Antonella Badia

Streptavidin Arrays as Supramolecular Architectures in Surface-Plasmon Optical Sensor Formats

Abstract We describe a series of interfacial biomolecular architectures employed for studies aimed at elucidating the structural and dynamical factors governing biorecognition and binding reactions between surface-attached biotin-derivatives and analogues and streptavidin from solution. We compare binding matrices based on mixed lipid monolayers transferred to a solid support from the water/air-interface and surface coatings based on binary mixed self-assembled monolayers. The analytical techniques used to monitor the various layers and the binding events are based on surface-plasmon spectroscopy and microscopy, in some cases in combination with electrochemical methods controlling, in particular, the surface potential of electrode surfaces in contact with aqueous electrolyte solutions. Finally, an example is given for the sensitivity enhancement obtainable by combining the field-enhancement of surface-plasmon resonances with fluorescence detection schemes.

Probing the different phases of self‐assembled monolayers on metal surfaces: Temperature dependence of the C–H stretching modes

Reflection‐absorption infrared spectroscopy at near grazing incidence is used to investigate the thermal stability of self‐assembled monolayers of alkyl thiols, HS–(CH2)n−1‐R, adsorbed on polycrystalline metal surfaces. The spectral features of the C–H stretching region have been carefully analyzed in the temperature range from 150 to 450 K. These features are highly sensitive to chain orientation and chain conformation. On gold, the chains are found to gradually untilt but remain largely all‐trans up to about 350 K. Above this temperature, a broad, irreversible transition to a liquidlike phase is observed, and which is characterized by a large number of gauche conformational defects. This overall behavior did not depend upon the chain length or terminal group for the systems studied. In contrast, significant differences were observed on a silver substrate.

Influence of hydrophobic alkylated gold nanoparticles on the phase behavior of monolayers of DPPC and clinical lung surfactant.

The effect of hydrophobic alkylated gold nanoparticles (Au NPs) on the phase behavior and structure of Langmuir monolayers of dipalmitoylphosphatidylcholine (DPPC) and Survanta, a naturally derived commercial pulmonary surfactant that contains DPPC as the main lipid component and hydrophobic surfactant proteins SP-B and SP-C, has been investigated in connection with the potential implication of inorganic NPs in pulmonary surfactant dysfunction. Hexadecanethiolate-capped Au NPs (C(16)SAu NPs) with an average core diameter of 2 nm have been incorporated into DPPC monolayers in concentrations ranging from 0.1 to 0.5 mol %. Concentrations of up to 0.2 mol % in DPPC and 16 wt % in Survanta do not affect the monolayer phase behavior at 20 °C, as evidenced by surface pressure-area (π-A) and ellipsometric isotherms. The monolayer structure at the air/water interface was imaged as a function of the surface pressure by Brewster angle microscopy (BAM). In the liquid-expanded/liquid-condensed phase coexistence region of DPPC, the presence of 0.2 mol % C(16)SAu NPs causes the formation of many small, circular, condensed lipid domains, in contrast to the characteristic larger multilobes formed by pure lipid. Condensed domains of similar size and shape to those of DPPC with 0.2 mol % C(16)SAu NPs are formed by compressing Survanta, and these are not affected by the C(16)SAu NPs. Atomic force microscopy images of Langmuir-Schaefer-deposited films support the BAM observations and reveal, moreover, that at high surface pressures (i.e., 35 and 45 mN m(-1)) the C(16)SAu NPs form honeycomb-like aggregates around the polygonal condensed DPPC domains. In the Survanta monolayers, the C(16)SAu NPs were found to accumulate together with the proteins in the liquid-expanded phase around the circular condensed lipid domains. In conclusion, the presence of hydrophobic C(16)SAu NPs in amounts that do not influence the π-A isotherm alters the nucleation, growth, and morphology of the condensed domains in monolayers of DPPC but not of those of Survanta. Systematic investigations of the effect of the interaction of chemically defined NPs with the lipid and protein components of lung surfactant on the physicochemical properties of surfactant films are pertinent to understanding how inhaled NPs impact pulmonary function.

