Amount Of Adsorbed Protein Research Articles

Impacts of stereoregularity and stereocomplex formation on physicochemical, protein adsorption and cell adhesion behaviors of star-shaped 8-arms poly(ethylene glycol)–poly(lactide) block copolymer films

Biodegradable stereocomplex film exhibiting soft and stretchy character was prepared by simply blending between enantiomeric 8-arms poly(ethylene glycol)- block-poly( l-lactide) (8-arms PEG- b-PLLA) and 8-arms PEG- b-PDLA copolymers with star-shaped structure. The stereocomplex film exhibited higher T g and PLA crystallinity than those of original copolymer films. Effects of stereoregularity and stereocomplexation on protein adsorption and L929 cells attachment/proliferation behaviors onto the films were analyzed from the viewpoint to design a new class of implantable soft biomaterial. The stereocomplex film was found to exhibit large amount of protein adsorption than original films. Furthermore, cell attachment efficiency and proliferation rate on the film were significantly enhanced by stereocomplexation. This stereocomplex material is expected to be applicable as degradable temporary scaffold for soft tissue regeneration. Consequently, it was indicated that the stereocomplex formation could be proposed to be a novel method to control the protein- and cell-adhesive properties of biodegradable matrix composed of PEG–PLA copolymer.

신규 합성 당지질 함유 리포솜의 In Vitro 안정성

리포솜은 수십~수백 나노미터 크기의 소포체(small vesicle)로서 약물전달에 유용한 구조체이나 체내 혈류 순환계(blood circulatory system)에서 구조적 불안정성에 의해 체내 순환시간이 짧고 세망내피계(reticuloendothelial system)에 의해 소실되어 이를 개선하기 위한 다양한 연구가 시도되고 있다. 본 연구에서는 이당류(disaccharide)로서 락토오스와 슈크로스가 1,2-디스테아로일-sn-글리세로-3-포스포에탄올아민(1,2-Distearoylsn-glycero-3-phosphoethanolamine, DSPE)과 공유결합된 새로운 당-DSPE 유도체를 합성하고 이를 리포솜의 구성성분으로 하는 리포솜을 제조함으로써 리포솜의 표면이 이당으로 수식된 리포솜을 제조하였다. 제조된 리포솜의 입자크기는 대략 100 nm였으며, 표면전하 값은 당이 수식되지 않은 대조군 리포솜이 -10 mV를 나타내었으나 당-DSPE 유도체를 함유한 리포솜의 경우 리포솜 표면에 존재하는 당의 수산화기에 의하여 표면전하의 값은 -25 mV를 나타내었다. 리포솜 내부에 모델약물인 독소루비신(doxorubicin)의 로딩효율은 약 90%를 나타내었다. 당-DSPE 유도체 함유 리포솜의 in vitro 안정성은 혈청 내에서 단백흡착량의 변화 및 리포솜의 입자크기를 관찰하여 평가하였다. 당-인지질 유도체를 함유한 리포솜은 당-인지질 유도체를 함유하지 않은 리포솜 또는 폴리에틸렌글리콜(PEG) 수식 리포솜에 비해 단백흡착의 양과 혈청 내 입자크기의 변화가 적은 것으로 관찰되었다. 이당이 결합된 DSPE와 이를 함유한 리포솜은 혈액내 안정성이 향상되어 체순환계 내에서 장시간 순환이 가능한 약물전달체로서 유용할 것이라 사료된다. Liposomes having particle size from several tens to hundreds nanometers are efficient carriers for injectable drug delivery. Enhancement of liposome stability in bloodstream has been studied because of its relatively short circulation time and fast clearance from human body by reticuloendothelial system (RES) in blood vessel. In this study, new disaccharide-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) derivatives in which lactose or sucrose as the disaccharide molecule was conjugated covalently to DSPE were synthesized. Liposomes of which surface had disaccharide molecules were prepared by incorporating the disaccharide-DSPE into liposomes as one of their lipid components. Particle size of the prepared liposomes was approximately 100 nm. The liposomes of which surface were modified with the disaccharide-DSPE showed -25 mV of zeta potential value due to the presence of hydroxyl groups on their surface, while the unmodified control liposomes showed -10 mV of zeta potential value. Loading efficiency of model drug, doxorubicin, into liposomes was about 90%. Stability of the disaccharide-modified liposomes in vitro was evaluated by monitoring the amount of protein adsorption and particle size of the liposomes in serum. Disaccharide-modified liposomes were more stable in serum than unmodified control liposomes or polyethyleneglycol (PEG)-modified liposomes due to less adsorption of serum protein and hence less increase of their particle size. The liposomes of which surface was modified with disaccharide-DSPE conjugate can be used as long-circulating carriers for drugs having high toxicity or short half-life time due to their enhanced stability in blood circulatory system.

