Zwitterionic silica nanogel-modified polysulfone nanoporous membranes formed by in-situ method for water treatment.

We report a simple methodology to prepare nano-porous polysulfone membranes using zwitterionic functionalized silica nanogels with high BSA protein rejection and antifouling properties. The zwitterionic silica precursor was prepared by reacting 1,3-propane sultone with 3-aminopropyl triethoxysilane under an inert atmosphere. The precursor was in situ hydrolyzed and condensed in the polysulfone nanoporous membrane network by one-pot acidic phase inversion. The prepared membranes were characterized to establish their physicochemical nature, morphology, and basic membrane properties such as permeation, rejection, and recovery. The zwitterionic membranes showed improved hydrophilicity, membrane water uptake (∼83.5%), water permeation, BSA protein rejection (>95%), and dye rejection (congo red: >52% (∼6-fold increase); methylene blue: ∼15% (∼2-fold increase)) were improved without compromising the membrane flux and fouling resistance. Overall, we report an easy fabrication method of efficient nanocomposite zwitterionic ultrafilter membranes for water treatment with excellent flux, protein separation, filtration efficiency, and antifouling behavior.

Catechol Containing Polyelectrolyte Complex Nanoparticles as Local Drug Delivery System for Bortezomib at Bone Substitute Materials.

The proteasome inhibitor bortezomib (BZM) is one of the most potent anti-cancer drugs in the therapy of multiple myeloma. In this study, an adhesive drug delivery system (DDS) for BZM was developed. Therefore, we extended the present DDS concept of polyelectrolyte complex (PEC) nanoparticle (NP) based on electrostatic interactions between charged drug and polyelectrolyte (PEL) to a DDS concept involving covalent bonding between PEL and uncharged drugs. For this purpose, 3,4-dihydroxyphenyl acetic acid (DOPAC) was polymerized via an oxidatively induced coupling reaction. This novel chemo-reactive polyanion PDOPAC is able to temporarily bind boronic acid groups of BZM via its catechol groups, through esterification. PDOPAC was admixed to poly(l-glutamic acid) (PLG) and poly(l-lysine) (PLL) forming a redispersible PEC NP system after centrifugation, which is advantageous for further colloid and BZM loading processing. It was found that the loading capacity (LC) strongly depends on the PDOPAC and catechol content in the PEC NP. Furthermore, the type of loading and the net charge of the PEC NP affect LC and the residual content (RC) after release. Release experiments of PDOPAC/PEC coatings were performed at medically relevant bone substitute materials (calcium phosphate cement and titanium niobium alloy) whereby the DDS worked independently of the surface properties. Additionally, in contrast to electrostatically based drug loading the release behavior of covalently bound, uncharged BZM is independent of the ionic strength (salt content) in the release medium.

Nanoparticle assembly under block copolymer confinement: The effect of nanoparticle size and confinement strength.

We investigate the self-assembly of cylinder-forming polystyrene-block-poly(4-vinylpyridine) (PS-b-P4VP) block copolymers (BCP) mixed with metal nanoparticles (NP) coated with short-chain polystyrene (PS) ligands. The NP formed hierarchical superstructures under confinement of cylindrical PS domains of PS-b-P4VP BCP. The complexity of NP superstructures was found to depend on the ratio between PS cylindrical domain size and NP size (DC/DNP). As the DC/DNP ratio increased, the number of NP layers normal to the cylinder axis also increased. However, the packing density of the NP decreased at higher DC/DNP. Furthermore, the morphology of the structures obtained during different solvent casting conditions revealed that the initial clustering of NP and micellization around these clusters act as a precursor for the subsequent formation of closely packed structures of NP in cylinders. The experimental results were further supported by modeling results obtained from molecular dynamics (MD) simulation. Based on MD simulations, we constructed structural phase diagram of nanoparticle assemblies in the presence of asymmetric diblock copolymers comprising short NP-attractive blocks. The MD simulation results indicate that NP undergo transition from spherical to cylindrical assemblies depending on the NP size, the overall concentration of components and the degree of affinity of the minor block to NP.

In-situ-Investigation of Enzyme Immobilization on Polymer Brushes.

Herein, we report on the use of a combined setup of quartz-crystal microbalance, with dissipation monitoring and spectroscopic ellipsometry, to comprehensively investigate the covalent immobilization of an enzyme to a polymer layer. All steps of the covalent reaction of the model enzyme glucose oxidase with the poly(acrylic acid) brush by carbodiimide chemistry, were monitored in-situ. Data were analyzed using optical and viscoelastic modeling. A nearly complete collapse of the polymer chains was found upon activation of the carboxylic acid groups with N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide and N-Hydroxysuccinimide. The reaction with the amine groups of the enzyme occurs simultaneously with re-hydration of the polymer layer. Significantly more enzyme was immobilized on the surface compared to physical adsorption at similar conditions, at the same pH. It was found that the pH responsive swelling behavior was almost not affected by the presence of the enzyme.