Probing the Electrochemical Deposition and/or Desorption of Self-Assembled and Electropolymerizable Organic Thin Films by Surface Plasmon Spectroscopy and Atomic Force Microscopy

Abstract The combination of surface plasmon spectroscopy (SPS) and atomic force microscopy (AFM) with electrochemical methods (cyclic voltammetry, potential step) is used to (i) probe the film/substrate interaction in alkanethiol monolayers formed on gold surfaces and (ii) monitor the electrochemically driven deposition of organic molecules onto a metal surface. The reductive desorption of alkanethiols at single crystal and polycrystalline gold surfaces was investigated by SPS and AFM. These experiments demonstrate the possibility of desorbing a self-assembled monolayer at a well-defined potential with all the consequences for selective (re)-functionalization. The self-assembly of alkanethiols on gold under potential control was also monitored by SPS. The results show that the surface derivatization of gold electrodes can be actively controlled by the manipulation of the electrode potential. Finally, the immobilization of biotin on gold surfaces has been carried out by the electropolymerization of a water-soluble, biotinylated derivative of 3-hydroxyphenylacetic acid. The molecular recognition of the biotinylated polyphenol film by the bacterial protein streptavidin was monitored by SPS. The packing density of the biotin labels in the polymer film leads to a very fast diffusion-controlled docking of the streptavidin to the surface. These studies clearly prove the usefulness of electrochemically controlled deposition to produce ultrathin film organic surfaces with specific function.

Redox actuation of a microcantilever driven by a self-assembled ferrocenylundecanethiolate monolayer: an investigation of the origin of the micromechanical motion and surface stress.

The electrochemically induced motion of free-standing microcantilevers is attracting interest as micro/nanoactuators and robotic devices. The development and implementation of these cantilever-based actuating technologies requires a molecular-level understanding of the origin of the surface stress that causes the cantilever to bend. Here, we report a detailed study of the electroactuation dynamics of gold-coated microcantilevers modified with a model, redox-active ferrocenylundecanethiolate self-assembled monolayer (FcC(11)SAu SAM). The microcantilever transducer enabled the observation of the redox transformation of the surface-confined ferrocene. Oxidation of the FcC(11)SAu SAM in perchlorate electrolyte generated a compressive surface stress change of -0.20 +/- 0.04 N m(-1), and cantilever deflections ranging from approximately 0.8 microm to approximately 60 nm for spring constants between approximately 0.01 and approximately 0.8 N m(-1). A comparison of the charge-normalized surface stress of the FcC(11)SAu cantilever with values published for the electrochemical oxidation of polyaniline- and polypyrrole-coated cantilevers reveals a striking 10- to 100-fold greater stress for the monomolecular FcC(11)SAu system compared to the conducting polymer multilayers used for electroactuation. The larger stress change observed for the FcC(11)SAu microcantilever is attributable to steric constraints in the close-packed FcC(11)SAu SAM and an efficient coupling between the chemisorbed FcC(11)S- monolayer and the Au-coated microcantilever transducer (vs physisorbed conducting polymers). The microcantilever deflection vs quantity of electrogenerated ferrocenium obtained in cyclic voltammetry and potential step/hold experiments, as well as the surface stress changes obtained for mixed FcC(11)S-/C(11)SAu SAMs containing different populations of clustered vs isolated ferrocenes, have permitted us to establish the molecular basis of stress generation. Our results strongly suggest that the redox-induced deflection of a FcC(11)SAu microcantilever is caused by a monolayer volume expansion resulting from collective reorientational motions induced by the complexation of perchlorate ions to the surface-immobilized ferroceniums. The cantilever responds to the lateral pressure exerted by an ensemble of reorienting ferrocenium-bearing alkylthiolates upon each other rather than individual anion pairing events. This finding has general implications for using SAM-modified microcantilevers as (bio)sensors because it indicates that the cantilever responds to collective in-plane molecular interactions rather than reporting individual (bio)chemical events.

