Extractive purification of enzymes from animal tissue using aqueous two phase systems: pilot scale studies

Boland, Michael J.; Hesselink, Paul G.M.; Papamichael, N.; Hustedt, Helmut

doi:10.1016/0168-1656(91)90072-4

Cited by 33 publications

(17 citation statements)

References 7 publications

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“…The realization of aqueous two‐phase extraction (ATPE) unit operations for protein separation at industrial scales hinges not only on technical potential and feasibility, but also on economics. As discussed previously, ATPE can achieve higher yields, with cruder feeds containing more cell debris compared with processes using precipitation and conventional chromatography operations . And in general, ATPE becomes more attractive for large‐scale processes due to the higher initial yields, better clarification, savings in energy and labor, and possibility of room temperature operation , …”

Section: Introductionmentioning

confidence: 99%

“…A serious concern in the use of ATPE is the large amount of costly phase‐forming polymer needed to extract a desired protein from a complex mixture , , , . A related concern is the environmental impact and cost of disposing of large quantities of spent polymers.…”

Section: Introductionmentioning

confidence: 99%

“…They studied the effect of the first stage fresh feed/second stage fresh solvent flowrate ratio as a function of sodium chloride concentration in the feed and reported optimal steady state process conditions for the separation. There is also a more general literature base concerning the evaluation of costs associated with specific, non‐optimized ATPE‐based protein separation processes,, , , or combinations of other separation unit operations with ATPE , …”

Section: Introductionmentioning

confidence: 99%

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Towards optimal aqueous two‐phase extraction system flowsheets for protein purification

Ahmad

Przybycien

2012

J of Chemical Tech & Biotech

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BACKGROUND This work describes a flowsheet optimization framework for the development of protein purification operations involving aqueous two‐phase protein extraction (ATPE). The framework encompasses both the flowsheet and thermodynamic calculations and predicts the optimal flowsheets for the application of ATPE systems. RESULTS Results presented were generated by applying an optimization algorithm to a previous case study with demonstrated feasible solutions1 concerning the recovery of phosphofructokinase from a simple model feedstock containing both phosphofructokinase and ovalbumin using poly(ethylene glycol)‐dextran (PEG6000‐DxT500) two‐phase systems. An optimal flowsheet for a one‐stage extraction unit and improved flowsheets for two‐stage extraction units with and without recycle were determined based on the minimum scaled process cost subject to attainment of liquid‐liquid equilibrium in the ATPE system(s) and constraints on product yield, product purity and phase settling rates. The cost performance factors of feasible one‐stage and two‐stage flowsheets with and without recycle were improved by 4.7%, 8.6% and 33.1%, respectively, relative to the prior work. CONCLUSION This flowsheet optimization framework was able to predict the optimal flowsheets for protein purification from a two‐protein mixture using ATPE systems, subject to some limitations. The algorithm has the potential to be successfully applied to predict protein extraction from complex mixtures upon improvement in its computational robustness. Copyright © 2012 Society of Chemical Industry

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Towards optimal aqueous two‐phase extraction system flowsheets for protein purification

Ahmad

Przybycien

2012

J of Chemical Tech & Biotech

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“…In a formal comparison of two different technologies, technical and economical aspect needs to be considered. However, detailed economic analyses of biotechnological processes are not common [1,6,7]. In particular, the need of reports addressing the direct comparison of established and alternative processes is evident [8].…”

Section: Introductionmentioning

confidence: 99%

Direct comparison between ion-exchange chromatography and aqueous two-phase processes for the partial purification of penicillin acylase produced by E. coli

Aguilar

Albiter

Serrano‐Carreón

et al. 2006

Journal of Chromatography B

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“…It goes from purification of pharmaceutics (Harris et al, 2007) and therapeutic proteins (Asenjoet al, 2012), to isolation of proteins from bacteria (Lutwycheet al,1995) animals (Gierowet al, 1986Boland et al, 1991) and plants. In plants, the aqueous two-phase system has been mostly used for protein isolation from Arabidopsis (DeWitt et al, 1996;Borneret al, 2003).…”

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confidence: 99%

Evaluation of four different strategies to characterize plasma membrane proteins from banana roots

Garcia

Panis²,

Swennen

et al. 2014

Ciênc. agrotec.

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Plasma membrane proteins constitute a very important class of proteins. They are involved in the transmission of external signals to the interior of the cell and selective transport of water, nutrients and ions across the plasma membrane. However, the study of plasma membrane proteins is challenging because of their poor solubility in aqueous media and low relative abundance. In this work, we evaluated four different strategies for the characterization of plasma membrane proteins from banana roots: (i) the aqueous-polymer two-phase system technique (ATPS) coupled to gelelectrophoresis (gel-based), and (ii) ATPS coupled to LC-MS/MS (gel free), (iii) a microsomal fraction and (iv) a full proteome, both coupled to LC-MS/ MS. Our results show that the gel-based strategy is useful for protein visualization but has major limitations in terms of time reproducibility and efficiency. From the gel-free strategies, the microsomal-based strategy allowed the highest number of plasma membrane proteins to be identified, followed by the full proteome strategy and by the ATPS based strategy. The high yield of plasma membrane proteins provided by the microsomal fraction can be explained by the enrichment of membrane proteins in this fraction and the high throughput of the gel-free approach combined with the usage of a fast high-resolution mass spectrometer for the identification of proteins.Index terms: Aqueous-polymer two-phase system technique, microsomal and full proteome fractions, protein identification. RESUMOProteínas da membrana plasmática constituem uma importante classe de proteínas. Elas estão envolvidas na transmissão de sinais externos para o interior da célula e no transporte seletivo de nutrientes/ions, através da membrana plasmática. Porém, o estudo dessas proteinas é difícil, porque elas são poucoabundantes e apresentam baixa solubilidade em tampões aquosos. Neste trabalho, nós avaliamos quatro estratégias diferentes para extrair proteínas da membrana plasmática de raízes de banana: (i) a técnica de sistema aquoso de duas fases, constituída porpolímeros(ATPS) combinada com eletroforese em gel e (ii) ATPS sem gel usandoLC-MS/MS, (iii) uma fração microssomal e (iv) uma fração, contendo o proteoma total celular, ambas as frações avaliadas via LC-MS/MS. Nossos resultados mostram que a estratégia baseada em eletroforese em gel é útil para a visualização de proteínas, mas apresenta limitações em termos de reproducibilidade e eficiência. Dentre as estratégias sem o uso de gel, a fração microssomal permitiu a identificação do maior número de proteínas de membrana plasmática, seguida pela fração de proteoma total e pela técnica de sistema aquoso de duasfases. O alto rendimento de proteínas de membrana plasmática proporcionado pela fração microssomal pode ser explicado pelo enriquecimento de proteínas de membrana nessa fração e pela eficiência do espectrômetro de massa na identificação de proteínas.Termos para indexação: Sistema aquoso de duas fases, frações microssomal e de proteoma total, identificação de proteín...

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Extractive purification of enzymes from animal tissue using aqueous two phase systems: pilot scale studies

Cited by 33 publications

References 7 publications

Towards optimal aqueous two‐phase extraction system flowsheets for protein purification

Towards optimal aqueous two‐phase extraction system flowsheets for protein purification

Direct comparison between ion-exchange chromatography and aqueous two-phase processes for the partial purification of penicillin acylase produced by E. coli

Evaluation of four different strategies to characterize plasma membrane proteins from banana roots

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