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Solution Conditions (solution + condition)
Selected AbstractsBiomimetic Silica Formation: Effect of Block Copolypeptide Chemistry and Solution Conditions on Silica Nanostructure,ADVANCED MATERIALS, Issue 19 2007J.-S. Jan Biomimetic syntheses of silicas using block copolypeptides containing a hydrophilic lysine block are reported. Porous silica platelets or spheres can be made using block copolypeptides as templates. The results show that biomimetic synthesis under benign conditions may be a potential route for assembling nanostructured materials. [source] Role of Iron(III) and Aluminum Hydroxides in Concentration/reduction of Au(III) ComplexesRESOURCE GEOLOGY, Issue 3 2002Akiko UCHIDA Abstract: The adsorption of gold on iron(III) and aluminum hydroxides from solutions containing Au(III) complexes has been studied as a function of pH and chloride concentration at 30C. Iron(III) hydroxide was more effective in adsorbing gold from solution than aluminum hydroxide. However, both hydroxides controlled the behavior of Au(III) complex with very similar manner. The most effective gold adsorption occurred in aqueous solution with near neutral pH and low Cl concentration. In this solution condition, Au(III) complexes were mainly dissolved as AuCl2(OH)2 - and AuCl(OH)3 - , and the surface charge for both hydroxides was positive. In addition, the adsorbed Au(III) complexes were spontaneously reduced to elemental gold in spite of the absence of a specific reducing agent. The results of this study suggest that adsorption and spontaneous reduction of gold complexes on the surface of hydrous metal oxides with positive charge play an important role in gold precipitation in subsurface environment. [source] Particle size of powders under hydrothermal conditionsCRYSTAL RESEARCH AND TECHNOLOGY, Issue 10 2003Wen-Jun Li Abstract Various non-oxide (CuI, AgI, AgCl, PbS, CuS and ZnS) and oxide (ZnO, TiO2, SnO2, CeO2 and ZrO2) powders were prepared under hydrothermal conditions to investigate the effects of temperature, pH and precursors on the particle size of powders. It was found that the particle sizes of PbS, CuS and ZnS powders were much smaller than that of CuI, AgI and AgCl powders prepared under the same conditions. The particle sizes of TiO2, SnO2, CeO2 and ZrO2 powders are much smaller than that of ZnO powders prepared under the same conditions. It is concluded that the solution conditions have a certain effect on the particle size of powders under the hydrothermal conditions. The particle size of powders increased with the rising of temperature. Additional factors affecting the particle size were uncovered through studying the nucleation mechanism. The particle size was mainly related to the Madelung constant and the electric charge number of ions. Powders with smaller particle size resulted from systems that possessed the larger Madelung constant and ionic charge number. (© 2003 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Metalloporphyrin solubility: A trigger for catalyzing reductive dechlorination of tetrachloroethyleneENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 2 2004Ishai Dror Abstract Metalloporphyrins are well known for their electron-transfer roles in many natural redox systems. In addition, several metalloporphyrins and related tetrapyrrole macrocycles complexed with various core metals have been shown to catalyze the reductive dechlorination of certain organic compounds, thus demonstrating the potential for using naturally occurring metalloporphyrins to attenuate toxic and persistent chlorinated organic pollutants in the environment. However, despite the great interest in reductive dechlorination reactions and the wide variety of natural and synthetic porphyrins currently available, only soluble porphyrins, which comprise a small fraction of this particular family of organic macrocycles, have been used as electron-transfer shuttles in these reactions. Results from the present study clearly demonstrate that metalloporphyrin solubility is a key factor in their ability to catalyze the reductive dechlorination of tetrachloroethylene and its daughter compounds. Additionally, we show that certain insoluble and nonreactive metalloporphyrins can be activated as catalysts merely by changing solution conditions to bring about their dissolution. Furthermore, once a metalloporphyrin is fully dissolved and activated, tetrachloroethylene transformation proceeds rapidly, giving nonchlorinated and less toxic alkenes as the major reaction products. Results from the present study suggest that if the right environmental conditions exist or can be created, specific metalloporphyrins may provide a solution for cleaning up sites that are contaminated with chlorinated organic pollutants. [source] Application of ultrasonic shear rheometer to characterize rheological properties of high protein concentration solutions at microliter volumeJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 6 2005Atul Saluja Abstract The purpose of this work was to conduct preliminary rheological analysis on high