Characterisation and separation of infectious bursal disease virus-like particles using aqueous two-phase systems

Infectious bursal disease(IBD)causes considerable economic losses in the commercial poultry industry worldwide.The principal way to control IBD virus(IBDV),the causative agent of IBD,is still through vaccination programs.Virus-like particles(VLPs)are recognised as a safe and potent recombinant vaccine platform.This research work explores the characterisation and separation of infectious bursal disease virus-like particles(IBD-VLPs)from crude feedstock.Various characteristics were studied with highperformance size-exclusion chromatography(HP-SEC),sodium dodecyl sulphate–polyacrylamide gel electrophoresis(SDS-PAGE)and transmission electron microscopy(TEM)analyses.Subsequently,the separation of IBD-VLPs using polyethylene glycol(PEG)/sodium citrate-based aqueous two-phase systems(ATPSs)was conducted and optimised.Moreover,a scale-up study of the best ATPS constituted of 15%PEG 6000,11%sodium citrate and 10%crude feedstock was performed to compare the separation performance of IBD-VLPs with and without centrifugation-assisted.The results indicated that the optimised ATPS with centrifugation-assisted for both 5 g and 50 g systems showed good recovery of IBDVLPs of>97%in the interphase between the PEG-rich top and salt-rich bottom phases.These optimised systems also showed high removal efficiencies of impurities of>95%.The results demonstrated that aqueous two-phase extraction could be a promising technology for efficient VLPs separation.

Study on aqueous two-phase systems of the mixture SDS/CTAB surfactants

The phenomenon of two dilute aqueous phases composed of sodium dodecylsulfate(SDS) and cetyl trimethyl ammonium bromide(CTAB) was investigated under various conditions such as concentrations and molar ratios of the two surfactants, the addition of sodium chloride and temperature. Vesicles formation was found in the both phases by TEM image.

Separation/Analysis Rhodamine B by Anion Surfactant/Ionic Liquid Aqueous Two-Phase Systems Coupled with Ultraviolet Spectrometry

A novel method for the separation/analysis rhodamine B has been described. The ionic liquid (1-tetradecyl-3-methylimidazolium bromide)/anion surfactant (sodium dodecyl sulfate)/NaCl two-phase systems (ATPSs) is presented as a simple, rapid and effective sample pretreatment technique coupled with ultraviolet spectrometry for analysis rhodamine B in soft drink. The effects of parameters on the ATPSs extraction of rhodamine B such as amount of surfactant, ionic liquid and salt, pH, temperature, stabilization and centrifugal time have been studied in details. Under the optimized conditions, the linear range of calibration curve for rhodamine B was 0.05 - 7.0 μg·mL-1 and the detection limit was 3.2 ng·mL-1. The phase equilibrium and the mechanism of phase separation for ATPSs have been discussed. This method has been applied to the determination of rhodamine B in soft drink.

Aqueous Two-Phase Systems Applied to Partition Proteins from Goat Milk Whey In-Nature

The proteins coming from the milk whey have numerous functional properties. Among the proteins with high bioactivity, α-lactoalbumin (α-La) and β-lactoglobulin (β-Lg) are present in large quantities in the milk whey. In the separation process of proteins, it is important to choose techniques which besides ensuring purity and high yield will not affect the molecule biological activity. The aqueous two-phase systems (ATS) have been utilized with success in the partition of these proteins, however, the studies were performed using protein in its pure form. Studies using milk whey in-nature and goat milk whey have not been found yet. In this context, the objective of this study was to evaluate the liquid liquid equilibrium of aqueous two-phase systems (ATS) in the partition of α-La and β-Lg from goat milk whey in-nature. Equilibrium data were performed considering ATS comprised of polyethylene glycol, potassium phosphate and water at 25°C and pH 7.0. The influence of the polymer molecular weight and amount of goat milk whey in-nature on the partition coefficient of these proteins were assessed. The partition coefficient, selectivity, process yield and purity of α-lactoalbumin and β-lactoglobulin proteins were determined. The results showed that the separation technique by aqueous biphasic systems is applicable indicating high efficiency in the whey proteins separation process.

