Adsorption of IgG on/in a PAH/PSS multilayer film: Layer structure and cell response

The binding of immunogloblulins (IgG) (mouse monoclonal recognizing IFNγ) on precoated polystyrene or silica surfaces by the layer-by-layer technique has been investigated with QCM-D and DPI. The aim of the work was to increase the sensitivity of the conventional enzyme-linked immunosorbent spot (ELISpot) assay. The polyelectrolytes used to build the multilayers were poly(allylamine hydrochloride) (PAH)/poly(sodium 4-styrenesulfonate) (PSS) alternately adsorbed from 150mM NaCl. The multilayer build up is linear and the internal structure of the PAH/PSS multilayer is compact and rigid as observed by low relative water content (20-25%) and high layer refractive index (n～1.5) after the formation of five bilayers. Incorporation of IgG within the PAH/PSS multilayer did not give rise to overcharging and did not affect the linear build up. ELISpot test on PAH/PSS multilayer modified polystyrene wells showed that the cytokine response was significantly smaller than on the regular PVDF backed polystyrene wells. This may be due to the compact and rigid nature of the PAH/PSS multilayer, which does not allow formation of the kind of three dimensional support needed to achieve bioactive IgG binding to the surface. Immunological tests of the polyelectrolyte multilayers in the absence of IgG showed that PSS terminated PAH/PSS multilayer did not induce any cytokine response whereas PAH terminated did, which suggests that PSS totally covers the surface from the cells point of view.     

Study of film structure and adsorption kinetics of polyelectrolyte multilayer films: effect of pH and polymer concentration

The alternate adsorption of polycation poly(allylamine hydrochloride)(PAH) and the sodium salt of the polymeric dye poly(1-[ p-(3'-carboxy-4'-hydroxyphenylazo)benzenesulfonamido]-1,2-ethandiyl)(PCBS) on quartz crystals coated with silica was studied to understand the structural properties and adsorption kinetics of these films using a combination of quartz crystal microbalance with dissipation monitoring (QCM-D), absorbance, and ellipsometry measurements. In-situ deposition of the polycation PAH on QCM crystals was monitored, followed by rinsing with water and then deposition of the polyanion PCBS. The effects of polymer concentration and pH on film structure, composition and adsorption kinetics were probed. The polymers were adsorbed at neutral pH conditions and at elevated pH conditions where PAH was essentially uncharged to obtain much thicker films. The change in the resonant frequency, Deltaf, of the QCM-D showed a linear decrease with the number of bilayers, a finding consistent with absorbance and ellipsometric thickness measurements which showed linear growth of film thickness. By using the Delta f ratios of PCBS to PAH, the molar ratios of repeat units of PCBS to PAH in the bilayer films as determined by QCM-D were approximately 1:1 at polyelectrolyte concentrations 5-10 mM repeat unit, indicating complete dissociation of the ionic groups. The frequency and dissipation data from the QCM-D experiments were analyzed with the Voigt model to estimate the thickness of the hydrated films which were then compared with thicknesses of dry films measured by ellipsometry. This led to estimates of the water content of the films to be approximately 45 wt %. In addition to the QCM-D, some films were also characterized by a QCM which measures only the first harmonic without dissipation monitoring. For the deposition conditions studied, the deposited mass values measured by the QCM's first harmonic were similar to the results obtained using higher harmonics from QCM-D, indicating that the self-assembled polyelectrolyte films were rigid.     

Layer-by-layer assemblies of chitosan and heparin: effect of solution ionic strength and pH

The growth of polysaccharide multilayers consisting of positively charged chitosan (CH) and negatively charged heparin (HEP) was monitored in situ by employing a quartz crystal microbalance (QCM-D) and dual-polarization interferometry (DPI). The main focus was on how the physicochemical properties of the solution affect the growth and structure of the resulting multilayer film. These results showed that when increasing the ionic strength of the polysaccharide solutions at a fixed pH, both the "dry" (optical) (DPI) mass and wet (QCM) mass of the adsorbed multilayer film increased. The same effect was found when increasing the pH while keeping the ionic strength constant. Furthermore, the growth of multilayers showed an exponential-like behavior independent of the solution conditions that were used in this study. It was also established that chitosan was the predominant species present in the chitosan-heparin multilayer film. We discuss the viscoelastic properties of the adsorbed layers and their variation during the multilayer buildup. Interestingly and contrary to common interpretation of the QCM-D results, we found that under one particular solution condition (pH 4.2 and 30 mM NaCl) the increase in the dissipation of oscillation energy from the adsorbed layer was a consequence of layer stiffening rather than indicating a more hydrated and viscous film. On the basis of the widely used Voigt viscoelastic model for an adsorbed layer, we show that it is the film viscosity and shear that define the layer viscoelasticity (structure) of the film and not the absolute value of energy dissipation, which in fact can be very misleading.     