protein concentration solutions by using the technique of ultrasonic shear rheometry at megahertz frequencies. The work was aimed at establishing the viability of the technique for analyzing protein solution rheology as well as obtaining an initial understanding of the effect of solution conditions on solution rheology of a model protein. Bovine serum albumin (BSA) was used for this study, and rheological analysis was conducted at 20 ,L sample volume between pH 2.0 and 9.0 at different ionic strengths at 25°C using 5 and 10 MHz quartz crystals. Significant differences in storage modulus among solutions at pH 5.0, 7.0, and 9.0 could only be detected at 10 MHz, and the errors associated with measurements were smaller as compared to those at 5 MHz for all the solutions studied. Solutions at pH 2.0 and 3.0 showed a time-dependent change in solution rheology. For solutions at pH 5.0, 7.0, and 9.0, which did not show time dependence in solution rheology, loss modulus data at lower concentrations correlated well with the dilute solution data in the literature. At higher concentrations, pH 5.0 solutions exhibited a higher loss modulus than pH 7.0 and pH 9.0 solutions. Storage modulus decreased with increasing ionic strength, unlike loss modulus, which did not show any change, except at pI of protein when no effect was observed. The results show the potential of high frequency rheometry for analyzing subtle differences in rheology of pharmaceutically relevant protein solutions at microliter volume. © 2005 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 94:1161,1168, 2005 [source] Nanocrystalline Tin Oxide Thin Films via Liquid Flow DepositionJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 12 2003Sitthisuntorn Supothina Nanocrystalline films of SnO2 were deposited by liquid flow deposition (LFD), i.e., by flowing aqueous solutions of SnCl4·5H2O and HCl over single-crystalline silicon substrates at 80°C. The substrates were either oxidized and fully hydrolyzed (bare silicon) or oxidized, hydrolyzed, and then coated with siloxy-anchored organic self-assembled monolayers (SAMs). Continuous, adherent films formed on sulfonate- and thioacetate-functionalized SAMs; adherent but sometimes discontinuous films formed on bare silicon and methyl-functionalized SAMs. The films contained equiaxed cassiterite crystals, ,4,10 nm in size. The film thickness increased linearly with deposition time. The maximum growth rate observed was 85 nm·h,1 on sulfonate SAM, and the maximum film thickness obtained was 1 ,m. A new dimensionless parameter, the normalized residence time, ,, was introduced for the purpose of interpreting the influence of solution conditions (i.e., degree of supersaturation, as controlled via pH, and tin concentration) and flow characteristics (flow rate and the configuration of the deposition chamber) on the growth rate in LFD processes. The results were consistent with a particle attachment mechanism for film growth and inconsistent with heterogeneous nucleation on the substrate. [source] Assembly of Polyaniline NanostructuresMACROMOLECULAR RAPID COMMUNICATIONS, Issue 1 2007Jixiao Wang Abstract Polyaniline nanostructures (nanosheets, nanofibers, and nanoparticles) can be assembled at the organic/aqueous interface or in solution by controlling the diffusion rate and the polymerization induction time of aniline. The quality of polyaniline nanostructures is determined by the polymerization solution conditions. Polyaniline nanosheets formation mechanism was proposed. Under certain polymerization conditions, polyaniline nanofibers or/and nanoparticles were obtained. [source] Preparation and characterization of ultrafine electrospun polyacrylonitrile fibers and their subsequent pyrolysis to carbon fibersPOLYMER INTERNATIONAL, Issue 8 2006Juthawan Sutasinpromprae Abstract The present contribution reports the fabrication and characterization of ultrafine polyacrylonitrile (PAN) fibers by electrospinning and further development of the as-spun PAN fibers into ultrafine carbon fibers. The effects of solution conditions (i.e., solution concentration, viscosity, conductivity, and surface tension) and process parameters (i.e., applied electrostatic field strength, emitting electrode polarity, nozzle diameter, and take-up speed of a rotating-drum collector) on morphological appearance and average diameter of the as-spun PAN fibers were investigated by optical scanning (OS) and scanning electron microscopy (SEM). The concentration, and hence the viscosity, of the spinning solutions significantly affected the morphology and diameters of the as-spun PAN fibers. The applied electrostatic field strength and nozzle diameter slightly affected the diameters of the as-spun fibers, while the emitting electrode polarity did not show any influence over the morphology and size of the as-spun fibers. Utilization of the rotating-drum collector enhanced the alignment of the as-spun fibers. Within the investigated concentration range, the average diameter of the fibers ranged between 80 and 725 nm. Finally, heat treatment of the as-spun fibers with their average diameter