Hydroxyapatite/Chondoroitin Sulfate Microparticles as Time-Controlled Release Carrier of Proteins

Time-controlled releases of proteins from hydroxyapatite/chondroitin sulfate (HAp/ChS) spherical microparticles were achieved by the addition of zinc cation into the mixture solutions of HAp/ChS and protein as a novel formulation. The initial bursts of proteins, such as cytochrome c and bovine serum albumin, were apparently suppressed by the amount of zinc cation, which could be attributed to the formation of coordinate bonds of zinc cation among proteins and/or ChS moleculars. The increase of molecular lengths of ChS chains decreased the adsorbed amount of proteins, which did not apparently affected to the release of proteins.

Preparation of protein-adsorption-resistant polyethersulfone ultrafiltration membranes through surface segregation of amphiphilic comb copolymer

An amphiphilic comb copolymer with polystyrene as hydrophobic parts and poly(ethyl glycol) (PEG) as hydrophilic parts was blended with polyethersulfone (PES) to fabricate high protein-adsorption-resistant ultrafiltration membranes through a phase inversion process. The hydrophilic PEG segments were spontaneously segregated to the membrane surface during immersion precipitation, which was confirmed by the water contact angle measurement and X-ray photoelectron spectroscopy (XPS). The PEG-enriched surface exhibited higher hydrophilicity and the amount of protein adsorption was remarkably decreased from 6.8 to 0.5 μg/cm 2. Ultrafiltration experiments showed that the antifouling property of the blend membranes was considerably improved with slight change of permeation properties, and the resistance caused by irreversible fouling was remarkably decreased from 2.70 × 10 12 to 0.34 × 10 12 m −1. The superior protein-adsorption-resistance rendered blend membranes more desirable recycling property. The flux recovery ratio was remained as high as 80.4% after three cycles of BSA ultrafiltration.

Cell Adhesion and Proliferation Patterns on Mixed Self-assembled Monolayers Carrying Various Patios of Hydroxyl and Methyl Groups

Mixed self-assembled monolayers (SAMs) with hydroxyl and methyl surface groups were prepared as biomaterial surface models which were well-controlled and well-defined. The surface properties of mixed SAMs were examined by X-ray photoelectron spectroscopy and water contact angle measurement. It was found that the parameter of water contact angle more accurately reflected the surface compositions of mixed SAMs than by the mixing ratio of the two alkanethiols. Cell adhesion and growth were also examined on mixed SAMs of various wettability conditions. It was found that amount of serum protein adsorption changed with the surface composition. To examine the effect of surface composition on cell growth pattern, four cell types--C3H10T1/2-clone 8, L929, UVB6-2.1A, and MC3T3E1--were incubated on mixed SAMs for three or six days. Differences in cell growth pattern against wettability were clearly recognized for each cell type. In light of the results obtained in this study, the relationship between the biocompatibility of biomaterials and surface factors were thus clarified.

