High Performance Hydrophobic Interaction Chromatography-A New Approach to Separate Intermediates of Protein Folding──Ⅰ.Separation of Intermediates of Urea-unfolded α-Amylase

Based on the different hydrophobicities of the intermediates of proteins the various conformational intermediates of the refolding of a-amylase originally denatured with 8.0 mol/L urea solution were separated with high performance hydrophobic interaction chromatography(HPHIC). Compared to the separation of the same intermediates with weak anion exchange chromatography and size-exclusion chromatography the result obtained with HPHIC is the best It would be expected that HPHIC may be a strongly potential tool to separate intermediates of some proteins which cannot be, or cannot completely be refolded by HPHIC.

Comparison of the Contributions of Tetrahydrofurfuryl Alcohol and PEG to a-Chymotrypsin Renaturation with High Performance Hydrophobic Interaction Chromatography

The contributions of tetrahydrofurfuryl alcohol (THFA) and polyethylene glycol (PEG) to the renatured efficiency of a-chymotrypsin were investigated and compared with each other. The maximum increments of bioactivity recovery of a-Chy were found to be 25.1% for THFA, 10.4% for PEG, respectively. The experimental results indicated that the denaturant solution containing THFA contributed more to the renaturation of a-Chy in high performance hydrophobic interaction chromatography (HPHIC) than that containing PEG, when the concentration of THFA was 3.2%, the bioactivity recovery of a-Chy is the highest.

Isoentropic and Isoenthalpic Temperatures of Protein Unfolding in Hydrophobic Interaction Chromatography

The thermal behaviors of five proteins in hydrophobic interaction chromatography （HIC） were investigated in the temperature range from 0 to 50℃. The thermodynamic parameters （△H°,△S°, △Cp°and △G°） of these proteins in the process of retention and unfolding were determined. The existence of enthalpy and entropy convergence with temperature was confirmed. The differences of the isoentropic and isoenthalpic temperatures for protein unfolding in HIC system from the traditional solution were elucidated.

Resolution enhancement in hydrophobic interaction chromatography via electrostatic interactions

In this work,a new type of hydrophobic stationary phase that provide electrostatic interactions with analytes was developed by bondingβ-phenylethylamine as a functional ligand to silica.This stationary phase can separate proteins with similar hydrophobicity that traditional hydrophobic resins cannot.Hen egg white was separated to examine the selectivity.The results show that the introduced electrostatic interactions are an important factor for the resolution enhancement and the new resin could have important applications in separation and purification of biological macromolecules.

Blue light-induced rare-earth free phosphors for the highly sensitive and selective imaging of latent fingerprints based on enhanced hydrophobic interaction

The demand for in-situ detection of latent fingerprints(LFPs)in ways of high sensitivity,high selectivity,high contrast,low cost and user-friendly is still urgent.To overcome this challenge,a moisture-stable,red-emitting fluoride phosphor K_(3)AlF_(6):Mn^(4+)(KAF:Mn^(4+))with an organic hydrophobic skin was prepared.The phosphor has a uniform and superfine morphology with excellent luminescence properties.More importantly,this non-ultraviolet(UV)or non-near infrared(NIR)induced phosphor was proved to be an ideal fluorescent label for LFP imaging,which is both friendly for touch DNA analysis and compatible to forensic light sources.The well-defined ridge details with little background interference on various surfaces were presented by the oleic acid(OA)modified KAF:Mn^(4+)(KAF:Mn^(4+)-OA)phosphor in few seconds using the powder dusting method.To confirm the high selectivity of KAF:Mn^(4+)-OA for LFP imaging,an efficient quantitative evaluation method is proposed with the aid of ImageJ&Origin software.Due to the superiority of the Mn^(4+)-doped fluoride for the rapid imaging of LFPs in terms of lowcost,high compatibility and good availability,it is expected to be a promising candidate for forensic science as well as fluorescence imaging in other fields instead of rare earth luminescent materials.

Effect of Different Salts on the Solubilities of Benzene and Diphenyl in t-Butyl Alcohol-Water Mixture and Hydrophobic Interaction

The solubilities of benzene and diphenyl in mixed solvents of t-butyl alcohol(TBA) and water with different salts have been determined at T = 298.15, 303.15, 308.15 and 313.15K. The molar fraction of TBA [x(TBA)] in mixed solvent is 0.045, and the molality of the salts (m_s)in mixed solvents are 0.000, 0.250, 0.500, 0.750 and 1.000 mol/kg, respectively. The standard Gibbsenergies of solution of benzene and diphenyl in the mixed solvents have also been calculated basedon the solubility data. The effects of different salts on the hydrophobic interaction (HI) forbenzene-benzene pair in the systems were discussed.