Free Polyethylenimine Enhances Substrate-Mediated Gene Delivery on Titanium Substrates Modified With RGD-Functionalized Poly(acrylic acid) Brushes.

Substrate mediated gene delivery (SMD) is a method of immobilizing DNA complexes to a substrate via covalent attachment or nonspecific adsorption, which allows for increased transgene expression with less DNA compared to traditional bolus delivery. It may also increase cells receptivity to transfection via cell-material interactions. Substrate modifications with poly(acrylic) acid (PAA) brushes may improve SMD by enhancing substrate interactions with DNA complexes via tailored surface chemistry and increasing cellular adhesion via moieties covalently bound to the brushes. Previously, we described a simple method to graft PAA brushes to Ti and further demonstrated conjugation of cell adhesion peptides (i.e., RGD) to the PAA brushes to improve biocompatibility. The objective of this work was to investigate the ability of Ti substrates modified with PAA-RGD brushes (PAA-RGD) to immobilize complexes composed of branched polyethyleneimine and DNA plasmids (bPEI-DNA) and support SMD in NIH/3T3 fibroblasts. Transfection in NIH/3T3 cells cultured on bPEI-DNA complexes immobilized onto PAA-RGD substrates was measured and compared to transfection in cells cultured on control surfaces with immobilized complexes including Flat Ti, PAA brushes modified with a control peptide (RGE), and unmodified PAA. Transfection was two-fold higher in cells cultured on PAA-RGD compared to those cultured on all control substrates. While DNA immobilization measured with radiolabeled DNA indicated that all substrates (PAA-RGD, unmodified PAA, Flat Ti) contained nearly equivalent amounts of loaded DNA, ellipsometric measurements showed that more total mass (i.e., DNA and bPEI, both complexed and free) was immobilized to PAA and PAA-RGD compared to Flat Ti. The increase in adsorbed mass may be attributed to free bPEI, which has been shown to improve transfection. Further transfection investigations showed that removing free bPEI from the immobilized complexes decreased SMD transfection and negated any differences in transfection success between cells cultured on PAA-RGD and on control substrates, suggesting that free bPEI may be beneficial for SMD in cells cultured on bPEI-DNA complexes immobilized on PAA-RGD grafted to Ti. This work demonstrates that substrate modification with PAA-RGD is a feasible method to enhance SMD outcomes on Ti and may be used for future applications such as tissue engineering, gene therapy, and diagnostics.

Enzyme Immobilization in Polyelectrolyte Brushes: High Loading and Enhanced Activity Compared to Monolayers.

Catalysis by enzymes on surfaces has many applications. However, strategies for efficient enzyme immobilization with preserved activity are still in need of further development. In this work, we investigate polyelectrolyte brushes prepared by both grafting-to and grafting-from with the aim to achieve high catalytic activity. For comparison, self-assembled monolayers that bind enzymes with the same chemical interactions are included. We use the model enzyme glucose oxidase and two kinds of polymers: anionic poly(acrylic acid) and cationic poly(diethylamino)methyl methacrylate. Surface plasmon resonance and spectroscopic ellipsometry are used for accurate quantification of surface coverage. Besides binding more enzymes, the "3D-like" brush environment enhances the specific activity compared to immobilization on self-assembled monolayers. For grafting-from brushes, multilayers of enzymes were spontaneously and irreversibly immobilized without conjugation chemistry. When the pH was between the pI of the enzyme and the p Ka of the polymer, binding was considerable (thousands of ng/cm2 or up to 50% of the polymer mass), even at physiological ionic strength. However, binding was observed also when the brushes were neutrally charged. For acidic brushes (both grafting-to and grafting-from), the activity was higher for covalent immobilization compared to noncovalent. For grafting-from brushes, a fully preserved specific activity compared to enzymes in the liquid bulk was achieved, both with covalent (acidic brush) and noncovalent (basic brush) immobilization. Catalytic activity of hundreds of pmol cm-2 s-1 was easily obtained for polybasic brushes only tens of nanometers in dry thickness. This study provides new insights for designing functional interfaces based on enzymatic catalysis.