Visco-elastic and adhesive properties of adsorbed polyelectrolyte multilayers determined in situ with QCM-D and AFM measurements

The build-up of multilayers constructed from polyallylamine hydrochloride (PAH) and polyacrylic acid (PAA) under different pH conditions was continuously monitored using the quartz crystal microbalance with dissipation. The adsorbed amount of polymer as well as the amount of coupled water was determined. Furthermore, from dissipation measurements, it was possible to determine the visco-elastic properties of the adsorbed multilayer. These properties were highly dependent on the polyelectrolyte present in the outermost layer. The multilayer was far more rigid and elastic with PAA as the outermost layer. Furthermore, a link has been established between the conformability or rigidity of a multilayer covered surface and the adhesion between such surfaces. Adhesion measurements using the atomic force microscope showed a greater pull-off force when the more viscous PAH was present in the outermost layer.     

Characteristics of polyelectrolyte multilayers: effect of PEI anchoring layer and posttreatment after deposition

The influence of a first (anchoring) layer and film treatment on the structure and properties of polyelectrolyte multilayer (PEM) films obtained from polyallylamine hydrochloride (PAH) and polysodium 4-styrenesulfonate (PSS) was studied. Branched polyethyleneimine (PEI) was used as an anchoring layer. The film thickness was measured by ellipsometry. Complementary X-ray reflectometry and AFM experiments were performed to study the change in the interfacial roughness. We found that the thickness of the PEM films increased linearly with the number of layers and depended on the presence of an anchoring PEI layer. Thicker films were obtained for multilayers having PEI as the first layer comparing to films having the same number of layers but consisting of PAH/PSS only. We investigated the wettability of PEM surfaces using direct image analysis of the shape of sessile water drops. Periodic oscillations in contact angle were observed. PAH-terminated films were more hydrophobic than films with PSS as the outermost layer. The effect of long time conditioning of PEM films in solutions of various pH's or salt (NaCl) concentrations was also examined. Salt or base solutions induced modification in wetting properties of the polyelectrolyte multilayers but had a negligible effect on the film thickness.     

Surface interactions during polyelectrolyte multilayer build-up. 2. The effect of ionic strength on the structure of preformed multilayers

Interactions between surfaces bearing multilayer films of poly(allylamine hydrochloride) (PAH) and poly(styrenesulfonate sodium salt) (PSS) were investigated across a range of aqueous KBr solutions. Three layer films (PAH/PSS/PAH) were preassembled on mica surfaces, and the resulting interactions were measured with the interferometric surface force apparatus (SFA). Increasing the ionic strength of the medium resulted in a progressive swelling of the multilayer films. Interactions in solutions containing more than 10(-3) M KBr were dominated by a long-ranged steric repulsion originating from compression of polyelectrolyte segments extending into solution. In 10(-1) M KBr, repeated measurements at the same contact position showed a considerable reduction of the range and the strength of the steric force, indicating a flattening of the film during initial approach. Furthermore, this flattening was irreversible on the time scale of the experiments, and measurements performed up to 72 h after the initial compression showed no signs of relaxation. These studies aid in understanding the dominant interactions between polyelectrolyte multilayers, including polyelectrolyte films deposited on colloidal particles, which is important for the preparation of colloidally stable nanoengineered particles.     

Ion distribution in polyelectrolyte multilayers with standing-wave X-ray fluorescence

Absolute ion concentration and its profile across polyelectrolyte multilayer films were studied. The films were prepared by alternating adsorption of polyanions and polycations from aqueous solution. Standing-wave X-ray fluorescence was used to map the ion profile. The well-studied multilayer system PSS/PAH was investigated, and bromide ions were used as probe entities. The results show that the sign of the charge of the outermost layer and the washing procedure after finishing the preparation have a decisive effect on the ion concentration and the ion profile. Multilayers with PSS as the outermost layer contain fewer bromide ions than the PAH-terminated multilayers. Exposure to water washes the ions out, but even after 6 h of washing, not all of the bromide ions had been removed.     

