Glycol Monolayers Research Articles

Tailoring the Surface Chemistry of PEDOT:PSS to Promote Supported Lipid Bilayer Formation

AbstractThis communication reports on a versatile and substrate‐agnostic method to tune the surface chemistry of conducting polymers with the aim of bridging the chemical mismatch between bioelectronic devices and biological systems. As a proof of concept, the surface of poly(3,4‐ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) is grafted with a short‐chain oligoethylene glycol monolayer to favor the formation of cell‐derived supported lipid bilayers (SLBs). This method is tuned to optimize the affinity between the supported lipid bilayer and the conducting polymer, leading to significant improvements in bilayer quality and therefore electronic readouts. To validate the impact of surface functionalization on the system's ability to transduce biological phenomena into quantifiable electronic signals, the activity of a virus commonly used as a surrogate for SARS‐CoV‐2 (mouse hepatitis virus) is monitored with and without surface treatment. The functionalized devices exhibit significant improvements in electronic output, stemming from the improved SLB quality, therefore strengthening the case for the use of such an approach in membrane‐on‐a‐chip systems.

Anti-Colonization Effect of Au Surfaces with Self-Assembled Molecular Monolayers Functionalized with Antimicrobial Peptides on S. epidermidis.

Medical devices with an effective anti-colonization surface are important tools for combatting healthcare-associated infections. Here, we investigated the anti-colonization efficacy of antimicrobial peptides covalently attached to a gold model surface. The gold surface was modified by a self-assembled polyethylene glycol monolayer with an acetylene terminus. The peptides were covalently connected to the surface through a copper-catalyzed [3 + 2] azide-acetylene coupling (CuAAC). The anti-colonization efficacy of the surfaces varied as a function of the antimicrobial activity of the peptides, and very effective surfaces could be prepared with a 6 log unit reduction in bacterial colonization.

Spatially Resolved Chemical Detection with a Nanoneedle-Probe-Supported Biological Nanopore.

In this article, we describe the quantitative characterization of a gold nanoneedle ion channel probe and demonstrate the utility of this probe for spatially resolved detection of a small molecule using ion channel activity. Our report builds on recent reports of Ide and co-workers, who reported the use of an etched gold wire modified with a poly(ethylene) glycol monolayer as a support for a lipid bilayer and subsequent single ion channel recordings. Although this nanoneedle electrode approach was reported previously, in our report, we investigate the effects of several operational parameters on the performance of the ion channel measurement and electrochemical phenomenon that occur in the nanoconfined space between the supported bilayer and the gold electrode. More specifically, we address the effects of length of the supporting monolayer and the composition of the electrolyte baths on channel current measurements and provide a quantitative description of what carries current at the working electrode (double-layer charging). In addition, we demonstrate the ability to control the direction of protein insertion (tip side vs bath side) with freely diffusing protein, which has not been previously reported, with the former method (tip side) enabling single-molecule detection of β-cyclodextrin (βCD) using a reconstituted α-hemolysin channel. Finally, anticipating future use of a nanoneedle-based biological nanopore probe in a scanned-probe microscopy, we demonstrate the ability to quantify and spatially resolve the concentration of βCD molecules in a microfluidic channel. We believe, in the long term, the described nanoneedle-based biological nanopore probe can be employed in, for example, scanning ion conductance microscopy using ion channels.

Functionalization of silicon surface through modification with polyethylene glycol monolayer

Functionalization of silicon surface through modification with polyethylene glycol monolayer

