Thermal-aggregation suppression of proteins by a structured PEG analogue: Importance of denaturation temperature for effective aggregation suppression

Muraoka, Takahiro; Sadhukhan, Nabanita; Ui, Mihoko; Kawasaki, Shunichi; Hazemi, Enrikko; Adachi, Kota

doi:10.1016/j.bej.2014.03.001

Cited by 11 publications

(3 citation statements)

References 23 publications

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“…Thus, and although polymers such as PEG are used in the precipitation and aggregation of proteins [43], the amount of water in the PEG-rich phase, as well as its low molecular weight, is favorable for maintaining a protein-friendly environment for ovalbumin. Previously, it was showed that chicken egg white lysozyme tends to cluster into aggregates in presence of PEG 1000 after incubation at 90 °C for 30 min [44][45]. At high temperatures, and due to the lower critical solution temperature behavior of PEG-water systems, there is a decrease on the hydration shell of the proteins [44][45].…”

Section: Extraction/separation Of Ovalbumin Directly From Egg Whitementioning

confidence: 99%

“…Previously, it was showed that chicken egg white lysozyme tends to cluster into aggregates in presence of PEG 1000 after incubation at 90 °C for 30 min [44][45]. At high temperatures, and due to the lower critical solution temperature behavior of PEG-water systems, there is a decrease on the hydration shell of the proteins [44][45]. In this work, the extractions were carried out at a low temperature, 25ºC, and using polymers of low molecular weight (from 400 to 1000 mol·kg -1 ).…”

Section: Extraction/separation Of Ovalbumin Directly From Egg Whitementioning

confidence: 99%

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Single-step purification of ovalbumin from egg white using aqueous biphasic systems

Pereira

Cruz

Almeida

et al. 2016

Process Biochemistry

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The ability of aqueous biphasic systems (ABS) composed of polyethylene glycols of different molecular weights (PEG 400, 600 and 1000) and buffered aqueous solutions of potassium citrate/ citric acid (pH = 5.0 -8.0) to selectively extract ovalbumin from egg white was here investigated. Phase diagrams, tie-lines and tie-line lengths were determined at 25ºC and the partitioning of ovalbumin in these systems was then evaluated. Aiming at optimizing the selective extraction of ovalbumin in the studied ABS, factors such as pH, PEG molecular weight and amount of the phase-forming components were initially investigated with pure commercial ovalbumin. In almost all ABS, it was observed a preferential partitioning of ovalbumin to the polymer-rich phase, with extraction efficiencies higher than 90%. The best ABS were then applied in the purification of ovalbumin from the real egg white matrix. In order to ascertain on the ovalbumin purity and yield, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion high performance liquid chromatography (SE-HPLC) analyses were conducted, confirming that the isolation/purification of ovalbumin from egg white was completely achieved in a single-step with a recovery yield of 65%. The results obtained show that polymer-salt-based ABS allow the selective extraction of ovalbumin from egg white with a simpler approach and better performance than previously reported. Finally, it is shown that ovalbumin can be completely recovered from the PEG-rich phase by an induced precipitation using an inexpensive and sustainable separation platform which can be easily applied on an industrial scale.

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Section: Extraction/separation Of Ovalbumin Directly From Egg Whitementioning

confidence: 99%

Section: Extraction/separation Of Ovalbumin Directly From Egg Whitementioning

confidence: 99%

Single-step purification of ovalbumin from egg white using aqueous biphasic systems

Pereira

Cruz

Almeida

et al. 2016

Process Biochemistry

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“…Proteins generally require a formulation excipient(s) as a protein stabilizer in a liquid state. Protein stabilization by a stabilizer(s) can be achieved through the traditional preferential interaction mechanism and/or other proposed mechanisms such as nonspecific interaction with surface hydrophobic pockets or charged amino acids, specific ligand binding, and enhancement of solution viscosity . To enhance the stability of proteins, simultaneous use of multiple stabilizers has been tested in expectation of addressing different stability issues via different mechanisms and/or possible synergistic effect.…”

Section: Use Of Complex or Uncommon Stabilizersmentioning

confidence: 99%

Advanced protein formulations

Wang

2015

Protein Science

100

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It is well recognized that protein product development is far more challenging than that for small-molecule drugs. The major challenges include inherent sensitivity to different types of stresses during the drug product manufacturing process, high rate of physical and chemical degradation during long-term storage, and enhanced aggregation and/or viscosity at high protein concentrations. In the past decade, many novel formulation concepts and technologies have been or are being developed to address these product development challenges for proteins. These concepts and technologies include use of uncommon/combination of formulation stabilizers, conjugation or fusion with potential stabilizers, site-specific mutagenesis, and preparation of nontraditional types of dosage forms-semiaqueous solutions, nonfreeze-dried solid formulations, suspensions, and other emerging concepts. No one technology appears to be mature, ideal, and/or adequate to address all the challenges. These gaps will likely remain in the foreseeable future and need significant efforts for ultimate resolution.

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Effect of Photoinduced Size Changes on Protein Refolding and Transport Abilities of Soft Nanotubes

Kameta

Akiyama

Masuda

et al. 2016

Chemistry A European J

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Self-assembly of azobenzene-modified amphiphiles (Glyn Azo, n=1-3) in water at room temperature in the presence of a protein produced nanotubes with the protein encapsulated in the channels. The Gly2 Azo nanotubes (7 nm internal diameter [i.d.]) promoted refolding of some encapsulated proteins, whereas the Gly3 Azo nanotubes (13 nm i.d.) promoted protein aggregation. Although the 20 nm i.d. channels of the Gly1 Azo nanotubes were too large to influence the encapsulated proteins, narrowing of the i.d. to 1 nm by trans-to-cis photoisomerization of the azobenzene units of the Gly1 Azo monomers packed in the solid bilayer membranes led to a squeezing out of the proteins into the bulk solution and simultaneously enhanced their refolding ratios. In contrast, photoinduced transformation of the Gly2 Azo nanotubes to short nanorings (<40 nm) with a large i.d. (28 nm) provided no further refolding assistance. We thus demonstrate that pertubation by the solid bilayer membrane wall of the nanotubes is important to accelerate refolding of the denatured proteins during their transport in the narrow nanotube channels.

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Thermal-aggregation suppression of proteins by a structured PEG analogue: Importance of denaturation temperature for effective aggregation suppression

Cited by 11 publications

References 23 publications

Single-step purification of ovalbumin from egg white using aqueous biphasic systems

Single-step purification of ovalbumin from egg white using aqueous biphasic systems

Advanced protein formulations

Effect of Photoinduced Size Changes on Protein Refolding and Transport Abilities of Soft Nanotubes

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