Cell Adhesion on Nanofibrous Polytetrafluoroethylene (nPTFE)

Here, we described the in vitro biocompatibility of a novel nanostructured surface composed of PTFE as a potential polymer for the prevention of adverse host reactions to implanted devices. The foreign body response is characterized at the tissue-material interface by several layers of macrophages and large multinucleated cells known as foreign body giant cells (FBGC), and a fibrous capsule. The nanofibers of nanofibrous PTFE (nPTFE) range in size from 20 to 30 nm in width and 3-4 mm in length. Glass surfaces coated with nPTFE (produced by jet-blowing of PTFE 601A) were tested under in vitro conditions to characterize the amount of protein adsorption, cell adhesion, and cell viability. We have shown that nPTFE adsorbs 495 +/- 100 ng of bovine serum albumin (BSA) per cm2. This level was considerably higher than planar PTFE, most likely due to the increase in hydrophobicity and available surface area, both a result of the nanoarchitecture. Endothelial cells and macrophages were used to determine the degree of cell adsorption on the surface of the nanostructured polymer. Both cell types were significantly more round and occupied less area on nPTFE as compared to tissue culture polystyrene (TCPS). Furthermore, a larger majority of the cells on the nPTFE were dead compared to TCPS, at dead-to-live ratios of 778 +/- 271 to 1 and 23 +/- 5.6 to 1, respectively. Since there was a high amount of cell death (due to either apoptosis or necrosis), and the foreign body response is a form of chronic inflammation, an 18 cytokine Luminex panel was performed on the supernatant from macrophages adherent on nPTFE and TCPS. As a positive control for inflammation, lipopolysaccharide (LPS) was added to macrophages on TCPS to estimate the maximum inflammation response of the macrophages. From the data presented with respect to IL-1, TNF-alpha, IFN-gamma, and IL-5, we concluded that nPTFE is nonimmunogenic and should not yield a huge inflammatory response in vivo, and cell death observed on the surface of nPTFE was likely due to apoptosis resulting from the inability of cells to spread on these surface. On the basis of the production of IL-1, IL-6, IL-4, and GM-CSF, we concluded that FBGC formation on nPTFE may be decreased as compared to materials known to elicit FBGC formation in vivo.

Reduction of nonspecific protein binding on surface plasmon resonance biosensors

Reduction of the nonspecific serum protein adsorption on a gold surface to levels low enough to allow the detection of biomarkers in complex media has been achieved using the N-hydroxysuccinimide (NHS) ester of 16-mercaptohexadecanoic acid. Carboxymethylated dextran (CM dextran), which is widely used, nonspecifically adsorbs enough proteins to mask the signal from target biomarkers in complex solutions such as serum or blood. The use of short-chain thiols greatly reduces the amount of nonspecific protein adsorption. Mixed layers of 11-mercaptoundecanoic acid or the NHS ester of 11-mercaptoundecanoic acid mixed layers with either 11-mercaptoundecanol or undecanethiol, and 16-mercaptohexadecanoic acid or the NHS ester of 16-mercaptohexadecanoic acid with hexadecanethiol, were also investigated for nonspecific protein binding properties as well as for biomarker signal response. The NHS ester of 16-mercaptohexadecanoic acid exhibits the largest signal for the biomarker myoglobin (including CM dextran) while offering a significantly diminished amount of nonspecific binding. The sensor has also been shown to detect interleukin-6 in cell culture media containing protein concentrations of at least 4 mg/mL.

The Preparation and Characterisation of a Novel Siloxane Biomaterial

It is well known that polysiloxane interacts strongly with proteins and has been shown to adsorb significant quantities of protein due to its strong hydrophobicity, which often promotes conformational and orientational changes in the adsorbed proteins so as to decrease their biocompatibility. Therefore, the control of protein adsorption to biomaterials and the preparation of suitable siloxane biomaterials become necessary and significant for the actual application. In this paper, a novel siloxane biomaterial (siloxane microspheres) was prepared by dispersion polymerization under kinetically controlled conditions. And then, the siloxane biomaterial was characterized by particle size analyser, X-ray Photoelectron Spectroscopy (XPS) and static contact angle. The siloxane microspheres were used as the matrix, and bovine serum albumin (BSA) as the template protein, the adsorption has been investigated. The study shows that the amount of protein adsorption (BSA) is higher in the prescence of siloxane monomer than than in its absence, and the effects become more evident with the increase in the contents of siloxane monomer. This indicates that its hydrophobicity of siloxane microspheres can be shepherded through the amount of siloxane monomer added.