Fluorocarbon-Free Dual-Action Textile Finishes Based on Covalently Attached Thermoresponsive Block Copolymer Brush Coatings.

We present fluorocarbon-free block copolymer brushes as potential systems for dual-action, i.e., soil-repellent and soil-releasing textile finishes. Polymer brushes were prepared by employing specifically engineered triblock copolymers consisting of a hydrophobic, a hydrophilic, and either a central or a terminal anchor block bearing several anchoring groups for sustainable immobilization using the "grafting to" approach on both flat Si wafers and rough cotton fabrics. The switching characteristics of both types of block copolymer brushes were investigated by exposing the brushes to conditions and stimuli that are similar to those applied during laundering in a washing machine and drying in a laundry dryer, respectively. Contact angle measurements were performed to evaluate the polarity and wettability of the block copolymer brushes after treatment in hot water and in air, or in a vacuum at elevated temperatures simulating the washing and the drying procedure of a textile fabric. While the block copolymer brush with the terminal anchor showed only minor changes in terms of the wetting characteristics and the brush morphology upon the applied stimuli, the block copolymer brush with the central anchoring block exhibited a significant change from a hydrophilic (soil-releasing) to a hydrophobic (stain-repellent) surface. This switching behavior was reversible and could be achieved on both, flat Si wafers, and much more pronounced on rough cotton fabrics. Atomic force microscopy and angle-resolved X-ray photoelectron spectroscopy investigations further indicated a complete rearrangement of the brush morphology. Accordingly, we regard this type of block copolymer brushes as a system that fully meets the basic requirements for an application as a dual-action textile finish, which can be reversibly switched with respect to water repulsion.

Enzyme immobilization on protein-resistant PNIPAAm brushes: impact of biotin linker length on enzyme amount and catalytic activity.

Thermoresponsive poly(N-isopropylacrylamide) (PNIPAAm) brushes with terminal click functionality can be used to selectively immobilize enzymes. Exploiting their inherent protein-repellent and thus non-fouling properties, surfaces with specific bioactivity can be created in this way. This report describes the functionalization of alkyne-PNIPAAm brushes with two biotin linkers of different poly(ethylene glycol) (PEG) spacer length via click chemistry and the subsequent immobilization of streptavidin-conjugated horseradish peroxidase (SA-HRP) by the strong interaction between biotin and streptavidin. Spectroscopic ellipsometry is used to quantify the biotin and the SA-HRP amount on the PNIPAAm brushes. Enzyme activities are determined by UV­vis spectroscopy. A better accessibility to the alkyne-functionalized chain ends for the short biotin-PEG3 linker leads to a higher biotin amount on these PNIPAAm brushes, which in turn results in higher SA­HRP amounts on biotin-PEG3-modified brushes in the swollen (20 °C) and collapsed state (37 °C) compared to biotin-PEG23-modified ones. For both linkers, an increased immobilization temperature leads to higher SA-HRP amounts due to an enhanced reaction kinetics and mobility. All immobilized SA-HRP amounts are in range of 81-98% monolayer coverage, except for SA-HRP immobilized on biotin-PEG23-modified PNIPAAm brushes at 20 °C, exhibiting only 43% monolayer coverage. A high mobility of the biotin linker combined with a low surface loading of SA-HRP is found to be beneficial for the activity of SA-HRP. Hence, the highest specific activity as measured by HRP-catalyzed 3,3´,5,5´­tetramethylbenzidine (TMB) oxidation is found for a low SA-HRP amount on biotin-PEG23-modified brushes.

Biofunctionalization of Titanium Substrates Using Nanoscale Polymer Brushes with Cell Adhesion Peptides.

The grafting of polymer brushes to substrates is a promising method to modify surface properties such as wettability and the affinity toward proteins and cells for applications in microelectronics, biomedical devices, and sensors. Poly(acrylic) acid (PAA) brushes are of high interest because of their stimuli-responsive behavior and the presence of carboxy (COOH) groups, which allow for immobilization of bioactive molecules. The "grafting-to" approach results in homogeneous and well-defined polymer brushes, but, although grafting-to has been demonstrated with PAA brushes on silicon (Si) substrates, it has not been performed on biocompatible materials such as titanium (Ti). Here, we have described a facile method to modify biocompatible Ti substrates with PAA brushes to amplify their substrate functionality. The grafting-to PAA "pseudo" brushes were successfully grafted to Ti substrates and retained their pH-dependent swelling behavior. An RGD peptide was covalently bound to COOH groups of the PAA brushes (PAA-RGD) as a model bioactive group. While NIH/3T3 cell adhesion was significantly decreased on PAA-functionalized Ti substrates, PAA-RGD on Ti had cell adhesion comparable to that of flat Ti at 24 and 48 h, with significantly more cells adhered to PAA-RGD compared to PAA on Ti at 48 h.