Mechanism of simultaneously refolding and purification of proteins by hydrophobic interaction chromatographic unit and applications

The hydrophobic amino acid residues of a denatured protein molecule tend to react with the particles of the stationary phase of hydrophobic interaction chromatography (STHIC). These hydrophobic interactions prevent the denatured protein molecules from aggregating with each other. The STHIC can provide high enough energy to a denatured protein molecule to make it dehydration and to refold it into its native or various intermediate states. The outcome not only depends on the specific interactions between amino acids, the structure of STHIC, but also depends on the association between the STHIC and mobile phase. The mechanism of protein refolding and the principle of its quality control by HPHIC were also presented. By appropriate selection of the Chromatographie condition, several denatured proteins can be refolded and separated simultaneously in a single Chromatographic run. A specially designed unit, with diameter much larger than its length, was designed and employed for both laboratory and preparative scales. That unit for the simultaneous renaturation and purification of proteins (USRPP) had the following four functions: to completely remove denaturant, to renature proteins, to separate renatured proteins from impurities, and to easily recycle waste denaturant. The efficiencies of refolding and purification of proteins by the USRPP are almost comparable to a usual long Chromatographie column in laboratory. In preparative scale, USRPP can be easily, rapidly, and economically applied requiring a low pressure gradient. As an example, recombinant human interferon-γ is employed to elucidate the application of the preparative USRPP.

Construction of PS/PNIPAM core-shell particles and hollow spheres by using hydrophobic interaction and thermosensitive phase separation

This paper reports an easy and effective way to fabricate polystyrene/poly(N-isopropylacrylamide)(PS/PNIPAM)core-shell particles and PNIPAM hollow spheres.The main point of the method is to take advantage of the hydrophobic interaction between initiator and PS particles.The hydrophobic azodiisobutyronitriles automatically concentrate around the PS particles and initiate polymerization of N-isopropylacrylamide(NIPAM)and the crosslinkermethylene bisacrylamide(MBA),which dissolve in the aqueous phase,at the surface of the PS nanoparticles.Then,PNIPAM adheres to the PS particles to form a coreshell structure as a result of their hydrophobic interaction.This interaction is due to the unique property of PNIPAM,namely,its ability to transition from hydrophilic to hydrophobic when the temperature rises to 32℃.Furthermore,the hollow PNIPAM spheres were obtained by etching the PS core with chloroform.

Effect of hydrophilic or hydrophobic interactions on the self-assembly behavior and micro-morphology of a collagen mimetic peptide

Peptide self-assembles with bionic properties have been widely utilized for bioactive drugs and biomedical materials.Collagen mimetic peptide(CMP)gains more attention due to its unique advantages in biosecurity and function.Unfortunately,the self-assembly mechanism of CMP,particularly the effect of intermolecular forces on its self-assembly behavior and morphology,is still unrecognized.Herein,the hydrophilic glycidol(GCD)and hydrophobic Y-glycidyl ether oxypropyl trimethoxysilane(GLH)were grafted onto the side chains of CMP through the ring-opening reaction(GCD/CMP,GLH/CMP).Subsequently,the effects of hydrophilic and hydrophobic interactions on the self-assembly behavior and morphology of CMP were further studied.The results substantiated that the GCD/CMP and GLH/CMP self-assembly followed“nucleation-growth”mechanism,and the supererogatory hydrophilic and hydrophobic groups prolonged the nucleation and growth time of CMP self-assembly.Noted that the hydrophilic interaction had stronger driving effects than hydrophobic interaction on the self-assembly of CMP.The GCD/CMP and GLH/CMP self-assembles exhibited fibrous 3D network and microsphere morphology,respectively.Furthermore,the GLH/CMP self-assembles had better resistance to degradation.Consequently,the microtopography and degradation properties of CMP self-assembles could be controlled by the hydrophilic and hydrophobic interactions between CMP,which would further provide a way for subsequent purposeful design of biomedical materials.

Solubilities of 1,2,3-trimethylbenzene and 1,2,4-trimethylbenzene and 1,3,5-trimethylbenzene in t-butyl alcohol + water mixtures and hydrophobic interaction

The solubilities of 1,2,3-trimethylbenzene, 1,2,4-trimethylbenzene and 1,3,5-trimethylbenzene in mixed solvents of t-butyl alcohol (TBA) and water at 283.15, 288.15, 293.15 and 298.15 K have been determined by spectmphotometry. The mole fractions of TBA [x(TBA)] in the mixed solvent are0.000, 0.010, 0.020, 0.030, 0.040, 0.045, 0.050, 0.060, 0.080 and 0.1000. The Gibbs energies of hydrophobic interaction (HI) for the aggregating process of three methane molecules with one benene molecule in the mixed solvent are studied, and the effect of solvent structure and solute aggregating state on the strength of HI is discussed.