of about 450 nm was carried out at 230 and 1000 °C, respectively. Various characterization techniques revealed successful conversion into carbon fibers with an average diameter of about 250 nm. Copyright © 2006 Society of Chemical Industry [source] Effects of additives on surfactant phase behavior relevant to bacteriorhodopsin crystallizationPROTEIN SCIENCE, Issue 12 2006Bryan W. Berger Abstract The interactions leading to crystallization of the integral membrane protein bacteriorhodopsin solubilized in n-octyl-,-D-glucoside were investigated. Osmotic second virial coefficients (B22) were measured by self-interaction chromatography using a wide range of additives and precipitants, including polyethylene glycol (PEG) and heptane-1,2,3-triol (HT). In all cases, attractive protein,detergent complex (PDC) interactions were observed near the surfactant cloud point temperature, and there is a correlation between the surfactant cloud point temperatures and PDC B22 values. Light scattering, isothermal titration calorimetry, and tensiometry reveal that although the underlying reasons for the patterns of interaction may be different for various combinations of precipitants and additives, surfactant phase behavior plays an important role in promoting crystallization. In most cases, solution conditions that led to crystallization fell within a similar range of slightly negative B22 values, suggesting that weakly attractive interactions are important as they are for soluble proteins. However, the sensitivity of the cloud point temperatures and resultant coexistence curves varied significantly as a function of precipitant type, which suggests that different types of forces are involved in driving phase separation depending on the precipitant used. [source] Expression, purification, and characterization of Thermotoga maritima membrane proteins for structure determinationPROTEIN SCIENCE, Issue 5 2006Linda Columbus Abstract Structural studies of integral membrane proteins typically rely upon detergent micelles as faithful mimics of the native lipid bilayer. Therefore, membrane protein structure determination would be greatly facilitated by biophysical techniques that are capable of evaluating and assessing the fold and oligomeric state of these proteins solubilized in detergent micelles. In this study, an approach to the characterization of detergent-solubilized integral membrane proteins is presented. Eight Thermotoga maritima membrane proteins were screened for solubility in 11 detergents, and the resulting soluble protein,detergent complexes were characterized with small angle X-ray scattering (SAXS), nuclear magnetic resonance (NMR) spectroscopy, circular dichroism (CD) spectroscopy, and chemical cross-linking to evaluate the homogeneity, oligomeric state, radius of gyration, and overall fold. A new application of SAXS is presented, which does not require density matching, and NMR methods, typically used to evaluate soluble proteins, are successfully applied to detergent-solubilized membrane proteins. Although detergents with longer alkyl chains solubilized the most proteins, further characterization indicates that some of these protein,detergent complexes are not well suited for NMR structure determination due to conformational exchange and protein oligomerization. These results emphasize the need to screen several different detergents and to characterize the protein,detergent complex in order to pursue structural studies. Finally, the physical characterization of the protein,detergent complexes indicates optimal solution conditions for further structural studies for three of the eight overexpressed membrane proteins. [source] Temperature-induced reversible conformational change in the first 100 residues of ,-synucleinPROTEIN SCIENCE, Issue 3 2006Brian C. McNulty Abstract Natively disordered proteins are a growing class of anomalies to the structure,function paradigm. The natively disordered protein ,-synuclein is the primary component of Lewy bodies, the cellular hallmark of Parkinson's disease. We noticed a dramatic difference in dilute solution 1H- 15N Heteronuclear Single Quantum Coherence (HSQC) spectra of wild-type ,-synuclein and two disease-related mutants (A30P and A53T), with spectra collected at 35°C showing fewer cross-peaks than spectra acquired at 10°C. Here, we show the change to be the result of a reversible conformational exchange linked to an increase in hydrodynamic radius and secondary structure as the temperature is raised. Combined with analytical ultracentrifugation data showing ,-synuclein to be monomeric at both temperatures, we conclude that the poor quality of the 1H- 15N HSQC spectra obtained at 35°C is due to conformational fluctuations that occur on the proton chemical shift time scale. Using a truncated variant of ,-synuclein, we show the conformational exchange occurs in the first 100 amino acids of the protein. Our data illustrate a key difference between globular and natively disordered proteins. The properties of globular proteins change little with solution conditions until they denature cooperatively, but the properties