Isolation and purification of endo-polygalacturonase by affinity chromatography in a fluidized bed reactor

Somers, W.; Reit, K. Van 't; Rozie, H.; Rombouts, F.M.; Visser, Joost

doi:10.1016/0300-9467(89)80046-2

Cited by 39 publications

(19 citation statements)

References 13 publications

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“…Therefore, there may be a need to recycle the feedstream through the bed to obtain a high degree of product capture. 31 This process of recycling is time consuming and reduces the productivity of this purification procedure. A number of methods can be used to minimize mixing of adsorbent beads in a fluidized bed, including the division of the bed into several sections by the insertion of baffles6 or the use of magnetically stabilized fluidized beds."…”

Section: Ntrodu Ctlo Nmentioning

confidence: 99%

Development of an expanded bed technique for an affinity purification of G6PDH from unclarified yeast cell homogenates

Chang

McCreath

Chaset

1995

Biotech & Bioengineering

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The use of an expanded bed of STREAMLINE Red H-7B for the purification of the intracellular glycolytic enzyme glucose 6-phosphate dehydrogenase (G6PDH) directly from untreated preparations of disrupted yeast cells has been investigated. Small-scale experiments, carried out in packed beds, have shown that the optimal pH for adsorption is 6.0 and have enabled optimization of elution conditions using a series of eluents. The dynamic capacity of the adsorbent for G6PDH was determined in a small expanded bed to be 28 units/mL. These results were used to develop a preparative scale separation of G6PDH in a STREAMLINE 50 expanded bed column. G6PDH was purified directly from an unclarified yeast homogenate in 99% yield with an average purification factor in the eluted fraction of 103. Cleaning-in-place (CIP) procedures using 0.5 M NaOH and 4M urea in 60% (v/v) ethanol have demonstrated that the adsorbent can be regenerated with no loss of adsorption capacity of alteration of bed expansion characteristics after many cycles of operation.

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Section: Ntrodu Ctlo Nmentioning

confidence: 99%

Development of an expanded bed technique for an affinity purification of G6PDH from unclarified yeast cell homogenates

Chang

McCreath

Chaset

1995

Biotech & Bioengineering

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“…However, feedstocks containing particulates can be processed directly using fluidized or expanded bed adsorption procedures which are able to operate efficiently in the presence of particulates. 5,[8][9][10][11][12]22,23,27,37,44,48 G enerally, the behavior of expanded beds is different from conventional liquid-solid fluidized beds where both liquid and adsorbent phases are well mixed, which results in poor adsorption performance and low prod~ctivity. '~ In our work, a stable bed of adsorbent beads can be easily established without need to stabilize a fluidized bed using magnetic Previous work has described the operating principles of expanded bed ad~orption.~.…”

Section: Introductionmentioning

confidence: 99%

Development of operating conditions for protein purification using expanded bed techniques: The effect of the degree of bed expansion on adsorption performance

Chang

Chase

2000

Biotechnol. Bioeng.

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A strategy for the optimization of an expanded bed adsorption process has been developed by studying a model system involving the adsorption of lysozyme onto the adsorbent STREAMLINE SP. The hydrodynamic and adsorption properties of this ion exchange adsorbent in a variety of viscosities of feedstocks have been compared by analyzing bed expansion characteristics, liquid phase dispersion characteristics, equilibrium adsorption isotherms, and mass transfer characteristics. Additionally, the influences of the degree of bed expansion on adsorption performance have been investigated by frontal analysis. In these experiments, viscous feedstocks were simulated by the inclusion of glycerol in the adsorption buffers. Breakthrough curves for lysozyme were characterized and compared in terms of overall purification processing time and productivity. On the basis of these results, the relative productivities of different operating modes with the same process liquid were found to be almost the same. However, the processing time for each purification cycle decreased with increasing velocity of process liquid. It is demonstrated that an adsorption process carried out at a constant degree of bed expansion (twice its settled bed height, corresponding to bed voidage of 0.7) is more efficient, when characterized by the apparent dynamic binding capacity, than operation at a constant liquid velocity of 300 cm/h. These results have significant implications on the design and operation of the expanded bed adsorption procedures. The advantages and problems encountered in the use of expanded bed techniques for the direct extraction of proteins from unclarified feedstocks are also discussed. © 1996 John Wiley & Sons, Inc.

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“…This modified technique is a combination of three modes of operations, which involved fluidized, suspended and expanded bed chromatographies. In fluidized bed purification, recycling of feedstock has been employed in order to increase the target protein binding and it is only feasible to use for stable protein purification [44]. As this method is rather laborious and time-consuming [44], therefore, Levison et al [45] reported a new approach, which combined the advantages in batch adsorption and packed bed chromatographies.…”

Section: Purification Of Hbcag From Unclarified Feedstockmentioning

confidence: 98%

Purification of recombinant hepatitis B core antigen from unclarified Escherichia coli feedstock using phage-immobilized expanded bed adsorption chromatography

Tan

Tey

2012

Journal of Chromatography B

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Isolation and purification of endo-polygalacturonase by affinity chromatography in a fluidized bed reactor

Cited by 39 publications

References 13 publications

Development of an expanded bed technique for an affinity purification of G6PDH from unclarified yeast cell homogenates

Development of an expanded bed technique for an affinity purification of G6PDH from unclarified yeast cell homogenates

Development of operating conditions for protein purification using expanded bed techniques: The effect of the degree of bed expansion on adsorption performance

Purification of recombinant hepatitis B core antigen from unclarified Escherichia coli feedstock using phage-immobilized expanded bed adsorption chromatography

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