Hydrophobic interaction membrane chromatography for bioseparation and responsive polymer ligands involved

Hydrophobic interaction chromatography （HIC） is a rapid growing bioseparation technique, which separates biomolecules, such as therapeutic proteins and antibodys, based on the reversible hydrophobic interaction between immobilized hydrophobic ligands on chromatographic resin spheres and non-polar regions of solute molecule. In this review, the fundamental concepts of HIC and the factors that may affect purification efficiency of HIC is summarized, followed by the comparison of HIC with affinity chromatography and ion-exchange chromatography. Hydrophobic interaction membrane chromatography （HIMC） combines the advantages of HIC and membrane process and has showed great potential in bioseparation. For better understanding of HIMC, this review presents an overview of two main concerns about HIMC, i.e. membrane materials and hydrophobic ligands. Specifically, cellulose fiber-based membrane substrate and environment-responsive ligands are emphasized.

Effect of ligand structure of stationary phase of high performance hydrophobic interaction chromatography on renaturation efficiency of GuHCl-denaturedα-chymotrypsin

The renaturation of the denaturedα-chymotrypsin(α-Chy)with 1.7 mol·L^(-1)guanidine hydrochloride(GuHCI)by three kinds of stationary phase of high performance hydrophobic interaction chromatography(STHIC)with a comparable hydrophobicity but different ligand structures was investigated.The obtained result indicates that the ligand structures of the three STHIC contribute to the renaturation efficiency ofα-Chy in the order of the end ligands PEG-600<phenyl group<tetrahydrofurfuryl alcohol(THFA).

Enhancement of intramolecular excimer formation and photodimerization of anthrylmethyl α,ω-alkanedioates via hydrophobic interactions

The fluorescence spectra and photodimerization of anthrylmethyl a,w-alkanedioates (A-Mn-A) both in organic and in aqueous organic mixed solvents have been studied.In aqueous organic mixed solvents strong intramolecular excimer emission is detected and the quantum yield for the intramolecular photodimerization is significantly greater than those in organic solvents.These observations suggest that hydrophobic interactions force A-Mn-A molecule to self-coil.The ratio of the head-to-head to head-to-tail products in the intramolecular photodimers of A-Mn-A depends on the length of the linking chain.This work presents a successful example of application of hydrophobic interactions to enhancement of large-ring formation.

Retention model of protein for mixed-mode interaction mechanism in ion exchange and hydrophobic interaction chromatography

A unified retention equation of proteins was proved to be valid for a mixed-mode interaction mechanism in ion exchange chromatography (IEC) and hydrophobia interaction chro-matography (HIC). The reason to form a 'U' shape retention curve of proteins hi both HIC and IEC was explained and the concentration range of the strongest elution ability for the mobile phase was determined with this equation. The parameters in this equation could be used to characterize the difference for either HIC or IEC adsorbents and the changes in the molecular conformation of proteins. With the parameters in this equation, the contributions of salt and water in the mobile phase to the protein retention in HIC and IEC were discussed, respectively. In addition, the comparison between the unified equation and Melander' s three-parameter equation for mixed-mode interaction chromatography was also investigated and better results were obtained in former equation.

Synthesis of Novel Hydrophobic Media and Purification of Recombinant HBsAg

A novel hydrophobic medium with propyl as functional group and Sepharose 6B as matrix was designed and synthesized. The comparison of the hydrophobic medium synthesized with the commercial products was made by hydrophobic interaction chromatography(HIC) in isolating recombinant hepatitis B surface antigen(r HBsAg). r HBsAg was further purified to the final products by following a downstream procedure . The results indicate that the synthesized hydrophobic medium possesses a stable structure and desired physical and chemical properties. They were used to purify r HBsAg with a high yield and purity. Both the immunity and stability of hepatitis vaccine made by the r HBsAg products have reached the same level as other similar kinds of products.

Neural Network Learning of the Interaction Between Peptide Segments of Proteins

neural network model based on backbone propagation was applied to Learn-ing and predicting the interaction between antiparallelly interactive peptide seg-ments in proteins.Hydrophobic properties pf residues were found dominant in in-terpeptides.Weights of each kind of residues, obtained by this work,suggestedsome different scales for the hydrophobicity of the residue.These will be helpful in understanding polypeptide structure and protein folding.