Assembly of citrate gold nanoparticles on hydrophilic monolayers

Self-assembled monolayers (SAMs) as model surfaces were linked onto planar gold films thorough lipoic acid or disulfide groups. The molecules used were polyethylene glycol (EG-S-S), N-[tris-(hydroxymethyl)methyl]acrylamide polymers with and without lipoic acid (Lipa-pTHMMAA and pTHMMAA) and a lipoic acid triazine derivative (Lipa-MF). All the layers, but Lipa-MF with a primary amino group were hydroxyl terminated. The layers were characterized by contact angle measurements and atomic force microscopy, AFM. Citrate stabilized nanoparticles, AuNPs in water and phosphate buffer were allowed to assemble on the layers for 10min and the binding was followed in real-time with surface plasmon resonance, SPR. The SPR resonance curves were observed to shift to higher angles and become increasingly damped, while also the peaks strongly broaden when large nanoparticles assembled on the surface. Both the angular shift and the damping of the curve was largest for nanoparticles assembling on the EG-S-S monolayer. High amounts of particles were also assembled on the pTHMMAA layer without the lipoic acid group, but the damping of the curve was considerably lower with a more even distribution of the particles. Topographical images confirmed that the highest number of particles were assembled on the polyethylene glycol monolayer. By increasing the interaction time more particles could be assembled on the surface.

Cytokines and growth factors cross-link heparan sulfate.

The glycosaminoglycan heparan sulfate (HS), present at the surface of most cells and ubiquitous in extracellular matrix, binds many soluble extracellular signalling molecules such as chemokines and growth factors, and regulates their transport and effector functions. It is, however, unknown whether upon binding HS these proteins can affect the long-range structure of HS. To test this idea, we interrogated a supramolecular model system, in which HS chains grafted to streptavidin-functionalized oligoethylene glycol monolayers or supported lipid bilayers mimic the HS-rich pericellular or extracellular matrix, with the biophysical techniques quartz crystal microbalance (QCM-D) and fluorescence recovery after photobleaching (FRAP). We were able to control and characterize the supramolecular presentation of HS chains—their local density, orientation, conformation and lateral mobility—and their interaction with proteins. The chemokine CXCL12α (or SDF-1α) rigidified the HS film, and this effect was due to protein-mediated cross-linking of HS chains. Complementary measurements with CXCL12α mutants and the CXCL12γ isoform provided insight into the molecular mechanism underlying cross-linking. Fibroblast growth factor 2 (FGF-2), which has three HS binding sites, was also found to cross-link HS, but FGF-9, which has just one binding site, did not. Based on these data, we propose that the ability to cross-link HS is a generic feature of many cytokines and growth factors, which depends on the architecture of their HS binding sites. The ability to change matrix organization and physico-chemical properties (e.g. permeability and rigidification) implies that the functions of cytokines and growth factors may not simply be confined to the activation of cognate cellular receptors.

UV/ozone surface treatment increases hydrophilicity and enhances functionality of SU-8 photoresist polymer

Abstract SU-8 photoresist polymer is widely used in the fabrication of microdevices. However, for biological applications, the problem of efficiently modifying SU-8 surfaces without perturbing roughness has not been successfully resolved. We present UV/ozone (UVO) surface pre-treatment as an effective method to increase the hydrophilicity of SU-8 films without affecting surface roughness, thus improving specific covalent binding of bio-molecules. We demonstrate that 30 s UVO treatment suffices to create carboxyl groups at the surface that can then be used for high density binding of molecules via amide bond formation. We further demonstrate that a two-step surface modification where the surface is first protected with an ethylene glycol monolayer leads to an increase in binding specificity. Finally, to illustrate the controlled binding and accessibility of immobilized molecules, we show three cycles of reversible interactions between anti-tamra antibody and tamra-cadaverine immobilized on the surface of SU-8.

Quantitative mechanical analysis of thin compressible polymer monolayers on oxide surfaces.