Generation of anti-biofouling ultrafiltration membrane surface by blending novel branched amphiphilic polymers with polyethersulfone

Branched amphiphilic copolymers with peculiar structural and physicochemical characteristics may find promising application in generating fouling-resistant membrane surface. In this study, novel branched amphiphilic copolymers, P123-b-PEGs, were synthesized by reacting Pluronic P123 with PEG400 using PCl 3 as a conjugation reagent, and this novel amphiphilic copolymers were blended with polyethersulfone (PES) to prepare ultrafiltration membranes with superior protein-adsorption-resistant ability. The static water contact angle and X-ray photoelectron spectroscopy revealed the remarkable enrichment of PEO segments at blend membrane surface, as well as the inherent relationship between near-surface coverage and PEO arm number in P123-b-PEG copolymers. Investigations on protein adsorption on blend membranes showed that the protein adsorption amount was significantly decreased. The results of ultrafiltration experiments revealed that the reversible fouling resistance composed the dominated part of total fouling resistance, which endowed the blend membranes containing P123-b-PEG copolymers with higher flux recovery ratio.

Model salad dressing emulsion stability as affected by the type of the lupin seed protein isolate

AbstractModel salad dressing emulsions of an oil volume fraction of 0.50 were prepared using two types of lupin seed protein isolate (LSPI) differing in the method applied for their isolation and their protein composition. The dressing stability against creaming and droplet coalescence were studied and correlated with data on oil droplet size, rheological characteristics and the amount of protein adsorption at the droplet surfaces. Model salad dressing emulsions containing the isolate, mainly composed of lupin globulins, exhibited higher stability and more pronounced rheological characteristics compared to those prepared with the isolate enriched in albumins or with the mixture of the two isolates. The lupin albumins appeared to displace the globulins from the droplet surfaces, following competitive adsorption from mixtures of the two types of the lupin isolates. The results are discussed in terms of droplet interaction and rearrangement as they are influenced by the presence of the adsorbed protein molecules and aggregates which appear to determine long‐term stability of the emulsion systems. Copyright © 2006 Society of Chemical Industry

Characterization of a non-fouling ultrafiltration membrane

This report describes the properties of surface-modified poly(vinylidene fluoride) (PVDF) membranes. These membranes were created by coating hydrophilic polymers on the support PVDF membrane to reduce the tendency to protein fouling. The modified membranes with different molecular weight cut-off (MWCO) were characterized by filtration studies using bovine serum albumin (BSA) and an enzyme solution as test media, and the membranes exhibited the non-fouling property. The surface chemistry of the unmodified and modified PVDF membranes was characterized by X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectroscopy (ToF-SIMS). These surface sensitive techniques were used to confirm the successful surface modification. ToF-SIMS imaging visualizes the distribution of the coating layer on the PVDF membrane. Furthermore, the amount of protein adsorption onto the membrane surfaces was determined by the radio labeling technique. For the protein adsorption experiments, we have developed a method, which ensures that protein molecules only adsorb on the membrane surface, and that no adsorption on the pore walls of the porous sub-layer takes place.

Self-consistent field theory of protein adsorption in a non-Gaussian polyelectrolyte brush

To describe adsorption of globular protein molecules in a polyelectrolyte brush we use the strong-stretching approximation of the Edwards self-consistent field equation, combined with corrections for a non-Gaussian brush. To describe chemical potentials in this mixture of (globular) species of widely varying sizes (ions, brush polyelectrolyte segments, globular protein molecules), we use the Boublik-Mansoori-Carnahan-Starling-Leland equation of state derived for polydisperse mixtures of spherical particles. The polyelectrolyte chain is described in this approach as a string of beads with the beads of a size related to the chain diameter. We use the one-dimensional Poisson equation to describe the electrostatic field and include the ionizable character of both the brush polyions and the protein molecules. This model explains the experimental observation of high amounts of protein adsorption in a polyacid brush for values above the isoelectric point of the protein as being due to charge reversal of the protein molecules upon entry in the brush. We find a distinct minimum in protein concentration near the edge of the brush. With increasing this barrier to protein transfer becomes larger, but much less so when we increase the ionic strength, a difference that might relate to an experimentally observed difference in the protein release rate in these two cases. A free energy analysis shows that the release of small ions from the brush and the increase of brush ionization are the two driving forces for protein adsorption in a like-charged brush.