of natively disordered proteins can vary dramatically with solution conditions. [source] Chaperonin-assisted folding of glutamine synthetase under nonpermissive conditions: Off-pathway aggregation propensity does not determine the co-chaperonin requirementPROTEIN SCIENCE, Issue 12 2000Paul A. Voziyan Abstract One of the proposed roles of the GroEL-GroES cavity is to provide an "infinite dilution" folding chamber where protein substrate can fold avoiding deleterious off-pathway aggregation. Support for this hypothesis has been strengthened by a number of studies that demonstrated a mandatory GroES requirement under nonpermissive solution conditions, i.e., the conditions where proteins cannot spontaneously fold. We have found that the refolding of glutamine synthetase (GS) does not follow this pattern. In the presence of natural osmolytes trimethylamine N-oxide (TMAO) or potassium glutamate, refolding GS monomers readily aggregate into very large inactive complexes and fail to reactivate even at low protein concentration. Surprisingly, under these "nonpermissive" folding conditions, GS can reactivate with GroEL and ATP alone and does not require the encapsulation by GroES. In contrast, the chaperonin dependent reactivation of GS under another nonpermissive condition of low Mg2+ (<2 mM MgCl2) shows an absolute requirement of GroES. High-performance liquid chromatography gel filtration analysis and irreversible misfolding kinetics show that a major species of the GS folding intermediates, generated under these "low Mg2+" conditions exist as long-lived metastable monomers that can be reactivated after a significantly delayed addition of the GroEL. Our results indicate that the GroES requirement for refolding of GS is not simply dictated by the aggregation propensity of this protein substrate. Our data also suggest that the GroEL-GroES encapsulated environment is not required under all nonpermissive folding conditions. [source] Progress in rational methods of cryoprotection in macromolecular crystallographyACTA CRYSTALLOGRAPHICA SECTION D, Issue 4 2010Thomas Alcorn Cryogenic cooling of macromolecular crystals is commonly used for X-ray data collection both to reduce crystal damage from radiation and to gather functional information by cryogenically trapping intermediates. However, the cooling process can damage the crystals. Limiting cooling-induced crystal damage often requires cryoprotection strategies, which can involve substantial screening of solution conditions and cooling protocols. Here, recent developments directed towards rational methods for cryoprotection are described. Crystal damage is described in the context of the temperature response of the crystal as a thermodynamic system. As such, the internal and external parts of the crystal typically have different cryoprotection requirements. A key physical parameter, the thermal contraction, of 26 different cryoprotective solutions was measured between 294 and 72,K. The range of contractions was 2,13%, with the more polar cryosolutions contracting less. The potential uses of these results in the development of cryocooling conditions, as well as recent developments in determining minimum cryosolution soaking times, are discussed. [source] The Fouling Behaviour of Sodium Aluminosilicate Polytypes in High Ionic Strength Caustic Media Heat ExchangersASIA-PACIFIC JOURNAL OF CHEMICAL ENGINEERING, Issue 1-2 2006J. Addai-Mensah Sodium aluminosilicate (SAS) scale formation, simulating high ionic strength caustic media process heat exchanger fouling, has been investigated for a temperature range of 30,240°C. Findings from several studies performed with a variety of liquor, substrate-type and agitation conditions show that SAS polytypes; i.e. amorphous, zeolite A, sodalite and cancrinite phases, differing in thermodynamic stability and kinetic behaviour may deposit. The polytypes formed and subsequent transformations to more thermodynamically stable phases are strongly dependent on liquor composition, temperature and time. The scale deposition process is substrate- mediated heterogeneous nucleation and particle growth (precipitation foulin&, which may be accompanied by bulk liquor-nucleated solids adsorption (particulate fouling) at suficiently high supersaturations. Liquor seeding with stable SAS solid phases is found to be efective in mitigating fouling. The precipitation fouling behaviour is principally determined by the clystallo-chemical nature of the SAS polytype and the solution conditions. The substrate (mild steel, 316 stainless steel, nickel and tef2on) surface physico-chemical structure has a signlficant impact on the scale particle morphology and layer structure, particularly at high temperatures. [source] Conformational transition and liquid crystalline state of regenerated silk fibroin in waterBIOPOLYMERS, Issue 6 2008Xin-Gui Li Abstract The conformational transition of molecular chains of regenerated silk fibroin (SF) aqueous solution is systematically investigated by circular dichroism, Raman, IR, and UV,vis spectroscopies. It is found that an initial random coil conformation of