Gunnar Dunér

Optimizing immobilization on two-dimensional carboxyl surface: pH dependence of antibody orientation and antigen binding capacity

The performance of immunosensors is highly dependent on the amount of immobilized antibodies and their remaining antigen binding capacity. In this work, a method for immobilization of antibodies on a two-dimensional carboxyl surface has been optimized using quartz crystal microbalance biosensors. We show that successful immobilization is highly dependent on surface pK(a), antibody pI, and pH of immobilization buffer. By the use of EDC/sulfo-NHS (1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysulfosuccinimide) activation reagents, the effect of the intrinsic surface pK(a) is avoided and immobilization at very low pH is therefore possible, and this is important for immobilization of acidic proteins. Antigen binding capacity as a function of immobilization pH was studied. In most cases, the antigen binding capacity followed the immobilization response. However, the antigen-to-antibody binding ratio differed between the antibodies investigated, and for one of the antibodies the antigen binding capacity was significantly lower than expected from immobilization in a certain pH range. Tests with anti-Fc and anti-Fab(2) antibodies on different antibody surfaces indicated that the orientation of the antibodies on the surface had a profound effect on the antigen binding capacity of the immobilized antibodies.

Quartz Crystal Microbalance with Dissipation (QCM-D) studies of the viscoelastic response from a continuously growing grafted polyelectrolyte layer

Poly(acrylic acid) was grown from substrates by photopolymerization, and the grafting process was monitored in situ by Quartz Crystal Microbalance with Dissipation (QCM-D) measurements in a 1:1 v/v mixture of water/ethanol. The polymerization process was monitored into the thick film region, where the change in frequency and dissipation with increasing film mass changes sign as predicted by the Voigt viscoelastic model. Our experimental data are compared with predictions of this model, and satisfactory agreement is found for low overtone numbers. The Voigt model was applied to analyze the measured changes in frequency, Δf, and dissipation, ΔD, in order to extract information on layer thickness, shear elasticity, μ, and shear viscosity, η, of the growing film. The increasing rate of changes in Δf and ΔD observed after about 150s of polymerization was found to correlate with an increasing growth rate of the film thickness. For longer polymerization times a close to linear increase in thickness with time was observed. The sensitivity, defined as the derivatives of Δf and ΔD with respect to thickness, depends on overtone number and is different for the frequency and dissipation signals - facts that should be considered when investigating small changes in thick films used in e.g. sensor applications.

Nanomechanical mapping of a high curvature polymer brush grafted from a rigid nanoparticle

Analysis of interaction forces when probing a silica core–polyelectrolyte brush shell nanoparticle, adsorbed on a silica substrate and bathed by aqueous electrolyte solution, with an ultrasharp atomic force microscopy (AFM) tip provides a spatially resolved map of heterogeneous mechanical properties across the nanoparticle. The deformation of the brush is mainly compressive when probed directly above the nanoparticle centre and mainly deflective when probed at a finite horizontal distance away from the centre. The brush is significantly stiffer against compression than against deflection, and ionization of the brush has a greater stiffening effect against compression than deflection. Whereas a height image of the core–shell nanoparticle was unremarkable, showing a monotonic decrease in height with increasing horizontal distance from the centre, brush deformation, energy dissipation and adhesion displayed local minima over the centre and maxima at a finite horizontal distance away from the centre, corresponding to a position near the rigid core nanoparticle edge. The different response to brush deformation depending on the angle of probing is relevant to the interactions of brush-decorated macroscopic surfaces with submicrometer roughness and to the interactions of brush-decorated nanoparticles with ultrafine structures in their environments.

Surface-confined photopolymerization of pH-responsive acrylamide/acrylate brushes on polymer thin films

Dynamic acrylamide/acrylate polymeric brushes were synthesized at gold-plated quartz crystal surfaces. The crystals were initially coated with polystyrene-type thin films, derivatized with photolabile iniferter groups, and subsequently subjected to photoinitiated polymerization in acrylamide/acrylate monomer feeds. This surface-confined polymerization method enabled direct photocontrol over the polymerization, as followed by increased frequency responses of the crystal oscillations in a quartz crystal microbalance (QCM). The produced polymer layers were also found to be highly sensitive to external acid/base stimuli. Large oscillation frequency shifts were detected when the brushes were exposed to buffer solutions of different pH. The dynamic behavior of the resulting polymeric brushes was evaluated, and the extent of expansion and contraction of the films was monitored by the QCM setup in situ in real time. The resulting responses were rapid, and the effects were fully reversible. Low pH resulted in full contractions of the films, whereas higher pH yielded maximal expansion in order to minimize repulsion around the charged acrylate centers. The surfaces also proved to be very robust because the responsiveness was reproducible over many cycles of repeated expansion and contraction. Using ellipsometry, copolymer layers were estimated to be approximately 220 nm in a collapsed state and approximately 340 nm in the expanded state, effectively increasing the thickness of the film by 55%.

Letter to the Editor: Friction between Surfaces—Polyacrylic Acid Brush and Silica—Mediated by Calcium Ions

With this letter, we report how friction can be controlled by inducing physical bonds solely within a polyelectrolyte brush layer, while keeping repulsive interactions between the brush layer and the bare surface that slides above. Our results imply that the nature of the bare surface is of minor importance as long as the repulsive surface interaction is maintained.