Influence of solvent quality on the growth of polyelectrolyte multilayers

The effect of solvent conditions on the growth of polyelectrolyte (PE) multilayer films comprising poly(allylamine hydrochloride) (PAH) and poly(styrenesulfonate sodium salt) (PSS) on planar substrates was investigated by means of surface plasmon resonance spectroscopy (SPRS), quartz crystal microbalance (QCM), and atomic force microscopy techniques. The solvent quality was varied by the addition of ethanol to the PE solutions used for deposition of the layers, thus tuning the relative strength of electrostatic and secondary intermolecular and intramolecular interactions. Experiments were performed with PE solutions both without added electrolyte and containing 0.5 M NaCl. Decreasing the solvent quality (i.e., increasing the amount of ethanol in the adsorption solution) resulted in a marked increase of both the multilayer film thickness and mass loading, as determined from the SPRS spectra and QCM frequency shifts, respectively. With the solution composition approaching the precipitation point, thick PAH/PSS films were formed due to the screening of the electrostatic intra- and interchain repulsions and enhanced hydrophobic interactions between the polyelectrolyte chains. However, the films formed from water/ethanol mixtures remained stable upon subsequent exposure to water or salt-containing solutions: no significant film desorption occurred after up to 24 h of exposure to water or 0.5 M NaCl solutions. In addition, the effect of postdeposition exposure to water/ethanol mixtures was investigated for PE multilayers assembled from aqueous solutions. In this case, the optical thickness of the films was determined during exposure to water/ethanol mixtures, and instead of swelling, the polyelectrolyte films collapse to the surface as a result of the unfavorable segment-solvent interactions.     

Layer-by-layer electrostatic self-assembly of single-wall carbon nanotube polyelectrolytes

We have used anionic and cationic single-wall carbon nanotube polyelectrolytes (SWNT-PEs), prepared by the noncovalent adsorption of ionic naphthalene or pyrene derivatives on nanotube sidewalls, for the layer-by-layer self-assembly to prepare multilayers from carbon nanotubes with polycations, such as poly(diallyldimethylammonium) or poly(allylamine hydrochloride) (PDADMA or PAH, respectively), and polyanions (poly(styrenesulfonate), PSS). This is a general and powerful technique for the fabrication of thin carbon nanotube films of arbitrary composition and architecture and allows also an easy preparation of all-SWNT (SWNT/SWNT) multilayers. The multilayers were characterized with vis-near-IR spectroscopy, X-ray photoelectron spectroscopy (XPS), surface plasmon resonance (SPR) measurements, atomic force microscopy (AFM), and imaging ellipsometry. The charge compensation in multilayers is mainly intrinsic, which shows the electrostatic nature of the self-assembly process. The multilayer growth is linear after the initial layers, and in SWNT/polyelectrolyte films it can be greatly accelerated by increasing the ionic strength in the SWNT solution. However, SWNT/SWNT multilayers are much more inert to the effect of added electrolyte. In SWNT/SWNT multilayers, the adsorption results in the deposition of 1-3 theoretical nanotube monolayers per adsorbed layer, whereas the nominal SWNT layer thickness is 2-3 times higher in SWNT/polyelectrolyte films prepared with added electrolyte. AFM images show that the multilayers contain a random network of nanotube bundles lying on the surface. Flexible polyelectrolytes (e.g., PDADMA, PSS) probably surround the nanotubes and bind them together. On macroscopic scale, the surface roughness of the multilayers depends on the components and increases with the film thickness.     