Atomic force microscopy studies of lateral phase separation in mixed monolayers of dipalmitoylphosphatidylcholine and dilauroylphosphatidylcholine

Atomic force microscopy imaging of dipalmitoylphosphatidylcholine (DPPC)/dilauroylphosphatidylcholine (DLPC) monolayers deposited onto alkanethiol modified-gold surfaces by the Langmuir–Schaefer technique was used to investigate domain formation in a binary system where phase separation arises from a difference in the alkyl chain lengths of the lipids. We have established how the condensed domain structure (shape and size) in DPPC/DLPC monolayers depends on the surface pressure and lipid composition. The mixed monolayers exhibit a positive deviation from an ideal mixing behavior at surface pressures of ⩽32 mN/m. Lateral compression to pressures greater than the liquid-expanded-to-liquid-condensed (LE-to-LC) phase transition pressure of the mixed monolayer (∼8–16 mN/m) induces extensive separation into condensed DPPC-rich domains and a fluid DLPC matrix. The condensed structures observed at a few milliNeutons per meter above the LE-to-LC transition pressure resemble those reported for pure DPPC monolayers in the LE/LC co-existence region. At a bilayer equivalence pressure of 32 mN/m and 20 °C, condensed domains exist between xDPPC ∼0.25 and ∼0.80, analogous to aqueous DPPC/DLPC dispersions. Compression from 32 to 40 mN/m results in either a striking distortion of the DPPC domain shape or a break-up of the microscopic DPPC domains into a network of nanoscopic islands (at higher DPPC mol fractions), possibly reflecting a critical mixing behavior. The results of this study provide a fundamental framework for understanding and controlling the formation of lateral domain structures in mixed phospholipid monolayers.

Monolayer/Bilayer Transition in Langmuir Films of Derivatized Gold Nanoparticles at the Gas/Water Interface: An X-Ray Scattering Study

The microscopic structure of Langmuir films of derivatized gold nanoparticles has been studied as a function of area/particle on the water surface. The molecules (AuSHDA) consist of gold particles of mean core diameter D approximately 22 angstroms that have been stabilized by attachment of carboxylic acid terminated alkylthiols, HS-(CH2)15-COOH. Compression of the film results in a broad plateau of finite pressure in the surface pressure versus area/particle isotherm that is consistent with a first-order monolayer/bilayer transition. X-ray specular reflectivity (XR) and grazing incidence diffraction show that when first spread at large area/particle, AuSHDA particles aggregate two dimensionally to form hexagonally packed monolayer domains at a nearest-neighbor distance of a = 34 angstroms. The lateral positional correlations associated with the two-dimensional (2D) hexagonal order are of short range and extend over only a few interparticle distances; this appears to be a result of the polydispersity in particle size. Subsequent compression of the film increases the surface coverage by the monolayer but has little effect on the interparticle distance in the close-packed domains. The XR and off-specular diffuse scattering (XOSDS) results near the onset of the monolayer/bilayer coexistence plateau are consistent with complete surface coverage by a laterally homogeneous monolayer of AuSHDA particles. On the high-density side of the plateau, the electron-density profile extracted from XR clearly shows the formation of a bilayer in which the newly formed second layer on top is slightly less dense than the first layer. In contrast to the case of the homogeneous monolayer, the XOSDS intensities observed from the bilayer are higher than the prediction based on the capillary wave model and the assumption of homogeneity, indicating the presence of lateral density inhomogeneities in the bilayer. According to the results of Bragg rod measurements, the 2D hexagonal order in the two layers of the bilayer are only partially correlated.