A clear understanding of the mechanical behavior of nanometer thick films on nanostructures, as well as developing versatile approaches to characterize their mechanical properties, are of great importance and may serve as the foundation for understanding and controlling molecular interactions at the interface of nanostructures. Here we report on the synthesis of thin, compressible polyethylene glycol (PEG) monolayers with a wet thickness of <20 nm on tin dioxide (SnO2) nanofibers through silane-based chemistries. Nanomechanical properties of such thin PEG films were extensively investigated using atomic force microscopy (AFM). In addition, tip-sample interactions were carefully studied, with different AFM tip modifications (i.e., hydrophilic and hydrophobic) and in different ionic solutions. We find that the steric forces dominate the tip-sample interactions when the polymer film is immersed in solution with salt concentrations similar to biological media (e.g., 1x phosphate buffer solution), while van der Waals and electrostatic forces have minimal contributions. A Dimitriadis thin film polymer compression model shows that the linear elastic regime is reproducible in the initial 50% indentation of these films which have tunable Young's moduli ranging from 5 MPa for the low molecular weight films to 700 kPa for the high molecular weight PEG films. Results are compared with the same PEG films deposited on silicon substrates which helped quantify the structural properties and understand the relationship between the structural and the mechanical properties of PEG films on the SnO2 fibers.

Photopatterning of self-assembled poly (ethylene) glycol monolayer for neuronal network fabrication

The ability to culture individual neurons and direct their connections on functional interfaces provides a platform for investigating information processing in neuronal networks. Numerous methods have been used to design ordered neuronal networks on microelectrode arrays (MEAs) for neuronal electrical activities recording. However, so far, no method has been implemented, which simultaneously provides high-resolution neuronal patterns and low-impedance microelectrode. To achieve this goal, we employed a chemical vapor-deposited, non-fouling poly (ethylene) glycol (PEG) self-assembled monolayer to provide a cell repellant background on the MEAs. Photolithography, together with plasma etching of the PEG monolayer, was used to fabricate different patterns on MEAs. No electrode performance degradation was observed after the whole process. Dissociated cortical neurons were cultured on the modified MEAs, and the patterns were maintained for more than 3 weeks. Spontaneous and evoked neuronal activities were recorded. All of the results demonstrate this surface engineering strategy allows successful patterning of neurons on MEAs, and is useful for future studies of information processing in defined neuronal networks on a chip.

Photoinduced Monolayer Patterning for the Creation of Complex Protein Patterns

This work investigates self-assembled monolayers that were formed from a glycol-terminated thiol monomer and were patterned using photoinduced monolayer desorption. Utilizing direct-write photolithography provided a facile means to generate complex protein patterns containing gradients and punctate regions. The ablated glycol monolayers were characterized using scanning probe microscopy, which allowed us to observe differences in the nanomechanical properties between the patterned and nonpatterned regions of the substrate. The patterned regions on the surface adsorbed proteins, and this process was monitored quantitatively using surface plasmon resonance imaging (SPRi). Moreover, the concentration of the protein could be controlled accurately by simply setting the gray level in the 8-bit image. Adsorbed protein was probed using a commercially available antibody binding assay, which showed significant enhancement over the background. The ability to produce complex protein patterns will contribute greatly to creating in vitro models that more accurately mimic an in vivo environment.

The label free picomolar detection of insulin in blood serum

Insulin, a polypeptide hormone secreted by pancreatic cells, is a key regulator in glucose homeostasis. Its deficiency leads to insulin-dependent (type I) diabetes whereas resistance to insulin is common in type II diabetes, obesity and a range of endocrine disorders. Its determination is of considerable value, particularly in the clinical diagnosis of diabetes mellitus and the doping control of athletes. It has, additionally, been noted as a potential breast cancer marker (serum insulin levels being found to be raised in comparison to control patients). Electrochemical assays are potentially very cheap, highly sensitive, and very readily transposed to a point of care. Though there exist numerous examples of label free impedimetric or capacitative assaying of biomolecules, these are rarely demonstrated to be effective in complex biological mixtures or to be applicable to low molecular weight targets (since they operate through the interfacial displacement of water/ions and/or the steric blocking of a redox probe). We report herein an ultrasensitive electrochemical and label-free biosensor for insulin in blood serum with a clinically relevant linear range and detection limit of 1.2pM. The transducing surfaces, based on readily prepared, antibody modified, polyethylene glycol monolayer modified polycrystalline gold surfaces, respond in a highly specific and re-useable manner to the target in up to 50% blood serum.

