Polymer displacement in dye-affinity chromatography

Galaev, Igor Yu.; Arvidsson, Per-Ola; Mattìasson, Bo

doi:10.1016/0021-9673(95)00486-6

Cited by 14 publications

(12 citation statements)

References 36 publications

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“…Using difference spectroscopy and fitting the data to a simple mathematical model, we were able to determine the binding constants of some water-soluble polymers with the dye-ligands in solution (Galaev and Mattiasson, 1994;Galaev et al, 1994aGalaev et al, , 1995Garg et al, 1997). Non-charged weakly hydrophobic polymers such as poly(N-vinyl pyrrolidone) (PVP) interact with Cibacron Blue 3GA in solution with binding constants of 2-6 mM depending on the molecular weight of the polymer.…”

Section: Resultsmentioning

confidence: 99%

“…To the best of our knowledge, polymer displacement in dye-ligand chromatography was initiated in our laboratory (Galaev and Mattiasson, 1993a;Galaev et al, 1994bGalaev et al, , 1995. The main objective of this paper is to present the state of the art in using polymers for displacement of proteins from dye-ligand matrices and to discuss the results obtained with the combination of some data on polymer-dye interactions.…”

Section: Introductionmentioning

confidence: 99%

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Protein displacement in dye–ligand chromatography using neutral and charged polymers

1998

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Displacement chromatography was demonstrated to perform separations efficiently under mass-overloaded conditions, offering advantages such as increased product recovery and purity, superior resolving power, and concentration and purification in a single processing step. The use of water-soluble polymers for protein displacement in dye-ligand chromatography was initiated in our laboratory. The polymers for displacement were selected using differences spectroscopy to monitor their interactions with a dye-ligand in solution. Non-charged polymers such as poly(N-vinyl pyrrolidone) and poly(N-vinyl caprolactam) efficiently displaced lactate dehydrogenase from porcine muscle from a Blue Sepahrose column. The latter polymer, being thermosensitive, could be easily removed from the eluate and recovered by precipitation at 45 degrees C and low-speed centrifugation. The positively charged polymer poly(ethylene imine) proved to be an even more efficient displacer. The dye-ligand column could be regenerated after application of displacer either by washing with a solution of the soluble ligand Cibacron Blue (in the case of non-charged polymers) or by washing with highly alkaline solutions containing polyanions (in the case of poly(ethylene imine)) The latter formed a soluble complex with poly(ethylene imine) and stripped the column from the polymer.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Protein displacement in dye–ligand chromatography using neutral and charged polymers

1998

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“…The advantage of using an aqueous solution of PVCL was that above its cloud point of 40°C, it separates into two phases, an upper water-rich and a lower polymer-rich phase, so if the eluate containing PVCL was heated to 45OC the enzyme was recovered in the water-rich phase in a eluent-free form, and regenerated PVCL could be used repeatedly. Poly(ethy1ene imine) proved to be an efficient displacer of lactate dehydrogenase bound to Cibacron Blue F3GA-Sepharose or Procion Red HE3B-Sepharose (Galaev et al, 1995).…”

Section: Recovery Of Bound Proteinmentioning

confidence: 98%

Dye-affinity techniques for bioprocessing: Recent developments

1996

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Textile or triazine dyes play an important role as affinity ligands in protein purification. Each step of the protein purification protocol can be divided into three stages, partitioning between two phases, separation of these phases and recovery of the target protein from the enriched phase. Now developments in dye-affinity techniques are discussed emphasizing the innovations in all three stages of the protein purification process. Dye-affinity chromatography has become a routine step in protein purification. New dyes have been developed and used successfully in both traditional chromatographic mode and new modes like affinity precipitation, polymer aqueous two-phase partitioning or expanded bed chromatography. The specificity of dye techniques has been increased by both purposeful designing of new dyes and decreasing non-specific protein-dye interactions with polymer shielding. One can envisage further development and ramification of dye-affinity techniques in protein purification.

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“…It is now well established that triazine dyes have shown a striking aptitude for protein purification, stemming from their chemical structure having remarkable similarities to the active sites of many enzymes. Thus, these smaller, cheaper, and much more stable compounds can often be used to replace delicate biological ligands for affinity chromatography (Alderton et al, 1994;Costaferreira and Duarte, 1991;Galaev et al, 1995;Labrou and Clonis, 1995;Ruthven and Ching, 1989;Tuncel et al, 1993). Although literature describing the successful application of dyeaffinity chromatography is plentiful, this technique is still not being exploited to its deserved extent in industry, even despite increasing economic pressures on manufacturers to produce protein-based products cheaply.…”

Section: Introductionmentioning

confidence: 99%

A new approach to continuous counter-current protein chromatography: Direct purification of malate dehydrogenase from aSaccharomyces cerevisiae homogenate as a model system

Owen

McCreath

Chase

1997

Biotechnol. Bioeng.

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A novel technique for protein chromatography has been developed, which can be used to extract proteins from particulate‐containing solutions (such as fermentation broths or preparations of disrupted cells) on a continuous basis, and delivers clarified streams of purified product. Adsorbents deployed in this type of contactor are based on PVA‐coated perfluorocarbons derivitized with affinity ligands such as triazine dyes. In this article, we describe the application of this equipment for the continuous purification of malate dehydrogenase from an unclarified homogenate of Saccharomyces cerevisiae, using a Procion Red HE‐7B‐derivitized adsorbent. Although operating conditions were not optimized to produce a product of maximized purification factor, concentration, and yield, we have shown that MDH can be purified continuously in 78% yield at a rate of 70 U/min, with a purification factor of approximately 10. This corresponds to specific productivity of approximately 0.35 U/min per milliliter of settled adsorbent, a higher specific productivity than was feasible with the same adsorbent using expanded bed adsorption (EBA). © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 427–441, 1997.

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Polymer displacement in dye-affinity chromatography

Cited by 14 publications

References 36 publications

Protein displacement in dye–ligand chromatography using neutral and charged polymers

Protein displacement in dye–ligand chromatography using neutral and charged polymers

Dye-affinity techniques for bioprocessing: Recent developments

A new approach to continuous counter-current protein chromatography: Direct purification of malate dehydrogenase from aSaccharomyces cerevisiae homogenate as a model system

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