Protein Selectivity with Immobilized Metal Ion-tacn Sorbents: Chromatographic Studies with Human Serum Proteins and Several Other Globular Proteins

Separation of hexahistidine fusion proteins with immobilized metal ion affinity chromatographic (IMAC) sorbents derived from MN+‐tacn and its derivatives

“…Moreover, these trends follow the known binding behavior of Ndonor groups, such as the nitrogen lone pair electrons of the imidazole moiety of histidine residues, which show a high propensity for coordinative interactions with Cu 2þ -or Ni 2þ ions. Related findings with human serum protein, myoglobins, cytochrome cs, lysozymes, and synthetic peptides also follow similar trends with these new IMAC ligands (Graham et al, 2008;Jiang et al, 1998).…”

“…The present investigation extends our previous studies (Graham et al, 2008;Jiang et al, 1998) on the application of tacn and its derivatives as chelating ligands, to which different soft, borderline or hard metal ions have been complexed, for use in IMAC. In this study, the comparative chromatographic properties of the im-M nþ -tacn, im-M nþ -dtne and im-M nþ -dtnp IMAC systems have been examined with the recombinant fusion protein Schistosoma japonicum glutathione S-transferase-Saccharomyces cerevisiae mitochondrial ATP synthase d-subunit containing a C-terminal hexahistidine tag (GST-dATPase-His 6 ).…”

“…The potential of IMAC sorbents derived from the macrocycle, 1,4,7-triazacyclononane (tacn), has been documented in IMAC applications related to the purification of human serum proteins and other wild-type proteins (Graham et al, 2008;Jiang et al, 1998) from biological extracts. Compared to other tri-or tetradentating chelating ligands, tacn and its derivatives have much larger stability constants for many borderline metal ions, with values typically higher than IDA by at least 4-10 orders of magnitude, that is, in the range of 16 log b 30 (Haidar et al, 1997;Spiccia et al, 1997;Tanaka et al, 1977;Zhang et al, 1995).…”

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

“…Tacn-based ligands exhibit very low toxicity indices and can be easily immobilized onto support materials via oxirane or other chemistries [13][14][15][16]. With histidine-containing globular native proteins, such as myoglobin or cytochrome c, binding capacities in the range of 15-25 mg protein/mL gel have been observed with these tacn-based adsorbents [13,14].…”

High‐Performance Liquid Chromatography of Biological Macromolecules

“…In connection with these borderline metal ions, the tacn-based sorbents showed a new selectivity toward proteins, compared to previous chelating groups based on IDA, NTA, or OPS. 523 In two recent studies, they make use of 2,6-diaminomethylpyridine 524 and N-(2-pyridylmethyl)aminoacetate 525 utility, commercial availability is a problem with all the less-common chelating sorbents. Therefore, IDA remains the most widely used sorbent functionality and is available from several sources.…”