Polymer-shielded dye-affinity chromatography

Mattìasson, Bo; IYu, Galaev; Garg, Nandita

doi:10.1002/(sici)1099-1352(199634/12)9:5/6<509::aid-jmr293>3.0.co;2-y

Cited by 15 publications

(2 citation statements)

References 17 publications

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“…Chemistries for immobilization of dye ligands via epoxide-activated spacers instead of the conventional coupling to surface hydroxyls were recently investigated by Scoble and Scopes, 385 who showed that the epoxide immobilization route gave materials with considerably higher ligand density. Mattiasson et al 386 addressed the nonspecific interactions of dye affinity sorbents by shielding the ligands with layers of nonionic, water-soluble polymers such as poly(vinylpyrrolidone) or poly(vinyl alcohol). The same authors also reviewed new developments in dye affinity for bioprocessing, dealing specifically with new dyes and conditions for the protein -ligand attachment and detachment steps.…”

Section: Enzyme -Inhibitor Complexesmentioning

confidence: 99%

High‐Performance Liquid Chromatography of Biological Macromolecules

Irgum¹

2000

Encyclopedia of Analytical Chemistry

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Section: Enzyme -Inhibitor Complexesmentioning

confidence: 99%

High‐Performance Liquid Chromatography of Biological Macromolecules

Irgum¹

2000

Encyclopedia of Analytical Chemistry

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“…However, even with an ideal ligand density, it is still possible for weak specific interactions to occur between CB and contaminant proteins, which will potentially decrease the purity of the final product. In order to ameliorate this problem, Mattiasson et al [11] introduced the concept of 'polymer shielding', whereby one loads on to the affinity column a polymer that is less strongly bound than the product molecule but more strongly bound than contaminants, thereby 'shielding' the column from non-specific adsorbates. Here we assess the potential for an intrinsic shielding approach, whereby BSA, a protein known to show a weak interaction with CB, is co-immobilized to the support.…”

Section: Introductionmentioning

confidence: 99%

An intrinsically shielded hydrogel for the adsorptive recovery of lysozyme

Wang

Zhang

Eisenthal

et al. 2006

Biotech and App Biochem

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The present paper addresses the selective recovery of lysozyme from egg white using CM-dextran (carboxymethyldextran)-based hydrogels containing Cibacron Blue as an affinity ligand and co-immobilized BSA intended to act as a shielding agent to reduce non-specific adsorption. Initial studies using pure lysozyme were conducted that indicated that the adsorption capacity increased with ligand density and that adsorption was well described by a Langmuir-type isotherm. The inclusion of BSA as a putative shielding agent did not decrease the adsorption capacity for lysozyme in single-adsorbate experiments. To assess the effectiveness of the shielding strategy, subsequent experiments were conducted with both defined lysozyme/ovalbumin mixtures and hen's-egg white. From these studies, the optimal operating conditions for lysozyme recovery have been determined. These include: optimal initial egg-white concentration [a 10% (v/v) solution of native egg white in the chosen buffer], affinity-ligand density (1.86 mM) and ligand-to-shielding-agent ratio (4:1). The purity of lysozyme obtained from egg white was improved from 69% with a non-shielded hydrogel to 94% with an intrinsically shielded hydrogel. Finally, the possibility of using a protein, rather than dextran-backbone-based, hydrogel was investigated. It was found that BSA could take the place of CM-dextran as the gel backbone in a simplified synthesis, producing a gel which also proved effective for lysozyme recovery with a 30% lysozyme in egg-white solution purified to approx. 92% in a single adsorption-desorption cycle.

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