Purification of Alkaline Phosphatase by Ultrafiltration in a Stirred Batch Cell

Slater, C. Stewart; Huggins, T. G.; Brooks, Clayton A.; Hollein, Helen C.

doi:10.1080/01496398608056136

Cited by 5 publications

(2 citation statements)

References 11 publications

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“…Ultrafiltration (UF), a pressure-driven membrane-based separation process, with an inherent high throughput and ever-improving selectivity of separation, has emerged as a powerful bioseparation technique for large-scale fractionation and polishing of protein products such as therapeutic drugs, enzymes, vaccines, and antibodies. Indeed, extensive literature has demonstrated that macromolecular species having similar molecular weights can be satisfactorily separated through the fine-tuning of system properties and operational parameters to manipulate protein−protein and protein−membrane interactions and to control concentration polarization and membrane fouling. − …”

Section: Introductionmentioning

confidence: 99%

“…Indeed, extensive literature has demonstrated that macromolecular species having similar molecular weights can be satisfactorily separated through the fine-tuning of system properties and operational parameters to manipulate protein-protein and protein-membrane interactions and to control concentration polarization and membrane fouling. [10][11][12][13][14][15][16][17][18][19][20][21][22][23][24] A number of studies on the separation of lysozyme using UF have been reported. Balakrishnan and Agarwal 14 investigated the fractionation of simulated mixtures of lysozyme/ovalbumin and lysozyme/myoglobin in a vortex flow ultrafiltration.…”

Section: Introductionmentioning

confidence: 99%

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Fractionation of Lysozyme and Chicken Egg Albumin Using Ultrafiltration with 30-kDa Commercial Membranes

Wang

Cui

2005

Ind. Eng. Chem. Res.

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Lysozyme is an important protein in chicken egg white (CEW) and it is feasible to separate lysozyme from other CEW proteins using ultrafiltration. To identify suitable conditions for the fractionation of lysozyme from CEW, we studied with the model binary mixture of lysozyme and chicken egg albumin (CEA) with the composition and concentration of the two proteins similar to those in natural CEW. Two commercial ultrafiltration (UF) membranes of 30-kDa molecular weight cutoff (MWCO) made of regenerated cellulose (RC) and poly(ether sulfone) (PES) membranes were used. By systematically examining the effect of the physicochemical properties of the solution and the hydrodynamics of the membrane systems, the most suitable conditions to achieve maximum transmission and selectivity for lysozyme were identified. The PES UF membranes offer distinct advantages for the fractionation of lysozyme from this binary mixture. With the 30-kDa PES UF membrane, the suitable conditions were identified as pH 10-11, salt concentration (NaCl) 100 mM, permeate flux 26.5 L/m 2 h, and stirring speed 2100 rpm. Under these conditions, a lysozyme transmission of 99% was obtained and the selectivity of separation was as high as 2400.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fractionation of Lysozyme and Chicken Egg Albumin Using Ultrafiltration with 30-kDa Commercial Membranes

Wang

Cui

2005

Ind. Eng. Chem. Res.

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Fractionation of Proteins Using Ultrafiltration: Developments and Challenges

Wan

Cui

Ghosh

2005

Dev. Chem. Eng. Mineral Process.

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I In recent years ultrafiltration has attracted significant interest as a potential technology for fractionating proteins. Although traditionally thought to be suitable for size-based separation with high-throughput but relatively low-resolution, recent research has demonstrated that it is possible to signijicantly increase the selectivity in protein fractionation using ultrafiltration while still maintaining its inherent highthroughput. This paper reviews recent developments in the area of protein pactionation using ultrafiltration, with focus on strategies employed to improve selectivity of separation. Several new techniques, technological developments, and their applications for selective ultrajiltration of proteins are also briejly introduced. IntroductionWith the rapid growth in the field of biotechnology, the need for reducing production costs coupled with projected increases in batch size requires development of protein purification processes with high-throughput as well as hgh-resolution. Ultrafiltration has the potential to meet both these challenges. It is very well suited to the processing of biological products since it operates at relatively "mild" conditions and involves no phase change or addition of chemicals. Thls minimizes the extent of denaturation, deactivation, andor degradation of the highly labile biological molecules. A significant advantage of ultrafiltration is the high throughput of product, whch is attractive in biotechnological processes. Moreover, it is much less expensive than other separation and purification methods, e.g. chromatography, and is easy to operate and to scale up. All of these make it very promising for biotechnological applications.* Author for correspondence ('hanfeng.cui@eng.ox.ac.uk). I 2 I Y. Wan, R. Ghosh andZ. CuiThe most common applications of ultrafiltration in downstream processing are protein concentration (i.e. solvent removal), buffer exchange and desalting, virus removal and clarification. In recent years, there has been growing interest in the use of ultrafiltration for fractionating complex protein mixtures, commonly encountered in many biotechnological, food and biomedical applications. However, protein fractionation using ultrafiltration remains a technical challenge, and ultrafiltrationbased processes have not been commercialised to any significant extent. In most cases, for the proteins to be separated having similar sizes and consequently obtaining sufficient selectivity using available membranes is not a trivial matter.Early attempts to use membrane technology for fractionation of protein mixtures were often unsuccessful due to the fairly limited selectivity offered by the membrane systems at the chosen operating conditions. This led to the myth that effective protein separation could only be obtained for proteins differing by at least a factor of ten in terms of molecular weight, a rule of thumb that is still used quite extensively in the initial phases of development of ultrafiltration-based protein separation processes [ 11. Nakatsuka and Mich...

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Protein fractionation in a vortex flow filter. II: Separation of simulated mixtures

Balakrishnan

Agarwal

1996

Journal of Membrane Science

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Purification of Alkaline Phosphatase by Ultrafiltration in a Stirred Batch Cell

Cited by 5 publications

References 11 publications

Fractionation of Lysozyme and Chicken Egg Albumin Using Ultrafiltration with 30-kDa Commercial Membranes

Fractionation of Lysozyme and Chicken Egg Albumin Using Ultrafiltration with 30-kDa Commercial Membranes

Fractionation of Proteins Using Ultrafiltration: Developments and Challenges

Protein fractionation in a vortex flow filter. II: Separation of simulated mixtures

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