Remarkable Reduction of Irreversible Fouling and Improvement of the Permeation Properties of Poly(ether sulfone) Ultrafiltration Membranes by Blending with Pluronic F127

Hydrophilic modification of ultrafiltration membranes was achieved through blending of Pluronic F127 with poly(ether sulfone) (PES). The chemical composition and morphology changes of the membrane surface were confirmed by water contact angle, X-ray photoelectron spectroscopy, scanning electron microscopy, and protein adsorption measurements. The decreased static water contact angle with an increase in the Pluronic F127 content indicated an increase of surface hydrophilicity. XPS analysis revealed enrichment of PEO segments of Pluronic F127 at the membrane surface. The apparent protein adsorption amount decreased significantly from 56.2 to 0 microg/cm(2) when the Pluronic F127 content varied from 0% to 10.5%, which indicated that the blend membrane had an excellent ability to resist protein adsorption. The ultrafiltration experiments revealed that the Pluronic F127 content had little influence on the protein rejection ratio and pure water flux. Most importantly, at a high Pluronic F127 content membrane fouling, especially irreversible fouling, has been remarkably reduced. The flux recoveries of blend membranes reached as high as 90% after periodic cleaning in three cycles.

Change of ζ Potential of Biocompatible Colloidal Oxide Particles upon Adsorption of Bovine Serum Albumin and Lysozyme

The amounts of negatively charged bovine serum albumin and positively charged lysozyme adsorbed on alumina, silica, titania, and zirconia particles (diameters 73 to 271 nm) in aqueous suspensions are measured. The adsorbed proteins change the zeta potentials and the isoelectric points (IEP) of the oxide particles. The added to adsorbed protein ratios at pH 7.5 are compared with the protein treated particle zeta potentials. It is found that the amounts of adsorbed proteins on the alumina, silica, and titania (but not on the zirconia) particle surfaces are highly correlated with the zeta potential. For the slightly less hydrophilic zirconia particles high amounts of protein adsorption are observed even under repulsive electrostatic conditions. One reason could be that the hydrophobic effect plays a more important role for zirconia than electrostatic interaction.

Modification of polysulfone ultrafiltration membranes by CO 2 plasma treatment

Carbon dioxide plasmas were used to modify hydrophobic polysulfone ultrafiltration membranes to create hydrophilic surfaces throughout the membrane structure. The water contact angle of the upstream side of the membrane (facing the plasma) decreased to zero after treatment and did not change even after several months of aging. The water contact angle of the downstream side decreased with increasing CO 2 plasma treatment time and became zero for treatment times ≥ 1 min ( P = 10 W). Functional groups introduced by CO 2 plasma treatment were examined using X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). For the treated membranes, the atomic concentration of oxygen increased dramatically and small amounts of nitrogen incorporation were also observed. Membrane performance was tested with water flux measurements as well as protein fouling studies. For treated membranes, the water flux recovery measured after protein fouling was significantly higher than that for control membranes, with nearly 100% recovery after gentle cleaning in water. Moreover, the amount of protein adsorption decreased by over 75% for the treated membranes compared to control membranes. This suggests the protein fouling layer is essentially completely reversible on the CO 2 plasma treated membranes.

How polysulfone dialysis membranes containing polyvinylpyrrolidone achieve excellent biocompatibility?

Polysulfone (PS) dialysis membranes hydrophilized by blending polyvinylpyrrolidone (PVP) are well known to have excellent biocompatibility in clinical use. The objective of the present study is thus to clarify how PVP improves biocompatibility of PS membranes and furthermore to develop a patient-friendly PS dialysis membrane with higher biocompatibility. Biocompatibility based on both lactate dehydrogenase (LDH) activity and amount of protein adsorption was greatly different among four commercially available PS hollow-fiber dialysis membranes. PVP present on the inner surface of the hollow fiber was quantitatively determined by X-ray photoelectron spectroscopy (XPS), demonstrating the amount of PVP to be varying for each membrane. Structure parameters such as surface roughness, three-dimensional surface area and polymer particle diameter, indications of the physicochemical properties of the membranes, were measured on the observed inner surface images in both wet and dry conditions by atomic force microscopy (AFM) to account for dependence of biocompatibility on these structure parameters. The higher regularity polymer particle structure has in the wet condition, the lower wet/dry ratio surface roughness has and the larger wet/dry ratio polymer particle diameter has, that is, the more greatly the polymer particles swell by wetting, the higher biocompatibility is achieved by “cushion effect”.

