1997
DOI: 10.1021/ac960882d
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Characterization by Potentiometric Procedures of the Acid−Base and Metal Binding Properties of Two New Classes of Immobilized Metal Ion Affinity Adsorbents Developed for Protein Purification

Abstract: The acid-base protonation constants of two recently introduced chelating ligands for protein purification, O-phosphoserine and 8-hydroxyquinoline immobilized onto Sepharose CL-4B, and the stability constants of their derived immobilized metal ion chelate complexes have been determined by potentiometric methods. The data confirm that immobilization thermodynamically constrains the ligands, with the electron-withdrawing characteristics of the group linking the ligand to the support material affecting the magnitu… Show more

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Cited by 22 publications
(12 citation statements)
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“…While the (c) process involves the formation of a coordination bond between imidazole and the immobilized metal ion and a nonspecific interaction, the nonspecific interaction should not be neglected as the electrostatic repulsion between protein and adsorbents under no salt present. As can be seen from the literature (21,(31)(32)(33), the negative charge of the CS-IDA-Fe(III) is higher than that of CS-IDA-Cu(II). Therefore, the heat required to overcome the nonspecific interaction (electrostatic repulsion) of Fe(III) is higher than of Cu(II) at pH 8.5, where lysozyme is negatively charged.…”
Section: Metal Effectsmentioning
confidence: 80%
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“…While the (c) process involves the formation of a coordination bond between imidazole and the immobilized metal ion and a nonspecific interaction, the nonspecific interaction should not be neglected as the electrostatic repulsion between protein and adsorbents under no salt present. As can be seen from the literature (21,(31)(32)(33), the negative charge of the CS-IDA-Fe(III) is higher than that of CS-IDA-Cu(II). Therefore, the heat required to overcome the nonspecific interaction (electrostatic repulsion) of Fe(III) is higher than of Cu(II) at pH 8.5, where lysozyme is negatively charged.…”
Section: Metal Effectsmentioning
confidence: 80%
“…The nonspecific interaction can be affected by the charge density and binding capacity of metal ions on the gel. The literature has also reported (20,(31)(32)(33) that the negative charge on the Sepharose gel was Fe(III) higher than Cu(II).…”
Section: Introductionmentioning
confidence: 95%
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“…Advantage can also be taken of the relative metal ion stability constant (as the average logb value) in selecting such IMAC systems. Both potentiometric titration and isothermal titration microcalorimetry are particularly useful in characterising different types of soft/borderline/hard metal ion systems with immobilised chelating ligands system, and enable the respective hydrolytic complexes that form with different immobilized metal ion chelating complexes (IMCC's) to be properly defined (Zachariou et al, , 1997Hearn, 2000). Indicative of the potential of these approaches have been various investigations with synthetic peptides (Nakagawa et al, 1988;Porath, 1988a,b Kronina et al, 1999, as well as studies with a-interferon -related peptides (Sulkowski 1989), whilst the elegant exploratory studies related to bovine growth hormone (Suh et al, 1991) has been elaborated above.…”
Section: Histidine Tags For the Separation Of Recombinant Proteins Bymentioning
confidence: 99%
“…Only limited studies have concentrated on the acid-base and metal binding properties of new and traditional chelating adsorbents and on changes in the physicochemical properties of immobilized metal chelate complexes. [21][22][23][24] Because of the weak interaction between vinyl imidazole and Cu 21 ions in the solid IMA chromatography matrixes, no data exist on complex stability and complex stoichiometry in literatures. Until now, the strength of the Cu 21 -vinyl imidazole complex has been determined only in the water-soluble linear poly (1-vinyl imidazole) systems.…”
Section: Introductionmentioning
confidence: 99%