Aqueous two-phase systems for biomolecule separation

Diamond, Alan D.; Hsu, Jong-Ping

doi:10.1007/bfb0046198

Cited by 105 publications

(92 citation statements)

References 157 publications

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“…The ligand is coupled by covalent bonds to the polymer phase; therefore it will have a high partition coefficient towards the phase enriched with the polymer. If the target protein to be isolated has a specific affinity for the ligand, the formation of a ligand-biomolecule complex will induce an increase in the partition coefficient of the protein towards the phase enriched in the ligand (5,15), while the presence of contaminant molecules will be predominant in the opposite phase (9).…”

Section: Partitioning In the Atpsmentioning

confidence: 99%

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Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning

Silva

Franco

1999

Rev. Microbiol.

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This work investigated the partitioning of β-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-β-Dgalactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme β-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where β-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of β-galactosidase (K = 2 x 10 -5 ) in order to provide the purification and recovery of 39% of the enzyme in the bottom salt-rich phase.

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Section: Partitioning In the Atpsmentioning

confidence: 99%

“…An ATPS contains a large amount of water in both phases, constituting an excellent mild biological method to recover cells, organelles or active proteins (2). Methods described in the literature, such as liquid-liquid extraction, are gaining prominence in meeting the basic demand due to the viability of their industrial applications (9).…”

Section: Introductionmentioning

confidence: 99%

Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning

Silva

Franco

1999

Rev. Microbiol.

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“…Usually, biological material has been purified by precipitation with salts or organic solvents, and by using various chromatographic techniques, all of them having enormous difficulties in large-scale applications (Alves et al, 2000). Besides, according to Diamond and Hsu (1992), 50-90% of the biological products production costs are determined by the purification strategy.…”

Section: Introductionmentioning

confidence: 99%

Large scale purification of Clostridium perfringens toxins: a review

Cavalcanti¹,

Porto²,

Porto³

et al. 2004

Rev. Bras. Cienc. Farm.

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Clostridium perfringens, a Gram-positive anaerobic bacterium, is widespread in the environment and commonly found in the intestines of animals, including humans. C. perfringens strains are classified into five toxinotypes (A, B, C, D and E) based on the production of four major toxins (α, β, ε, ι). However the toxins (theta, delta, lambda and enterotoxin) are also synthesized by C. perfringens strain. Many attempts to purify the toxins produced by C. perfringens have been proposed. In this review we discuss the purification methods used to isolate toxins from C. perfringens reported in last four decades.

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“…Furthermore, ATPS can remove contaminants such as nucleic acids and undesirable proteins. Hence, ATPS has been recognized as an efficient and economical downstream processing method due to the ease and lower cost [22,28].…”

mentioning

confidence: 99%

Partitioning and recovery of proteinase from tuna spleen by aqueous two-phase systems

Klomklao

Benjakul

Visessanguan

et al. 2005

Process Biochemistry

102

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Partitioning of spleen proteinase from yellowfin tuna (Thunnus albacores) in an aqueous two-phase system (ATPS) was investigated. Phase compositions including PEG molecular mass and concentration as well as types and concentration of salts affected the protein partitioning. ATPS comprising PEG1000 (15% w/w) and magnesium sulfate (20% w/w) provided the best condition for the maximum partitioning of the proteinase into the top phase and gave a highest specific activity (47.0 units/µg protein) and purification fold (6.61). The yield of 69.0% was obtained.Under the same ATPS condition used, the partitioning of proteinase of splenic extract from three tuna species involving skipjack tuna, yellowfin tuna and tongol tuna were compared. The purity of splenic extract from all tuna species increased after ATPS process. Among all species tested, yellowfin tuna showed the highest purification fold, followed by tongol tuna and skipjack tuna, respectively. SDS-substrate gel electrophoresis revealed that the band intensity of major proteinase in ATPS fraction from all tuna species slightly increased with the concomitant decrease in band intensity of other contaminating proteins, indicating the greater specific activity of splenic extract. Therefore, ATPS was an effective method for partitioning and recovery of proteinases from tuna spleen.

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Aqueous two-phase systems for biomolecule separation

Cited by 105 publications

References 157 publications

Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning

Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning

Large scale purification of Clostridium perfringens toxins: a review

Partitioning and recovery of proteinase from tuna spleen by aqueous two-phase systems

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