Characterization of poly(L-lysine)-graft-poly(ethylene glycol) assembled monolayers on niobium pentoxide substrates using time-of-flight secondary ion mass spectrometry and multivariate analysis.

Control of protein adsorption onto solid surfaces is a critical area of biomaterials and biosensors research. Application of high performance surface analysis techniques to these problems can improve the rational design and understanding of coatings that control protein adsorption. We have used static time-of-flight secondary ion mass spectrometry (TOF-SIMS) to investigate several poly(L-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) adlayers adsorbed electrostatically onto negatively charged niobium pentoxide (Nb(2)O(5)) substrates. By varying the PEG graft ratio (i.e., the number of lysine monomers per grafted PEG chain) and the molecular weights of the PLL and PEG polymers, the amount of protein adsorption can be tailored between 1 and 300 ng/cm(2). Detailed multivariate analysis using principal component analysis (PCA) of the positive and negative ion TOF-SIMS spectra showed changes in the outermost surface of the polymer films that were related to the density and molecular weight of the PEG chains on the surface. However, no significant differences were noted due to PLL molecular weight, despite observed differences in the serum adsorption characteristics for adlayers of PLL-g-PEG polymers with different PLL molecular weights. From the PCA results, multivariate peak intensity ratios were developed that correlated with the thickness of the adlayer and the enrichment of the PEG chains and the methoxy terminus of the PEG chains at the outermost surface of the adlayer. Furthermore, partial least squares regression was used to correlate the TOF-SIMS spectra with the amount of protein adsorption, resulting in a predictive model for determining the amount of protein adsorption on the basis of the TOF-SIMS spectra. The accuracy of the prediction of the amount of serum adsorption depended on the molecular weight of the PLL and PEG polymers and the PEG graft ratio. The combination of multivariate analysis and static TOF-SIMS provides detailed information on the surface chemistry and insight into the mechanism for protein resistance of the coatings.

Protein adsorption to planar electrochemical sensors and sensor materials

Abstract In electrochemical sensing devices, aimed for acute and chronic in vivo application, the active surface of the sensor is often negligible compared to the overall surface area of the device in contact with the biological host. Consequently, to minimize the perturbation of an implanted sensor on the in vivo environment the chemical composition and surface texturing of the complete device (the active sensor, sensor substrate, and “accessories”) have to be considered. In our work, the adsorption of three abundant proteins (albumin, IgG, and fibrinogen) was determined quantitatively on untreated and modified sensor substrates and sensing membrane surfaces. In this study, a flexible polyimide-based material (Kapton ®) was used as sensor substrate with or without an amorphous diamond-like carbon (DLC) or an amorphous oxygen-containing DLC (o-DLC) coating. The ion-sensitive membranes were cast from high-molecular-weight (HMW) or carboxylated poly(vinyl chloride) (PVC) and were doped with increasing concentrations of highly hydrophilic poly(ethylene oxide) (PEO). The potentiometric characteristics of the potassium-selective membranes cast with up to 6 % PEO were the same as those without PEO. However, the PEO-modified PVC membranes elicited a large amount of protein adsorption, especially in terms of albumin.