Emerging delivery systems based on aqueous two-phase systems:A review

The aqueous two-phase system(ATPS)is an all-aqueous system fabricated from two immiscible aqueous phases.It is spontaneously assembled through physical liquid-liquid phase separation(LLPS)and can create suitable templates like the multicompartment of the intracellular environment.Delicate structures containing multiple compartments make it possible to endow materials with advanced functions.Due to the properties of ATPSs,ATPS-based drug delivery systems exhibit excellent biocompatibility,extraordinary loading efficiency,and intelligently controlled content release,which are particularly advantageous for delivering drugs in vivo.Therefore,we will systematically review and evaluate ATPSs as an ideal drug delivery system.Based on the basic mechanisms and influencing factors in forming ATPSs,the transformation of ATPSs into valuable biomaterials is described.Afterward,we concentrate on the most recent cutting-edge research on ATPS-based delivery systems.Finally,the potential for further collaborations between ATPS-based drug-carrying biomaterials and disease diagnosis and treatment is also explored.

Aqueous Two-phase Systems with Ultrasonic Extraction Used for Extracting Phenolic Compounds from Inonotus obliquus

Objective To optimize the extracting technology of assessing the maximum yield of phenolic compounds (PC) from Inonotus obliquus by single factor experiments and orthogonal array design methods through aqueous two-phase systems combined with ultrasonic extraction. Methods The range of the independent variables, namely levels of acetone and ammonium sulfate, and ultrasonic time were identified by a first set of single factor experiments. The actual values of the independent variables coded at four levels and three factors were selected based on the results of the single factor experiments. Subsequently, the levels of acetone and ammonium sulfate, and ultrasonic time were optimized using the orthogonal array method. Results The optimum conditions for the extraction of PC were found to use 7.0 mL acetone, 5.5 mg ammonium sulfate, with ultrasonic time for 5 min. Under these optimized conditions, the experimental maximum yield of PC was 37.8 mg/g, much higher than that of the traditional ultrasonic extraction (UE, 29.0 mg/g). And the PC obtained by this method had stronger anti-oxidative activities than those by traditional UE method. Conclusion These results indicate the suitability of the models developed and the success in optimizing the extraction conditions. This is an economical and efficient method for extracting polyphenols from I. obliquus.

Selective separation of protein and saccharides by ionic liquids aqueous two-phase systems

In the present work,it was found that aqueous solution of a hydrophilic ionic liquid (IL),1-butyl-3-methylimidazolium dicyanamide ([C4mim][N(CN)2]),could be separated into an aqueous two-phase system (ATPS) by inorganic salts such as K2HPO4 and K3PO4.The top phase is IL-rich,while the bottom phase is phosphate-rich.It was shown that 82.7%-100% bovine serum albumin (BSA) could be enriched into the top phase and almost quantitative saccharides (arabinose,glucose,sucrose,raffinose or dextran) were preferentially extracted into the bottom phase in a single-step extraction by [C4mim][N(CN)2] + K2HPO4 ATPS.The extraction efficiency of BSA from the aqueous saccharide solutions was influenced by the molecular structure of saccharides.The conductivity,dynamic light scattering (DLS) and transmission electron microscopy (TEM) were combined to investigate the microstructure of the IL-rich top phase and the possible mechanism for the selective separation.It is suggested that the formation of the IL aggregate and the IL aggregate-BSA complex plays a significant role in the separation of BSA from aqueous saccharide solutions.This is the first example for the selective separation by ILs-based ATPSs.It is expected that these findings would have potential applications in bio-analysis,separation,and IL recycle.

Recovery of natural active molecules using aqueous two-phase systems comprising of ionic liquids/deep eutectic solvents

Due to environmental protection requirements,extraction of bioactive compounds from plant materials using environment-friendly green solvents has always been a research hotspot.And great efforts of scholars have been made in this direction,as well as environment-friendly solvents have been used to develop many innovative extraction techniques.Ionic liquids(ILs)and deep eutectic solvents(DESs)are two kinds of typical designable green solvents,which are potential replacements for traditional volatile organic solvents used for extracting.Under the substances action of inorganic salts or polymers,ILs/DESs can form an aqueous two-phase system(ATPS),which has obvious advantages for separating natural products.This paper discussed the phase separation principle of ILs/DESs-based ATPSs and reviewed the applications in the extraction of natural active molecules in recent years,as well as to promote the development of separation of the active constituents in Chinese materia medica.

Partitioning of biomaterials in aqueous two-phase systems of mixed cationic-anionic surfactants

In a previous paper, we have showed that, when aqueous solutions of cationicand anionic surfactants at certain concentrations were mixed, the solution separatedspontaneously into two immiscible phases (aqueous two-phases), one phase was rich,and the other was poor in the mixed surfactants. A clear interfacial boundary existsbetween two phases.