Reducing Protein Adsorption with Polymer-Grafted Hyaluronic Acid Coatings

We report a thermoresponsive chemical modification strategy of hyaluronic acid (HA) for coating onto a broad range of biomaterials without relying on chemical functionalization of the surface. Poly(di(ethylene glycol) methyl ether methacrylate) (PMEO2MA), a polymer with a lower critical solution temperature of 26 °C in water, was grafted onto HA to allow facile formation of biopolymer coatings. While the mechanism for film formation appears to involve a complex combination of homogeneous nucleation followed by heterogeneous film growth, we demonstrate that it resulted in hydrophilic coatings that significantly reduce protein adsorption despite the high fraction of hydrophobic (PMEO2MA). Structural characterization was performed using atomic force microscopy (AFM), which showed the formation of a dense, continuous coating based on 200 nm domains that were stable in protein solutions for at least 15 days. The coatings had a water contact angle of 16°, suggesting the formation of hydrophilic but not fully wetting films. Quartz crystal microbalance with dissipation monitoring (QCM-D) as well as biolayer interferometry (BLI) techniques were used to measure adsorption of bovine serum albumin (BSA), fibrinogen (Fbg), and human immunoglobulin (IgG), with results indicating that HA-PMEO2MA-coated surfaces effectively inhibited adsorption of all three serum proteins. These results are consistent with previous studies demonstrating that this degree of hydrophilicity is sufficient to generate an effectively nonfouling surface and suggest that segregation during the solubility transition resulted in a surface that presented the hydrophilic HA component of the hybrid biopolymer. We conclude that PMEO2MA-grafted HA is a versatile platform for the passivation of hydrophobic biomaterial surfaces without need for substrate functionalization.

Attractive double-layer forces and charge regulation upon interaction between electrografted amine layers and silica

Amine functionalities have been introduced on glassy carbon surfaces through electrografting of 4-(2-aminoethyl)benzenediazonium tetrafluoroborate. The grafted layers were characterized by ellipsometry and by nanomechanical mapping in air and aqueous solutions using the atomic force microscopy PeakForce QNM mode. The layer was found to be 2.5 nm thick with low roughness, comparable to that of the glassy carbon substrate. However, small semi-spherical features were observed in the topographical image, indicating a clustering of the grafted amine compound. The nanomechanical mapping also demonstrated some swelling of the layer in water and pointed toward an important contribution of electrostatic interactions for the tip-surface adhesion. The forces between an aminated glassy carbon surface and a μm-sized silica particle in aqueous solutions were measured at different ionic strength and pH-values. The results demonstrate that an attractive double-layer force predominates at large separations, and that the surface charge densities increase as the separation between the surfaces decreases. The degree of charge regulation on the aminated glassy carbon is significant. The relatively low surface charge density of the aminated glassy carbon is attributed to significant incorporation of counterions in the water-rich grafted layer.

Transient Marangoni transport of colloidal particles at the liquid/liquid interface caused by surfactant convective-diffusion under radial flow

HYPOTHESIS Interfacial tension gradients at a liquid/liquid interface drive Marangoni flows. When colloidal particles are adsorbed to an interface in systems with spatial and temporal gradients of surfactant concentration, these interfacial flows can be potentially significant contributors to the direction and rate of particle transport. EXPERIMENTS In this work, we use optical microscopy to measure the interfacial velocities of 5μm diameter polystyrene latex particles adsorbed at an oil/water interface, using olive oil to represent polar oils often encountered in cleaning applications. FINDINGS On surfactant adsorption the maximum interfacial velocity scales linearly with bulk surfactant concentration, even for concentrations exceeding the critical micelle concentration (CMC). The maximum interfacial velocity weakly decreases with increasing flow rate, but it varies non-monotonically with the radial distance from the inlet. Upon surfactant desorption into a rinse solution, the maximum velocity increases with increasing concentration of the original surfactant solution, but only up to a plateau near the CMC. These experimental trends are well-described by a convective-diffusion model for surfactant transport to or from the liquid/liquid interface coupled with Langmuir-type adsorption, using a constitutive relation between the interfacial tension gradient and interfacial velocity based on the interfacial tangential stress jump.

Effect of polyelectrolyte–surfactant complexation on Marangoni transport at a liquid–liquid interface

Complexation of surfactants and oppositely charged polyelectrolytes is expected to alter Marangoni transport at a fluid interface compared to either single component system due to altered interfacial tension isotherms and mass transfer rates as well as adsorption irreversibility effects. We investigate Marangoni transport at the oil/water interface by passing mixtures of the anionic surfactant sodium dodecyl sulfate (SDS) and cationic polyelectrolyte poly(3-(2-methylpropionamide)propyl) trimethylammonium chloride-acrylamide (poly[AM-MAPTAC]), or rinsing solutions, over an oil/water interface in a radial, stagnation point flow. The displacements of adsorbed tracer particles are recorded through optical microscopy. The net displacement, defined as the sum of the displacements occurring during the adsorption and desorption stages of one application and rinsing cycle, is up to 10 times greater for complexing surfactant/polymer mixtures compared to either single component system. The enhanced net displacement is largely determined by the enhanced transport upon adsorption, while the reverse displacement that would normally occur upon rinsing is partially suppressed by partially irreversible polymer adsorption at the oil/water interface. In addition to effects of complexation on interfacial tension gradient induced flow, complexation effects on the bulk, and possibly interfacial, viscosity also influence the interfacial transport.