Direct patterning of coplanar polyethylene glycol alkylsilane monolayers by deep-ultraviolet photolithography as a general method for high fidelity, long-term cell patterning and culture.

This manuscript details a general method for patterning coplanar alkylsilane monolayers using deep-ultraviolet photolithography that has broad application for high fidelity patterning of cells of varying phenotype in long-term cultures. A polyethylene glycol monolayer was formed on a silica substrate and then patterned using 193 nm light from an ArF excimer laser. The regions of photoablation were then rederivatized with (3-trimethoxysilyl propyl) diethyltriamine (DETA), yielding high contrast cytophilic islands that promoted cell adhesion and growth. Rat hippocampal neurons, motoneurons, and myoblasts were then cultured in a defined, serum-free medium on the patterned surfaces for periods in excess of 40 days. This approach has been shown to be useful as a general method for the long-term culture of multiple cell types in highly defined spatial patterns and can be used for supporting complex cocultures for creating in vitro models for biological systems.

Nanoscale film structure related to capacitive effects in ethylene-glycol monolayers

A large improvement of the electrochemical capacitive detection of ssDNA and antigens by using ethylene-glycol alkanethiol monolayers has been recently demonstrated. Alkanethiol monolayers without ethylene-glycol functionalization do not show the required electrochemical stability. In this paper, we demonstrate that both phenomena are related to their morphological structure at nano-scale. Ethylene-glycol monolayers were compared with similar films without ethylene-glycol function. Capacitance measurements were used to verify the improved capacitance stability of the electrochemical interface. AFM images were used to investigate films packing and morphology at the nano-scale. This letter demonstrates that films without ethylene-glycol segment present deep grooves which are related to electrochemical instability while the improvement in case of ethylene-glycols is due to the absence of such a feature.

Cross-Linked Surface-Grafted Glycopolymer for Multivalent Recognition of Lectin

An alpha-link mannose-conjugated acrylamide monomer was synthesized. This monomer was polymerized by free radical polymerization with acrylamide, a cross-linker, and a surface linker directly on the gold surface. The surface linker, with an active carbon-carbon double bond, was preimmobilized on the gold surface by the thiol anchor. Thus, a cross-linked mannose-conjugated polymer thin layer was grafted onto a gold surface. This thin layer of polymer showed high binding sensitivity and excellent selectivity to its target lectin, concanavalin A (Con A), surpassing the formerly used linear glycopolymer and self-assembled glycol monolayers, validated by the techniques of quartz crystal microbalance, atomic force microscopy, and surface plasmon resonance. Remarkable response was observed to Con A at a concentration as low as 5 x 10(-10) M. The response is proportional to the Con A concentration up to 10(-7) M in phosphate-buffered saline. The use of cross-linked polymer decreased the flexibility of the polymer backbone between the carbohydrate binding sites. Therefore, the cost of conformational entropy for multivalent binding was minimized. The binding constants of the so-prepared cross-linked polymer with Con A were measured to be between 2.5 x 10(6) and 3.2 x 10(6) M(-1). These values are significantly larger than that obtained in our early study with a carbohydrate self-assembled monolayer. In addition to the carbohydrate-lectin recognition, additional selectivity may be achieved by controlling the degree of cross-linking.

Structure and DNA hybridization properties of mixed nucleic acid/maleimide-ethylene glycol monolayers.