Physico-chemical parameters for the assembly of moxifloxacin hydrochloride and cetyltrimethylammonium chloride mixture in aqueous and alcoholic media

The aggregation behavior of the mixture of cetyltrimethylammonium chloride(CTAC), a cationic surfactant, and moxifloxacin hydrochloride(MFH), a fourth-generation fluoroquinolone antibiotic drug, has been studied using the conductivity technique in aqueous and alcoholic(EtOH, 1-PrOH, and 2-BuOH)media. The study was performed at several temperatures between 298.15 and 323.15 K at 5 K intervals.The assembly has been characterized by evaluating the micellar parameters, such as the critical micelle concentration(CMC) and the counter ion binding(β), of the CTAC + MFH mixture. The values of the CMC for the assembly of the CTAC + MFH mixture were reliant on the composition of alcohols in the mixed solvents and the temperature. The CMC values of the CTAC + MFH mixture increased with increasing temperature;that is, assembly was delayed by increased temperature. The micellization of the CTAC + MFH mixed system was delayed in alcoholic media. The observed-ΔG0mvalues for the association of the CTAC + MFH mixed system demonstrated a spontaneous aggregation process under all study conditions.Based on the-ΔH^(0)_(m) and +ΔS^(0)_(m) values, the association of the CTAC + MFH mixture is exothermic and the interaction forces acting between the CTAC and MFH species are hydrophobic, ion–dipole, and electrostatic interactions. The transfer properties and enthalpy–entropy compensation were also assessed and described comprehensively.

Effect of Potassium Oleate on Rheological Behavior of Cationic Guar in Aqueous Solution with Varying Temperatures

The rheology of the cationic guar （CG） solution was measured and the effects of potassium oleate （KOA） upon the rheological properties of CG solution were studied. The steady shear viscosity measurement has shown that the viscosity of CG solution increased dramatically in the presence of KOA. The viscosity enhancement of KOA upon CG solution can be approximate three orders in magnitude. The gel-like formation of CG solution is observed at the high concentration of KOA. The excess addition of KOA results in the phase separation of CG solution. The oscillatory rheological measurement has shown that the crossover modulus Gc （corresponding to either storage modulus G＇ or loss modulus G＇＇ at the frequency wc where G＇ equals G＇＇） for CG solution, decreases with the increasing the concentration of KOA in solution. On the other hand, the apparent relaxation time 7-app （=1/wc） increases with increasing the concentration of KOA in solution. Our experimental results suggest that for surfaetant such as KOA which has a stronger tendency to form micelles in solution, the cooperative hydrophobic interaction of polymer bound to surfactants is less necessary to the formation of aggregates in solution, especially at the high concentration of surfactants. In fact, with the increase of the concentration of KOA, the number of the aggregates which associate polymer together decreases whereas the intensity of these aggregates increases. The effect of temperature upon the aggregation is also significant. With the increase of temperature, the number of the aggregates increases whereas the intensity of these aggregates decreases, probably because the ionization of KOA increases at high temperature.

Adsorption Mechanisms of Mesoporous Adsorbents in Solutions

Sieve effect, complexation, ionic exchange, electrostatic interaction, hydrogen bonding, hydrophobic interaction, and molecular recognition based on molecular imprinting are comprehensively discussed.

Effect of Short Chain Alcohols upon Viscosity of TTAB Solution

The effect of ethanol （C2H5OH）, propanol （C3H7OH）, and butanol （C4H9OH） upon the viscosity of tetradecyltrimethylammonium bromide （TTAB） solution in the presence or absence of KBr at 30℃was investigated, where the surfactant concentration CS is kept constant. In the absence of KBr, the relative viscosity ηr of TTAB solution increases linearly with the alcohol concentration CA, indicating that the alcohols do not promote micelle formation of TTAB. In the presence of KBr, ηr linearly decreases with CA for C2H5OH, but it exhibits a maximum with increasing CA for C3H7OH or C4H9OH. The facts reveal that C2H5OH or C4H9OH promotes the micelle formation of TTAB. A possible explanation is that the hydrophobicity of the mieellar interior is enhanced by KBr, so that C2H5OH or C4H9OH can dissolve in micelle and promotes micelle formation. In the presence of KCl, which is less efficient in promoting the micelle formation of cationic surfactant, both C3H7OH and C4H9OH have only a slight effect on the micelle formation. In contrast, due to the hydrophilicity, C2H5OH cannot dissolve in micelles in the presence of KBr or KCl.