Assembly of poly(N-isopropylacrylamide)-co-acrylic acid microgel thin films on polyelectrolyte multilayers: Effects of polyelectrolyte layer thickness, surface charge, and microgel solution pH

Temperature- and pH-sensitive poly(N-isopropylacrylamide)?Cco-acrylic acid (pNIPAm-co-AAc) microgels were deposited on glass substrates coated with polyelectrolyte multilayers composed of the polycation poly(allylamine hydrochloride) (PAH) and the polyanion poly(sodium 4-styrenesulfonate) (PSS). The microgel density and structure of the resultant films were investigated as a function of: (1) the number of PAH/PSS layers (layer thickness); (2) the charge on the outer layer of the polyelectrolyte multilayer film; and (3) the pH of microgel deposition solution. The resultant films were studied by differential interference contrast optical microscopy, atomic force microscopy, and scanning electron microscopy. It was found that the coverage of the microgels on the surface was a complex function of the pH of the deposition solution, the charge on the outer layer of the polyelectrolyte thin film and the PAH/PSS layer thickness; although it appears that microgel charge plays the biggest role in determining the resultant surface coverage.     

Tuning the architecture of cellulose nanocrystal-poly(allylamine hydrochloride) multilayered thin films: influence of dipping parameters

Multilayered thin films consisting of alternating cationic polyelectrolyte, poly(allylamine hydrochloride) (PAH), and anionic cellulose nanocrystals (CNs) were constructed using the dipping procedure by screening different experimental parameters: the drying step between each layer adsorption, the dipping time, the ionic strength of the PAH solution, and the concentration of CNs dispersion. We showed that the drying process and the ionic strength of PAH solution were crucial parameters for the successful construction of 8-bilayer films. Film thickness is mainly influenced by dipping time and CN concentration when using the dipping procedure without drying. Two architectures of adsorbed CN layers-a single or a double layer of CNs-were revealed on the basis of the thickness increment per bilayer, depending on experimental conditions. The layer adsorption process was investigated in real-time using quartz crystal microbalance with dissipation (QCM-D) experiments in an aqueous environment or by incorporating a drying step. On the basis of in situ construction of PAH-CN films in wet media, QCM-D data were indicative of highly hydrated films for which the progressive layer stacking is disturbed or prevented. QCM-D monitoring of CNs and PAH layer adsorption was monitored by incorporating a drying process. The impact of experimental parameters on PAH-CN multilayered construction and on CN layer configuration is discussed. This study offers new opportunities for tailoring the architecture of CN-based multilayer films.     

Role of chain interpenetration in layer-by-layer deposition of polyelectrolytes

The effects of temperature, pH, and salt concentration on the layer-by-layer (LBL) deposition of sodium poly(styrene sulfonate) (PSS)/poly[2-(dimethylamino)ethyl methacrylate] (PDEM) were investigated by use of a quartz crystal microbalance with dissipation (QCM-D). At pH 4, the frequency change (Deltaf) gradually decreased to a constant, indicating that the polyelectrolyte complexes of the layer were not dissolved. As the layer number increased, the -Deltaf oscillatedly increased, indicating that the thickness of the multilayer increased. At the same time, the dissipation change (DeltaD) oscillatedly increased with the layer number, indicating the chain interpenetration or complexation that led to the alternative swelling-and-shrinking of the outermost layer. For the same layer number, as the temperature increased, the amplitude of DeltaD increased, indicating that the chain interpenetration increased. The thickness also increased with temperature. Further increasing the pH to 7 led to a thicker layer, reflected in the larger amplitude of DeltaD. At pH 10, the polyelectrolytes no longer formed multilayers on the surface because of the lack of electrostatic interactions. On the other hand, the addition of NaCl also led to a thickness increase. The amplitude in DeltaD increased with NaCl concentration, indicating that the chain interpenetration increased. Our experiments indicated that the LBL deposition of polyelectrolytes was dominated by the chain interpenetration. Also, the polyelectrolyte complexes in the layer can redissolve into solution from the surface at a high temperature or a high salt concentration.     

Buildup of polyelectrolyte multilayers of polyethyleneimine and microfibrillated cellulose studied by in situ dual-polarization interferometry and quartz crystal microbalance with dissipation