the SF can be readily changed into an ordered ,-sheet structure by optimizing the solution conditions, such as the SF concentration, pH, temperature, or metal-ion content. Circular dichroic spectra quantitatively confirm a steadily decreased content of the random coil conformation but a significantly increased ,-sheet content after an ultrasonic or extruding treatment. Furthermore, the extrusion is more powerful to achieve high ,-sheet content than the ultrasonic. It is interesting that the polarized optical micrographs of the SF aqueous solution extruded by injection illustrate the formation and existence of liquid crystalline state. A study of extrusion in vitro could be used as a model system to understand the natural silk spinning process in silkworm. © 2007 Wiley Periodicals, Inc. Biopolymers 89: 497,505, 2008. This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com [source] Lysozyme-lysozyme self-interactions as assessed by the osmotic second virial coefficient: Impact for physical protein stabilizationBIOTECHNOLOGY JOURNAL, Issue 9 2009Virginie Le Brun Abstract The purpose of the presented study is to understand the physicochemical properties of proteins in aqueous solutions in order to identify solution conditions with reduced attractive protein-protein interactions, to avoid the formation of protein aggregates and to increase protein solubility. This is assessed by measuring the osmotic second virial coefficient (B22), a parameter of solution non-ideality, which is obtained using self-interaction chromatography. The model protein is lysozyme. The influence of various solution conditions on B22 was investigated: protonation degree, ionic strength, pharmaceutical relevant excipients and combinations thereof. Under acidic solution conditions B22 is positive, favoring protein repulsion. A similar trend is observed for the variation of the NaCl concentration, showing that with increasing the ionic strength protein attraction is more likely. B22 decreases and becomes negative. Thus, solution conditions are obtained favoring attractive protein-protein interactions. The B22 parameter also reflects, in general, the influence of the salts of the Hofmeister series with regard to their salting-in/salting-out effect. It is also shown that B22 correlates with protein solubility as well as physical protein stability. [source] A rapid, sensitive and economical assessment of monoclonal antibody conformational stability by intrinsic tryptophan fluorescence spectroscopyBIOTECHNOLOGY JOURNAL, Issue 9-10 2008Patrick Garidel Dr. Abstract Steady-state intrinsic tryptophan fluorescence spectroscopy is used as a rapid, robust and economic way for screening the thermal protein conformational stability in various formulations used during the early biotechnology development phase. The most important parameters affecting protein stability in a liquid formulation, e. g. during the initial purification steps or preformulation development, are the pH of the solution, ionic strength, presence of excipients and combinations thereof. A well-defined protocol is presented for the investigation of the thermal conformational stability of proteins. This allows the determination of the denaturation temperature as a function of solution conditions. Using intrinsic tryptophan fluorescence spectroscopy for monitoring the denaturation and folding of proteins, it is crucial to understand the influence of different formulation parameters on the intrinsic fluorescence probes of proteins. Therefore, we have re-evaluated and re-assessed the influence of temperature, pH, ionic strength, buffer composition on the emission spectra of tryptophan, phenylalanine and tyrosine to correctly analyse and evaluate the data obtained from thermal-induced protein denaturation as a function of the solution parameters mentioned above. The results of this study are a prerequisite for using this method as a screening assay for analysing the conformational stability of proteins in solution. The data obtained from intrinsic protein fluorescence spectroscopy are compared to data derived from calorimetry. The advantage, challenges and applicability using intrinsic tryptophan fluorescence spectroscopy as a routine development method in pharmaceutical biotechnology are discussed. [source] Effect of antibody solution conditions on filter performance for virus removal filter PlanovaÔ 20NBIOTECHNOLOGY PROGRESS, Issue 4 2010Tomoko Hongo-Hirasaki Abstract We investigated the effect of antibody solution conditions (ionic strength, pH, IgG concentration, buffer composition, and aggregate level (dimer content)) on filter performance for a virus removal filtration process using the PlanovaÔ 20N, a virus removal filter. Ionic strength and pH affected the filter flux. A consistent high flux was maintained at an ionic strength greater than 10 mM and at pH 4,8 under a typical buffer composition (sodium chloride, citrate, acetate, and phosphate). Optimum IgG concentration was 10,20 mg/mL allowing for high throughput (kg/m2 of IgG). Dimer content negligibly affected the flux level. Under high throughput conditions, virus spiking did not affect flux whereas a parvovirus logarithmic reduction value greater than 5 was maintained. From the results of zeta potential analyses for IgG and the membrane, we considered that electrostatic interactions between antibodies and the membrane affect filter performance (flux level and throughput). These results indicate that the PlanovaÔ 20N filter is applicable for a wide range of solution conditions typically used in antibody processing. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source] Probing protein colloidal behavior in membrane-based separation processes using spectrofluorometric Rayleigh scattering dataBIOTECHNOLOGY PROGRESS, Issue 3 2010Rand Elshereef Abstract One of the primary problems in membrane-based protein separation is membrane fouling. In this study we explored the feasibility of employing Rayleigh light scattering data from fluorescence studies combined with chemometric techniques to determine whether a correlation could be established with membrane fouling phenomena. Membrane flux was measured in a dead-end UF filtration system and the effect of protein solution properties on the flux decline was systematically investigated. A variety of proteins were used as a test case in this study. In parallel, the colloidal behavior of the protein solutions was assessed by employing multiwavelength Rayleigh scattering measurements. To assess the usefulness of Rayleigh scattering measurements for probing the colloidal behavior of proteins, a protein solution of ,-lactoglobulin was used as a base-case scenario. The colloidal behavior of different ,-lactoglobulin solutions was inferred based on published data for this protein, under identical solution conditions, where techniques other than Rayleigh scattering had been used. Using this approach, good agreement was observed between scattering data and the colloidal behavior of this protein. To test the hypothesis that a high degree of aggregation will lead to increased membrane fouling, filtration data was used to find whether the Rayleigh scattering intensity correlated with permeate flux changes. It was found that for protein solutions which were stable and did not aggregate, fouling was reduced and these solutions exhibited reduced Rayleigh scattering. When the aggregation behavior of the solution was favored, significant flux declines occurred and were highly correlated with increased Rayleigh scattering. © 2010 American Institute of Chemical Engineers Biotechnol. Prog., 2010 [source] Factors influencing antibody stability at solid,liquid interfaces in a high shear environmentBIOTECHNOLOGY PROGRESS, Issue 5 2009James G. Biddlecombe Abstract A rotating disk shear device was used to study the effect of interfacial shear on the structural integrity of human monoclonal antibodies of IgG4 isotype. Factors associated with the solution conditions (pH, ionic strength, surfactant concentration, temperature) and the interface (surface roughness) were studied for their effect on the rate of IgG4 monomer loss under high shear conditions. The structural integrity of the IgG4 was probed after exposure to interfacial shear effects by SDS-PAGE, IEF, dynamic light scattering, and peptide mapping by LC-MS. This analysis revealed that the main denaturation pathway of IgG4 exposed to these effects was the formation of large insoluble aggregates. Soluble aggregation, breakdown in primary structure, and chemical modifications were not detected. The dominant factors found to affect the rate of IgG4 monomer loss under interfacial shear conditions were found to be pH and the nanometer-scale surface roughness associated with the solid-liquid interface. Interestingly, temperature was not found to be a significant factor in the range tested (15,45°C). The addition of surfactant was found to have a significant stabilizing effect at concentrations up to 0.02% (w/v). Implications of these findings for the bioprocessing of this class of therapeutic protein are briefly discussed. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source] Screening of Protein-Ligand Interactions by Affinity ChromatographyBIOTECHNOLOGY PROGRESS, Issue 2 2003Carlos D. García This paper examines affinity chromatography (AC) as an alternative tool for the determination of protein-ligand interactions for the particular case in which the ligand is the same protein. The methodology is less labor-intensive and more sample-efficient than traditional methods used to measure the second virial coefficient ( B22), a parameter commonly used to evaluate protein-protein interactions. The chromatographic capacity factor ( k,) was studied for lysozyme and equine serum albumin for a wide range of experimental solution conditions such as crystallizing agent concentration, protein concentration and pH. Parallel experiments using AC to determine k, and static light scattering (SLS) to determine B22 showed that the two parameters were highly correlated. Two different column volumes (,1 and ,0.1 mL) were tested and gave essentially the same values for k,, showing the feasibility of miniaturization. [source] | |||||||||||||||||||