The Effect of Terminal Hydrogen Bonding on the Structure and Dynamics of Nanoparticle Self-Assembled Monolayers (SAMs): An NMR Dynamics Study.

conditions. The sample shown in Figure 1a was prepared by heating the solution for 3 h at 80 C and casting the hot solution on a carbon-covered microscopy grid. Gray disks 5 nm in diameter were seen, which were identified as TiO2 particles. The particles agglomerated into chains consisting of approximately 20 individual disks. Apparently the heating procedure had two effects involved in the film formation. First TiO2 particles were formed in each block copolymer micelle. In a second step, the increased temperature caused the excess HCl to evaporate. The excess HCl is known, however, to be essential for stabilizing inverse diblock copolymer micelles during film formation. [10,11] As the HCl evaporates, the PEO block loses its ionic character and the kinetic as well as the thermodynamic stability of the diblock copolymer micelles is reduced because of the increased compatibility of the constituent blocks. Casting of films by simply allowing a drop of the solution to evaporate on a carbon-coated copper grid enabled the diblock copolymer micelles to react to the changing concentration and to transform from globular to worm-like micelles.

Nanostrand Formation of Block Copolymers at the Air/Water Interface

Langmuir-Blodgett monolayers consisting of a network of nanostrands have occasionally been reported in the literature, but are often coexistent with other morphologies, which is not useful for potential applications. With the use of PS-P4VP/PDP, a polystyrene-poly(4-vinyl pyridine) diblock copolymer of 12 mol % VP content mixed with 3-pentadecylphenol, it is shown that the disordered nanostrand network morphology can be obtained reproducibly and uniformly over large surface areas by spreading chloroform solutions of relatively high copolymer concentration. Use of a more slowly evaporating spreading solvent, 1,1,2,2-tetrachloroethane, and a low subphase temperature, 8-9 °C, results in much more densely aligned nanostrands. Poorly spreading solvents such as nitrobenzene produce the well-known fingerprint pattern often observed in spin- or dip-coated thin films of block copolymers. A mechanism for nanostrand network formation is proposed that involves the momentary formation of a fingerprint morphology in spreading drops followed by its breakup at the level of the mobile P4VP/PDP stripes as spreading continues, leaving P4VP-anchored PS nanostrands floating on the water surface.

Gold(I)-dithioether supramolecular polymers: synthesis, characterization, and luminescence.

A series of discrete compounds and supramolecular polymers were synthesized by self-assembly of dithioether building blocks and HAuCl4.3H2O. In complexes 1 {[AuL(1-Me)Cl], where L(1-Me) is bis(methylthio)methane} and 2 {[Au2L(2-Ph)Cl2], where L(2-Ph) is 1,2-bis(phenylthio)ethane}, adjacent units are connected via aurophilic interactions. Complex 1, a one-dimensional (1D) supramolecular polymer, and complex 2, a two-dimensional supramolecular network, both feature nearly linear [Au-Au-](infinity) chains. Complexes 4a, 4b, and 4c, all of which contain 1,3-bis(phenylthio)propane (L(3-Ph)), are polymorphs having the composition [Au2L(3-Ph)Cl2]. Complex 3 {[Au2L(1-Ph)Cl2], where L(1-Ph) is bis(phenylthio)methane}and complexes 4a and 4b consist of nearly identical 1D supramolecular polymers formed through Au-Au interactions. The third polymorph, 4c, is a molecular complex, as it does not have metal-metal interactions. Complex 5 {[Au2L(4-Ph)Cl2], where L(4-Ph) is 1,4-bis(phenylthio)butane} is also molecular. UV-vis spectra showed that the absorption bands of these complexes are allowed ligand-centered transitions between 230 and 260 nm. Complexes 1, 2, and 6 {[AuL(3-Me)Cl], where L(3-Me) is 1,3-bis(methylthio)propane} exhibited solid-state luminescence at 5 K with vibronic progressions and band maxima at approximately 570 nm. It is suggested that complex 6 contains [Au-Au-](infinity) chains.