct spectrophotometric measurement or by a dye-binding assay [23]. The extent of protein adsorption to both the Cu 2+ -loaded and metal-ion-free gels was measured with acetate (pH 5.0), phosphate (pH 7.0) and borate (pH 9.0) buffers containing NaCl at a concentration ranging from 0, 0.5 or 1 M.…”

Section: Binding Studies With Several Globular Proteins To the Imac Amentioning

confidence: 99%

“…Previously, we have described the application of such alternative types of chelating ligands for use in the IMAC separation of proteins. In particular, we have documented the chromatographic behavior of immobilized copper(II) complexes of o-, m-and pxylylene bridged bis(1,4,7-triazacyclononane) macrocycles in the IMAC separation of proteins [9], as well as the utility of the immobilized borderline metal ion complexes of Cu 2+ , Ni 2+ , Zn 2+ , Mn 2+ , Co 2+ and Cr 3+ with 1,4,7-triazacyclononane (tacn) for the purification of hexahistidine tagged [10] and other more specifically tagged [11] recombinant proteins. In this paper we extend these studies and describe investigations into the binding characteristics of several proteins with a new IMAC adsorbent, namely Cu 2+ -loaded 1,4-bis(1,4,7-triazacyclononan-1-yl)butane (tacn 2 butane) functionalized sepharose CL-4B.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of the binding behavior of several histidine-containing proteins with immobilized copper(II) complexes of 1,4,7-triazacyclononane and 1,4-bis(1,4,7-triazacyclononan-1-yl)butane

Graham

Spiccia

Hearn

2011

Journal of Chromatography B

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Section: Preparation and Characterization Of Imac Adsorbentssupporting

confidence: 88%

Section: Materials and Reagentsmentioning

confidence: 99%

Section: Binding Studies With Several Globular Proteins To the Imac Amentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Comparison of the binding behavior of several histidine-containing proteins with immobilized copper(II) complexes of 1,4,7-triazacyclononane and 1,4-bis(1,4,7-triazacyclononan-1-yl)butane

Graham

Spiccia

Hearn

2011

Journal of Chromatography B

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“…Recently, we described a new family of histidine-containing tags that display strong affinity for tacn-based IMAC adsorbents [19,27]. The ability to completely remove these new tags was also a major factor in their design, a process often required for the production of recombinant proteins intended for structure-function studies or use in therapeutic applications [28][29][30][31][32].…”

Section: Resultsmentioning

confidence: 99%

Changed loading conditions and lysate composition improve the purity of tagged recombinant proteins with tacn‐based IMAC adsorbents

Zhang

Fredericks

Longford

et al. 2014

Biotechnology Journal

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These investigations were designed to improve capture efficiency and selectivity in the immobilized metal ion affinity chromatographic (IMAC) purification of tagged recombinant proteins expressed in Escherichia coli cells, utilizing an alternative and novel class of immobilized metal binding ligands. The impact of loading conditions and lysate composition on the IMAC purification of NT1A- or His6 -tagged green fluorescent protein (GFP), using the ligands 1,4,7-triazacyclononane (tacn) and bis(1,4,7-triazacyclononyl)propane (dtnp), charged with Cu(2+) ions, has thus been explored. These findings were compared to the performance of a commercial adsorbent, IMAC Sepharose™ 6 FF, similarly charged with Cu(2+) ions. With the same loading, wash and elution protocols, the tacn- and dtnp-derived adsorbents showed higher selectivity in terms of removal of E. coli host cell proteins than the commercial adsorbent, while low molecular weight components in the crude lysate had a higher impact on the binding capacities of tacn- and dtnp-derived adsorbents. This effect of lysate composition could be reduced through osmotic shock treatment of the E. coli cells prior to lysis. Additionally, the protein-binding capacities of the tacn-based resins were enhanced by increasing their ligand densities. Because both the tacn- and the dtnp-derived IMAC adsorbents exhibit very high metal ion stability constants, under the chromatographic conditions examined, they could be used several times without re-charging with Cu(2+) ions. The results of these studies thus expand the general application scope of tacn-based IMAC resins for use in the capture and purification of tagged recombinant proteins.

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Metalloprotein Design and Engineering

Chakraborty

Hosseinzadeh

2014

Encyclopedia of Inorganic and Bioinorganic Chemistry

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This article covers recent advances in metalloprotein design, focusing on different approaches to their design. Impressive progress has been made in designing metal‐binding sites in peptides, de novo designed proteins, and native protein scaffolds. The design approach can be rational or combinatorial. Under rational design, redesigning an existing metal‐binding site to a new site with dramatically different structure and function complements well the design of new metal‐binding sites by revealing the role of specific residues responsible for a particular structural or functional feature of the metal‐binding site of interest. To create a new metal‐binding site, several approaches have been used, including design based on structural homology, by inspection, using automated computer search algorithms, or combination of the above approaches. In addition, a modular approach involving transplanting a conserved structural unit from one protein into another has also been shown to be effective. Design through combinatorial and evolutionary methods has also been successful, as it requires little prior knowledge of the protein structure. Finally, introducing unnatural amino acids or nonnative metal ions/prosthetic groups to expand the repertoires of metalloproteins has been demonstrated. Successful examples of each of the approaches are given, advantages and disadvantages of the approaches are discussed, and the outlook for future research is also presented.

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Use of phage display methods to identify heptapeptide sequences for use as affinity purification ‘tags’ with novel chelating ligands in immobilized metal ion affinity chromatography

Cited by 30 publications

References 41 publications

Comparison of the binding behavior of several histidine-containing proteins with immobilized copper(II) complexes of 1,4,7-triazacyclononane and 1,4-bis(1,4,7-triazacyclononan-1-yl)butane

Comparison of the binding behavior of several histidine-containing proteins with immobilized copper(II) complexes of 1,4,7-triazacyclononane and 1,4-bis(1,4,7-triazacyclononan-1-yl)butane

Changed loading conditions and lysate composition improve the purity of tagged recombinant proteins with tacn‐based IMAC adsorbents

Metalloprotein Design and Engineering

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