Polyethyleneimine (PEI) and Microfibrillated cellulose (MFC) have been used to buildup polyelectrolyte multilayers (PEM) on silicone oxide and silicone oxynitride surfaces at different pH values and with different electrolyte and polyelectrolyte/colloid concentrations of the components. Consecutive adsorption on these surfaces was studied by in situ dual-polarization interferometry (DPI) and quartz crystal microbalance measurements. The adsorption data obtained from both the techniques showed a steady buildup of multilayers. High pH and electrolyte concentration of the PEI solution was found to be beneficial for achieving a high adsorbed amount of PEI, and hence of MFC, during the buildup of the multilayer. On the other hand, an increase in the electrolyte concentration of the MFC dispersion was found to inhibit the adsorption of MFC onto PEI. The adsorbed amount of MFC was independent of the bulk MFC concentration in the investigated concentration range (15-250 mg/L). Atomic force microscopy measurements were used to image a MFC-treated silicone oxynitride chip from DPI measurements. The surface was found to be almost fully covered by randomly oriented microfibrils after the adsorption of only one bilayer of PEI/MFC. The surface roughness expressed as the rms-roughness over 1 microm2 was calculated to be 4.6 nm (1 bilayer). The adsorbed amount of PEI and MFC and the amount of water entrapped by the individual layers in the multilayer structures were estimated by combining results from the two analytical techniques using the de Feijter formula. These results indicate a total water content of ca. 41% in the PEM.     

双网络增强自支撑多层膜的制备

通过石英晶体微天平(QCM)研究不同最外层对单体、交联剂、引发剂扩散入多层膜的结果显示,聚乙烯亚胺( PEI)为最外层的多层膜更有利于负电荷分子扩散.将(聚乙烯亚胺/聚丙烯酸)12-聚乙烯亚胺[( PEI/PAA)12-PEI]多层膜浸入带有单体、引发剂、交联剂的溶液后,经过光聚合交联与热交联后可形成自支撑膜.XPS数...     

Unusual collapse of highly hydrated polyelectrolyte multilayers with the ionic strength

Highly hydrated polyelectrolyte multilayers (PEMs) were fabricated by “layer by layer” (LBL) assembly of poly (diallyl dimethyl ammonium chloride) (PDADMAC) and poly (sodium 4‐styrene sulfonate) (PSS) in 0.5 M NaCl. Both thickness and hydration of the film were determined in situ as the multilayer was assembled by means of the quartz crystal microbalance with dissipation (QCM‐D) and the Spectroscopic Ellipsometry techniques combined in a single device. For PEMs of 17 total layers in water, a final thickness of up to 300 nm and a hydration of 69% were measured. The response towards the ionic strength was then studied by means of QCM‐D. PEMs of 17 layers, with PDADMAC as last layer, shrunk dramatically and lost water when exposed to aqueous NaCl solutions of increasing concentration. Indeed, a thickness variation up to 100 nm and reduction in the 50% of the water content were observed when the PEM was exposed to 1 M NaCl. On the contrary, PEMs where PSS appears on top showed no measurable change upon the variation in the ionic strength. This brings the possibility to control the responsive character of the PEMs simply by selecting the last polyelectrolyte layer (PDADMAC or PSS) deposited. © 2011 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2011     

Multivalent ion/polyelectrolyte exchange processes in exponentially growing multilayers

We show, in this paper that multivalent ferrocyanide anions can penetrate into exponentially growing (PGA/PAH)n multilayer films whatever the nature of the last deposited layer. These ions are not able to diffuse out of the film when it is brought in contact with a pure buffer solution. However, the contact of this film with a poly(allylamine) (PAH) or a poly(L-glutamic acid) (PGA) solution leads to the release of ferrocyanide ions from the multilayer. It is shown that the release of ferrocyanide anions, when the film is in contact with a PGA solution, is due to the diffusion of the PGA chains into the film so that an exchange between ferrocyanide ions and PGA chains takes place inside the film. On the other hand, PAH chains do not diffuse into PGA/PAH multilayers. When the film is then brought in contact with a PAH solution, the PAH chains from the solution are expected to strongly interact with the ferrocyanide ions and thus induce a diffusion mechanism of the multivalent anions out of the film, the film/solution interface playing the role of a sink for these ions. This work thus shows that interactions between multivalent ions and exponentially growing films are much more complex than expected at first sight and that polyelectrolyte multilayers must be seen as dynamic entities in which diffusion and exchange processes can take place.     