The surface structure and DNA hybridization performance of thiolated single-strand DNA (HS-ssDNA) covalently attached to a maleimide-ethylene glycol disulfide (MEG) monolayer on gold have been investigated. Monolayer immobilization chemistry and surface coverage of reactive ssDNA probes were studied by X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry. Orientation of the ssDNA probes was determined by near-edge X-ray absorption fine structure (NEXAFS). Target DNA hybridization on the DNA-MEG probe surfaces was measured by surface plasmon resonance (SPR) to demonstrate the utility of these probe surfaces for detection of DNA targets from both purified target DNA samples and complex biological mixtures such as blood serum. Data from complementary techniques showed that immobilized ssDNA density is strongly dependent on the spotted bulk DNA concentration and buffer ionic strength. Variation of the immobilized ssDNA density had a profound influence on the DNA probe orientation at the surface and subsequent target hybridization efficiency. With increasing surface probe density, NEXAFS polarization dependence results (followed by monitoring the N 1s --> pi* transition) indicate that the immobilized ssDNA molecules reorient toward a more upright position on the MEG monolayer. SPR assays of DNA targets from buffer and serum showed that DNA hybridization efficiency increased with decreasing surface probe density. However, target detection in serum was better on the "high-density" probe surface than on the "high-efficiency" probe surface. The amounts of target detected for both ssDNA surfaces were several orders of magnitude poorer in serum than in purified DNA samples due to nonspecific serum protein adsorption onto the sensing surface.

Surface Phase Transition of C12E1 at the Air/Water Interface: A Study by Dynamic Surface Tension, External RA FT-IR, and 2D IR Correlation Methods

The surface conformational states of the Gibbs monolayer of ethylene glycol mono-n-dodecyl ether (C(12)E(1)) at the air/water interface was studied using dynamic surface tension, external reflection-absorption FT-IR spectroscopy (ERA FT-IR), and two-dimensional infrared (2D IR) correlation methods at constant temperature. The dynamic surface tensions were measured at different bulk concentrations of C(12)E(1), and it was observed that a constant surface tension region appears at approximately 38.5 mN m(-1) in a dynamic surface tension profile at concentrations higher than 11 micromol kg(-1). This constant surface tension region corresponds to the surface phase transition from liquid expanded (LE) to liquid condensed (LC). Two sets of ERA FT-IR spectra were collected, one at different bulk concentrations but after equilibrium time (equilibrium measurements) and another at constant bulk concentration (m = 16 micromol kg(-1)) but at different times (dynamic measurements). The first set of these measurements show that the peak area increases in the range of 11 < m < or = 16 micromol kg(-1), which means the increase in the number of surfactant molecules at the air/water interface. Also, the wavenumber of antisymmetric CH(2) stretching decreases gradually from approximately 2923 cm(-1) (for 10 and 11 micromol kg(-1)) to approximately 2918 cm(-1) (for m > or = 16 micromol kg(-1)) with increasing concentration. The wavenumbers of 2923 and 2918 cm(-1) were assigned to LE and LC phases, respectively, and the decrease of wavenumber in the concentration range of 11 < m < or = 16 micromol kg(-1) were correlated to the surface phase transition (LE --> LC), or in other words, in the mentioned concentration range, two phases coexist. The dynamic ERA FT-IR measurements at 16 micromol kg(-1) also confirm the surface phase transition from LE to LC. The 2D IR correlation method was applied to the both equilibrium and dynamic IR spectra of the C(12)E(1) monolayer. The synchronous correlation maps show two strong autopeaks at approximately 2922 and approximately 2851 cm(-1) and also show a strong correlation (cross-peaks) between antisymmetric CH(2) stretching (nu(a)) and symmetric CH(2) stretching (nu(s)). The asynchronous correlation maps show that both observed bands of nu(a) and nu(s) in one-dimensional IR split into two components with the characteristic of overlapped bands, which reveals the coexistence of two phases (LE and LC) at the interface at 11 < m < or = 16 micromol kg(-1). The synchronous and asynchronous maps that were obtained from dynamic IR spectra closely resembled the equilibrium map.

