Source:http://linkedlifedata.com/resource/pubmed/id/10578095
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
4
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| pubmed:dateCreated |
2000-2-7
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| pubmed:abstractText |
In this study we show that proteins can be partitioned and separated in a novel aqueous two-phase system composed of only one polymer in water solution. This system represents an attractive alternative to traditional two-phase systems which uses either two polymers (e.g., PEG/dextran) or one polymer in high-salt concentration (e.g., PEG/salt). The polymer in the new system is a linear random copolymer composed of ethylene oxide and propylene oxide groups which has been hydrophobically modified with myristyl groups (C(14)H(29)) at both ends (HM-EOPO). This polymer thermoseparates in water, with a cloud point at 14 degrees C. The HM-EOPO polymer forms an aqueous two-phase system with a top phase composed of almost 100% water and a bottom phase composed of 5-9% HM-EOPO in water when separated at 17-30 degrees C. The copolymer is self-associating and forms micellar-like structures with a CMC at 12 microM (0.01%). The partitioning behavior of three proteins (lysozyme, bovine serum albumin, and apolipoprotein A-1) in water/HM-EOPO two-phase systems has been studied, as well as the effect of various ions, pH, and temperature on protein partitioning. The amphiphilic protein apolipoprotein A-1 was strongly partitioned to the HM-EOPO-rich phase within a broad-temperature range. The partitioning of hydrophobic proteins can be directed with addition of salt. Below the isoelectric point (pI) BSA was partitioned to the HM-EOPO-rich phase and above the pI to the water phase when NaClO(4)was added to the system. Lysozyme was directed to the HM-EOPO phase with NaClO(4), and to the water phase with Na-phosphate. The possibility to direct protein partitioning between water and copolymer phases shows that this system can be used for protein separations. This was tested on purification of apolipoprotein A-1 from human plasma and Escherichia coli extract. Apolipoprotein A-1 could be recovered in the HM-EOPO-rich phase and the majority of contaminating proteins in the water phase. By adding a new water/buffer phase at higher pH and with 100 mM NaClO(4), and raising the temperature for separation, the apolipoprotein A-1 could be back-extracted from the HM-EOPO phase into the new water phase. This novel system has a strong potential for use in biotechnical extractions as it uses only one polymer and can be operated at moderate temperatures and salt concentrations and furthermore, the copolymer can be recovered.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Apolipoprotein A-I,
http://linkedlifedata.com/resource/pubmed/chemical/Ions,
http://linkedlifedata.com/resource/pubmed/chemical/Micelles,
http://linkedlifedata.com/resource/pubmed/chemical/Muramidase,
http://linkedlifedata.com/resource/pubmed/chemical/Polyethylenes,
http://linkedlifedata.com/resource/pubmed/chemical/Polymers,
http://linkedlifedata.com/resource/pubmed/chemical/Polypropylenes,
http://linkedlifedata.com/resource/pubmed/chemical/Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Salts,
http://linkedlifedata.com/resource/pubmed/chemical/Serum Albumin, Bovine,
http://linkedlifedata.com/resource/pubmed/chemical/UCON 50-HB-5100,
http://linkedlifedata.com/resource/pubmed/chemical/Water
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| pubmed:status |
MEDLINE
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| pubmed:issn |
0006-3592
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| pubmed:author | |
| pubmed:copyrightInfo |
Copyright 1999 John Wiley & Sons, Inc.
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| pubmed:issnType |
Print
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| pubmed:volume |
66
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
247-57
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| pubmed:dateRevised |
2006-11-15
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| pubmed:meshHeading |
pubmed-meshheading:10578095-Animals,
pubmed-meshheading:10578095-Apolipoprotein A-I,
pubmed-meshheading:10578095-Biotechnology,
pubmed-meshheading:10578095-Cattle,
pubmed-meshheading:10578095-Escherichia coli,
pubmed-meshheading:10578095-Humans,
pubmed-meshheading:10578095-Hydrogen-Ion Concentration,
pubmed-meshheading:10578095-Ions,
pubmed-meshheading:10578095-Micelles,
pubmed-meshheading:10578095-Muramidase,
pubmed-meshheading:10578095-Polyethylenes,
pubmed-meshheading:10578095-Polymers,
pubmed-meshheading:10578095-Polypropylenes,
pubmed-meshheading:10578095-Proteins,
pubmed-meshheading:10578095-Recombinant Proteins,
pubmed-meshheading:10578095-Salts,
pubmed-meshheading:10578095-Serum Albumin, Bovine,
pubmed-meshheading:10578095-Temperature,
pubmed-meshheading:10578095-Water
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| pubmed:year |
1999
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| pubmed:articleTitle |
Thermoseparating water/polymer system: a novel one-polymer aqueous two-phase system for protein purification.
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| pubmed:affiliation |
Department of Biochemistry, Lund University, P.O.B. 124, S-221 00, Lund, Sweden.
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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