Assembly and mechanical properties of phosphorus dendrimer/polyelectrolyte multilayer microcapsules

We report the preparation, characterization, and mechanical properties of polyelectrolyte/phosphorus dendrimer multilayer microcapsules. The shells of these microcapsules are composed either by alternating poly(styrenesulfonate) (PSS) and positively charged dendrimer G4(NH+Et2Cl-)96 or by alternating poly(allylamine hydrochloride) (PAH) and negatively charged dendrimer G4(CH-COO-Na+)96. The same multilayers were constructed on planar support to examine their layer-by-layer growth and to measure the multilayer thickness. Surface plasmon resonance spectroscopy (SPR) showed regular linear growth of the assembly upon each bilayer deposited. We probe the mechanical properties of these polyelectrolyte/dendrimer microcapsules by measuring force-deformation curves with the atomic force microscope (AFM). The experiment suggests that they are much softer than PSS/PAH microcapsules studied before. This softening is attributed to an enhanced permeability of the polyelectrolyte/dendrimer multilayer shells as compared with multilayers formed by linear polyelectrolytes. In contrast, Young's modulus of both dendrimer-based multilayers was found to be on the same order as that of PSS/PAH multilayers.     

Surface interactions during polyelectrolyte multilayer buildup. 1. Interactions and layer structure in dilute electrolyte solutions

We report the investigation of surface forces between polyelectrolyte multilayers of poly(allylamine hydrochloride) (PAH) and poly(styrenesulfonate sodium salt) (PSS) assembled on mica surfaces during film buildup using a surface force apparatus. Up to four polyelectrolyte layers were prepared on each surface ex situ, and the surface interactions were measured in 10(-4) M KBr solutions. The film thickness under high compressive loads (above 2000 microN/m) increased linearly with the number of deposited layers. In all cases, the interaction between identical surfaces at large separations (>100 A from contact) was dominated by electrostatic double-layer repulsion. By fitting DLVO theory to the experimental force curves, the apparent double-layer potential of the interacting surfaces was calculated. At shorter separations, an additional non-DLVO repulsion was present due to polyelectrolyte chains extending some distance from the surface into solution, thus generating an electrosteric type of repulsion. Forces between dissimilar multilayers (i.e., one of the multilayers terminated with PSS and the other with PAH) were attractive at large separations (30-400 A) owing to a combination of electrostatic attraction and polyelectrolyte bridging.     

Counterions and water in polyelectrolyte multilayers: a tale of two polycations

Attenuated total internal reflectance Fourier transform infrared, ATR-FTIR, spectroscopy was used to compare the water uptake and doping within polyelectrolyte multilayers made from poly(styrene sulfonate), PSS, and a polycation, either poly(allylamine hydrochloride), PAH, or poly(diallyldimethylammonium chloride), PDADMAC. Unlike PDADMA/PSS multilayers, whose water content depended on the solution ionic strength, PAH/PSS multilayers were resistant to doping by NaCl to a concentration of 1.2 M. Using (infrared active) perchlorate salt, the fraction of residual counterions in PDADMA/PSS and PAH/PSS was determined to be 3% and 6%, respectively. The free energy of association between the polymer segments, in the presence of NaClO4, was about 5 kJ mol-1 and -10 kJ mol-1, respectively, for PDADMA/PSS and PAH/PSS, indicating the relatively strong association between the polymer segments in the latter relative to the former. Varying the pH of the solution in contact with the PAH/PSS multilayer revealed a transition to a highly swollen state, interpreted to signal protonation of PAH under much more basic conditions than the pKa of the solution polymer. The increase in the multilayer pKa suggested an interaction energy for PAH/PSS in NaCl of ca. 16 kJ mol-1.     

The electroactive multilayer films of polyelectrolytes and Prussian blue nanoparticles and their application for H2O2 sensors

Prussian blue (PB) nanoparticles were immobilized in polyelectrolyte (PE) multilayers of various compositions and thickness. Films containing nanoparticles and poly(allylamine hydrochloride) (PAH) were formed using the layer-by-layer adsorption method. A layer of branched poly(ethyleneimine) (PEI) was used to anchor the multilayer structure at the surface of a gold electrode. The films exhibited electroactive properties, increasing with the number of deposited PB layers. The properties of PEI/(PB/PAH) _n multilayers were then compared with the ones containing additionally the conductive polymer poly(3,4-ethylenedioxythiophene)–poly(styrenesulfonate) (PEDOT:PSS). We found that the addition of the conductive, water-soluble polymer enhances the electroactive properties of the multilayer films. It also increased sensitivity of the multilayer-covered electrodes for electrochemical detection of hydrogen peroxide.