Penetration of Crystalline Powder Particles into Excised Human Skin Membranes and Model Gels from a Supersonic Powder Injector

The penetration of crystalline ibuprofen particles into excised human skin membranes and model target gels on actuation with the PowderJect ND1 ballistic needle-free injector has been examined. The deliverable dose of powder exiting the device is approximately 50% of the total cassette loading; the rest is lost via a gap forced open between the injector base and the surface of the target gel. It could be shown that substantial comminution of two different ibuprofen particle size fraction (38-53 microm, 53-75 microm) occurs within the injector on actuation. This resulted in an equiparation of the two initially different size fractions. Reduced comminution occurs with an ibuprofen/PVP (95:5) particulate formed by spray-drying/compaction/milling. On actuation into excised human skin membranes approximately 37% of cassette dose is recovered from the stratum corneum by stripping, and only 3% from the epidermis. It is concluded that powder delivery is mainly intra-epidermal with few particles fully breaching the stratum corneum. This is a consequence of the relative magnitude of the particle diameter (approximately 48 microm) and the stratum corneum thickness (approximately 10 microm). A 'soft' polyethylene glycol monolayer target gel gives the closest total percentage recovery of powder to that seen with the excised human skin membranes. Bilayer gels differentiated between surface impact/retention and penetration, but were difficult to handle. Powder penetration studies should therefore preferentially to be performed with human skin-membranes.

Ethylene glycol intercalation in MgAlCO3 hydrotalcite and its low-temperature intermediate phases

Three series of hydrotalcites (Mg:Al=2:1, 3:1, and 4:1) and their low-temperature intermediate phases were treated with ethylene glycol. The hydrotalcites with 3:1 and 4:1 Mg:Al ratios intercalated ethylene glycol, but the sample with the 2:1 ratio did not. However, a hydrotalcite intermediate phase with carbonate anions grafted into the hydroxide layers (HT-B), and its hydrated form (HT-B-r), did intercalate ethylene glycol, even in samples with 2:1 Mg:Al ratios, owing to the low charge density of this intermediate phase. In all treated samples, intercalation resulted in the formation of an ethylene glycol monolayer or bilayer between the hydroxide layers.The quantity of intercalated ethylene glycol in all samples increased as the Mg:Al ratio increased and when interlayer water was present. Low-temperature intermediate phases HT-B and HT-B-r incorporated with 20–40% more ethylene glycol than did samples with hydrotalcite structure.Moderate heat treatment (225 °C) of intercalates caused grafting of approximately one ethylene glycol molecule per Al cation. The grafting of ethylene glycol molecules and carbonate anions proceeded independently, resulting in the formation of samples with both organic and inorganic components.

Numerical Analysis of Nonionic Surfactant Monolayers at Water/Air Interfaces

The time evolution of a monododecyl pentaethylene glycol monolayer at the water−air interface is investigated using velocity rescaled NVT molecular dynamics. The model we consider consists of 40 surfactant molecules and 2350 water molecules enclosed in a periodic box. The time evolution of this system is ruled by the CFF91 force field that includes intra- and intermolecular degrees of freedom for both surfactant and water molecules. The interface we consider herein is hence described in all atomic detail. We discuss the initial condition problem and study the relaxation properties to stationarity. Transient regimes of self-assembly of the surfactant chains in entangled structures in both air and water are described. From the stationary configuration, we define the interface location, determine the mass distribution across the interface, and discuss the validity of the tilt angles notion when structural roughness is considered. Using pair distribution functions, we show that, besides typical tilt angles, monododecyl pentaethylene glycol molecules might also develop domains on the interface that suggest an in-plane orientational order.

Covalently Modified Silicon and Diamond Surfaces: Resistance to Nonspecific Protein Adsorption and Optimization for Biosensing

We report the direct covalent functionalization of silicon and diamond surfaces with short ethylene glycol (EG) oligomers via photochemical reaction of the hydrogen-terminated surfaces with terminal vinyl groups of the oligomers, and the use of these monolayers to control protein binding at surfaces. Photochemical modification of Si(111) and polycrystalline diamond surfaces produces EG monolayers linked via Si-C bond formation (silicon) or C-C bond formation (diamond). X-ray photoelectron spectroscopy was used to characterize the monolayer composition. Measurements using fluorescently labeled proteins show that the EG-functionalized surfaces effectively resist nonspecific adsorption of proteins. Additionally, we demonstrate the use of mixed monolayers on silicon and diamond and apply these surfaces to control specific versus nonspecific binding to optimize a model protein sensing assay.