Protein and bacterial fouling characteristics of peptide and antibody decorated surfaces of PEG-poly(acrylic acid) co-polymers

The potential for base poly(ethylene glycol) graft poly(acrylic acid) PEG-g-PA copolymers and surface-modified PEG-g-PA materials to inhibit random protein fouling and bacterial adhesion are investigated. PEG-g-PA co-polymers were synthesized that inhibited non-specific protein and cellular adhesion. PEG-g-PA co-polymers were then covalently modified with either cell adhesion peptides (YRGDS, YEILDV) or fragments of antibodies to monocyte/macrophage integrin receptors (Anti-VLA4, Anti- β 1, Anti- β 2, and Anti-CD64) known to enhance macrophage adhesion and, perhaps, modulate their activation. Materials produced in this work were characterized using: hydrophobicity by contact angle; angle-resolved X-ray Photoelectron Spectroscopy to confirm the presence of PEG in the bulk material and the surface; degree of hydration; differential scanning calorimetry; and thermal gravimetric analysis. To evaluate the non-fouling efficacy of the various modified surfaces, three proteins, human serum albumin, human fibronectin (Fraction I) and human immunoglobulin were 125I labeled. Samples of base PEG-g-PA and PEG-g-PA, modified with various peptides, were exposed to solutions containing either 2 or 200 μg/ml of one of the labeled proteins at 37°C for 24 h. PEG-g-PA substrata modified with directly bound peptides exhibited protein adsorption that varied depending upon the surface bounded peptide. PEG-g-PA modified with peptides linked by linear PEG tethers reduced protein adsorption at 24 h by ∼45% in comparison to PEG-g-PA. Peptides linked by way of StarPEO and StarlikePEO tethers further decreased protein adsorption in comparison to PEG-g-PA. The ability of peptide:PEOtethers to inhibit protein adsorption appeared to be a function of type and surface coverage of the PEO tether and not influenced by the amount or molecular structure the tethered peptide. Peptides directly coupled to the PEG-g-PA increased the amount of protein fouling relative to controls and there appeared to be some dependency of the amount of protein adsorption on which peptide was tethered. Two 14C-labeled pathogens, Staphylococcus epidermidis and Pseudomonas aeruginosa, were used to quantify the degree of bacterial adhesion using two types of laminar flow cell chambers; one that provided invasive sampling of the target substrata and one that provided non-invasive microscopic surveillance of adhering bacterial cells. Attachment of both species to PEG-g-PA and peptide-modified PEG-g-PA was reduced compared to the basic poly(acrylic acid). Presence of peptides on the surface, whether directly bound or bound by the PEO tether did not influence adhesion of P. aeruginosa relative to controls. S. epidermidis adhesion rates increased slightly for those materials where peptides were directly bound to the surface but were reduced relative to base PEG-g-PA when peptides were bound by PEO tethers. All PEG-g-PA surfaces modified with fragments of monoclonal antibodies dramatically enhanced bacterial initial adhesion rates and maximum extent of attachment.

Effects of heat on physicochemical properties of whey protein-stabilised emulsions

The effect of heating has been studied for whey protein-stabilised oil-in-water emulsions (25.0% (w/w) soybean oil, 3.0% (w/w) whey protein isolate, pH 7.0). These emulsions were heated between 55 and 95 °C as a function of time and the effect on particle size distribution, adsorbed protein amount, protein conformation and rheological properties was determined. Heating the emulsions as a function of temperature for 25 min resulted in an increase of the mean diameter ( d 32) and shear viscosity with a maximum at 75 °C. Heating of the emulsions at different temperatures as a function of time in all cases resulted in a curve with a maximum for d 32. A maximum increase of d 32 was observed after about 45 min at 75 °C and after 6–8 min at 90 °C. Similar trends were observed with viscosity measurements. Confocal scanning laser micrographs showed that after 8 min of heating at 90 °C large, loose aggregates of oil droplets were formed, while after 20 min of heating compact aggregates of two or three emulsion droplets remained. An increase of the adsorbed amount of protein was found with increasing heating temperature. Plateau values were reached after 10 min of heating at 75 °C and after 5 min of heating at 90 °C. Based on these results we concluded that in the whole process of aggregation of whey protein-stabilised emulsions an essential role is played by the non-adsorbed protein fraction, that the kinetics of the aggregation of whey protein-stabilised emulsions follow similar trends as those for heated whey protein solutions and that upon prolonged heating rearrangements take place leading to deaggregation of initially formed large, loose aggregates of emulsion droplets into smaller, more compact ones.