Phase Behavior of Ionic Liquids-Cesium Carbonate-Water Aqueous Two-phase Systems and Their Extraction of L-Tryptophan

Phase behavior and extraction ability of aqueous two-phase systems（ATPs） consisting of ionic liquids（ILs）, Cs2CO3 and water were investigated in this paper. Four kinds of ionic liquids, namely, 1-amyl-3-methylimidazolium bromide（[Csmim]Br）, 1-hexyl-3-methylimidazolium bromide（[C6nim]Br）, 1-heptyl-3-methylimidazolium bromide （[C7mim]Br） and 1-octyl-3-methylimidazolium bromide（[C8mim]Br）, were examined to discuss the influence of alkyl groups. Binodal curves and tie-lines at 288.15, 298.15 and 308.15 K were obtained. The partitioning behavior for L-tryptophan in such ATPs was further investigated. The effect of temperature, pH, Cs2CO3 concentration and the structure of ionic liquids on the partitioning were discussed in detail.

Response Surface Optimization of Ultrasound-assisted Aqueous Two-phase Extraction of Sweet Potato Leaf Polysaccharides

[Objectives]The ultrasound-assisted aqueous two-phase extraction of sweet potato leaf polysaccharides was studied.[Methods]With the yield of sweet potato leaf polysaccharides as the index,the aqueous two-phase extraction system was determined,and the optimal extraction conditions were optimized by single-factor experiments and response surface methodology.[Results]The optimal parameters were ethanol concentration 25.68%,liquid-to-material ratio 55.83,and ultrasonic treatment time 38.33 min.Under these conditions,the yield of sweet potato leaf polysaccharides could reach 20.646 mg/g.[Conclusions]The ethanol/ammonium sulfate aqueous system is a rapid and efficient method for extracting sweet potato leaf polysaccharides,which is of great significance for the application of sweet potato leaf extract as a natural food additive.

Measurement and Correlation of Equilibrium Data for Aqueous Two-phase System Ethanol+Water+ K_2HPO_4

The isothermal solubility data of aqueous two-phase system ethanol+water+K 2HPO 4 were determined with the turbidity titration method at 303.2 K. The binodal curves were described by using the Mistry equation very well. An experimental procedure for measuring the liquid-liquid equilibrium data of the aqueous two-phase system was proposed, in which the concentrations of the coexisting phases were determined with the corresponding densities of the solution. The tie lines were satisfactorily described by using the Othmer-Tobias and Bancroft equations.

EXTRACTION OF INTERFERON-γ USING PEG/K_3PO_4 AQUEOUS TWO-PHASE SYSTEM

The aqueous two-phase system of PEG-4000/K3PO4 is selected to extract interferon-γ（γ-IFN）from the broth of homogenized E.coil cells,which were recombined genetically and werefermented in high expressivity.The effects of pH value and the concentrations of PEG and phos-phate on the partition behavious of γ-IFN and contaminating proteins have been investigated.Theresults show that the pH value is the most influential factor effecting the extraction yield of γ-IFN.Under a suitable condition,an effective separation with high yield of γ-IFN can be achieved.

The Effect of pH Difference Between Two Phases on the Partition of Lysozyme in Aqueous Two-Phase System

In the investigation of effect of KSCN on the partitioning of lysozyme in PEG2000/ammonium sulfate aqueous two-phase system, it was found that the KSCN could alter the pH difference between the two phases. and thus affect the partition of lysozyme. The relationship between partition coefficients of lysozyme and pH differences between two phases was discussed.

Copper(Ⅰ)-catalyzed Synthesis of Aryl, Heteroaryl and Vinyl Sulfides in Aqueous Two-phase System

A fast and convenient C―S bond formation reaction was achieved catalyzed by CuI/L-proline in aqueous two-phase system providing a simple method for synthesis of aryl sulfides in good yields in short time.

Selective separation and enrichment of proteins in aqueous two-phase extraction system

A simple aqueous two-phase extraction system （ATPS） of PEG/phosphate was proposed for selective separation and enrichment of proteins. The combination of ATPE with HPLC was applied to identify the partition of proteins in two phases. Five proteins （bovine serum albumin, Cytochrome C, lysozyme, myoglobin, and trypsin） were used as model proteins to study the effect of phosphate concentration and pH on proteins partition. The PEG/phosphate system was firstly applied to real human saliva and plasma samples, some proteins showed obviously different partition in two phases. The primary results manifest the selective separation and enrichment of proteins in ATPS provided the potential for high abundance proteins depletion in proteomics. ~ 2009 Feng Qu. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Inversion of two-phase extensional basin systems during subduction of the Paleo-Pacific Plate in the SW Korean Peninsula:Implication for the Mesozoic “Laramide-style” orogeny along East Asian continental margin

During subduction, continental margins experience shortening along with inversion of extensional sedimentary basins. Here we explore a tectonic scenario for the inversion of two-phase extensional basin systems, where the Early-Middle Jurassic intra-arc volcano-sedimentary Oseosan Volcanic Complex was developed on top of the Late Triassic-Early Jurassic post-collisional sequences, namely the Chungnam Basin. The basin shortening was accommodated mostly by contractional faults and related folds. In the basement, regional high-angle reverse faults as well as low-angle thrusts accommodate the overall shortening, and are compatible with those preserved in the cover. This suggests that their spatial and temporal development is strongly dependent on the initial basin geometry and inherited structures.Changes in transport direction observed along the basement-sedimentary cover interface is a characteristic structural feature, reflecting sequential kinematic evolution during basin inversion. Propagation of basement faults also enhanced shortening of the overlying sedimentary cover sequences. We constrain timing of the Late Jurassic-Early Cretaceous(ca. 158-110 Ma) inversion from altered K-feldspar 40 Ar/39 Ar ages in stacked thrust sheets and K-Ar illite ages of fault gouges, along with previously reported geochronological data from the area. This "non-magmatic phase" of the Daebo Orogeny is contemporaneous with the timing of magmatic quiescence across the Korean Peninsula. We propose the role of flat/low-angle subduction of the Paleo-Pacific Plate for the development of the "Laramide-style" basement-involved orogenic event along East Asian continental margin.

Aqueous Two-Phase Interfacial Assembly of COF Membranes for Water Desalination

Aqueous two-phase system features with ultralow interfacial tension and thick interfacial region,affording unique confined space for membrane assembly.Here,for the first time,an aqueous two-phase interfacial assembly method is proposed to fabricate covalent organic framework(COF)membranes.The aqueous solution containing polyethylene glycol and dextran undergoes segregated phase separation into two water-rich phases.By respectively distributing aldehyde and amine monomers into two aqueous phases,a series of COF membranes are fabricated at water-water interface.The resultant membranes exhibit high NaCl rejection of 93.0-93.6% and water permeance reaching 1.7-3.7 L m^(−2) h^(−1) bar^(−1),superior to most water desalination membranes.Interestingly,the interfacial tension is found to have pronounced effect on membrane structures.The appropriate interfacial tension range(0.1-1.0 mN m^(−1))leads to the tight and intact COF membranes.Furthermore,the method is extended to the fabrication of other COF and metal-organic polymer membranes.This work is the first exploitation of fabricating membranes in all-aqueous system,confering a green and generic method for advanced membrane manufacturing.

Hydrolysis hydrogen production mechanism of Mg10Ni10Ce alloy surface modified by SnO_(2) nanotubes in different aqueous systems

(Mg-10wt%Ni)-10wt%Ce(Mg10Ni10Ce)was ball-milled with SnO_(2)nanotubes and Mg10Ni10Ce-xSnO_(2)(x¼0,5,10 and 15 wt%)composites have been prepared.The phase compositions,microstructures,morphologies and hydrolysis H2 generation performance in different aqueous systems(distilled water,tap water and simulated seawater)have been investigated and the corresponding hydrolysis mechanism of Mg10Ni10Ce and Mg10Ni10CeeSnO_(2)has been proposed.Adding a small amount of SnO_(2)nanotubes can significantly enhance the hydrolysis reaction of Mg10Ni10Ce,especially the initial hydrolysis kinetics and the final H_(2) generation yield.Unfortunately,the Mg10Ni10Ce-xSnO_(2)hardly reacts with distilled water at room temperature.The hydrolysis reaction rate of Mg10Ni10Cee5SnO_(2)composite in tap water is still very slow with only 17.3%generation yield after 1 h at 303 K.Fortunately,in simulated seawater(3.5 wt%NaCl solution),the hydrolytic H2 generation behavior of the Mg10Ni10Cee5SnO_(2)composite has been greatly improved,which can release as high as 468.6 mL g^(-1 )H_(2) with about 60.9%generation yield within 30 s at 303 K.The Cl destroys the passivation layer on MgeNieCe alloy surface and the added SnO_(2)nanotubes accelerate the hydrolysis reaction rate and enhance the H2 generation yield.The Mg10Ni10Cee5SnO_(2)composite can rapidly generate a large amount of H2 in simulated seawater in a short time,which is expected to be applied on portable H2 generators in the future.