Bioinspired thermoresponsive nanoscaled coatings: Tailor-made polymer brushes with bioconjugated arginine-glycine-aspartic acid-peptides.

The development of bioengineered surface coatings with stimuli-responsive properties is beneficial for a number of biomedical applications. Environmentally responsive and switchable polymer brush systems have a great potential to create such smart biointerfaces. This study focuses on the bioconjugation of cell-instructive peptides, containing the arginine-glycine-aspartic acid tripeptide sequence (RGD motif), onto well-defined polymer brush films. Herein, the highly tailored end-grafted homo polymer brushes are either composed of the polyelectrolyte poly(acrylic) acid (PAA), providing the reactive carboxyl functionalities, or of the temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm). Of particular interest is the preparation of grafted-to binary brushes using both polymers and their subsequent conversion to RGD-biofunctionalized PNIPAAm-PAA binary brushes by a carbodiimide conjugation method. The bioconjugation process of two linear RGD-peptides Gly-Arg-Gly-Asp-Ser and Gly-Arg-Gly-Asp-Ser-Pro-Lys and one cyclic RGD-peptide cyclo(Arg-Gly-Asp-D-Tyr-Lys) is comparatively investigated by complementary analysis methods. Both techniques, in situ attenuated total reflectance Fourier transform infrared spectroscopy measurements and the in situ spectroscopic ellipsometric analysis, describe changes of the brush surface properties due to biofunctionalization. Besides, the bound RGD-peptide amount is quantitatively evaluated by ellipsometry in comparison to high performance liquid chromatography analysis data. Additionally, molecular dynamic simulations of the RGD-peptides themselves allow a better understanding of the bioconjugation process depending on the peptide properties. The significant influence on the bioconjugation result can be derived, on the one hand, of the polymer brush composition, especially from the PNIPAAm content, and, on the other hand, of the peptide dimension and its reactivity.

pH and Salt Response of Mixed Brushes Made of Oppositely Charged Polyelectrolytes Studied by in Situ AFM Force Measurements and Imaging.

The response of mixed brushes made of poly(acrylic acid) and poly(2-vinyl pyridine) with a mixing ratio of about 60:40 was studied using atomic force microscopy (AFM) force measurements with colloidal probes and AFM imaging with a sharp tip in the pH range between 2.5 and 8 and at varying KCl concentrations up to 1 M. It was found that under all conditions a dense polyelectrolyte complex layer coexists with excess polyelectrolyte chains in varying swelling states depending on pH and salt concentration. The mixed brush thus combines typical features of polyelectrolyte brushes and complexes. So, the increase of the salt concentration not only led to a transition from osmotic to salted brush regime but also to salt-induced softening or partial decomposition of the complex layer. Attractive forces at high salt concentrations indicated the presence of P2VP chains in the swollen layer even at high pH values.

Salt Sensitivity of the Thermoresponsive Behavior of PNIPAAm Brushes.

We report investigations on the salt sensitivity of the thermoresponsive behavior of PNIPAAm brushes applying the quartz crystal microbalance coupled with spectroscopic ellipsometry technique. This approach enables a detailed study of the optical and mechanical behavior of the polymer coatings. Additional conclusions can be drawn from the difference between both techniques due to a difference in the contrast mechanism of both methods. A linear shift of the phase-transition temperature to lower temperatures with the addition of sodium chloride was found, similar to the behavior of free polymer chains in solution. The thermal hysteresis was found to be decreased by the addition of sodium chloride to the solution, hinting to the interaction of the ions with the amide groups of the polymer, whereby the formation of hydrogen bonds is hindered. The results of this study are of relevance to the application of PNIPAAm brushes in biological fluids and demonstrate the additional potential of the ion sensitivity besides the better known thermosensitivity.

Ligand displacement induced morphologies in block copolymer/quantum dot hybrids and formation of core-shell hybrid nanoobjects.

We investigate the self-assembly of a cylinder-forming polystyrene-block-poly(4-vinylpyridine) (PS-b-P4VP) block copolymer (BCP) mixed with trioctylphosphine oxide (TOPO) capped cadmium selenide (CdSe) quantum dots (QDs). The QDs were found to be enthalpically compatible with the P4VP chains via ligand displacement of TOPO from the QD surface. However, the QDs were found to localize preferentially at the PS/P4VP interphase plausibly to gain translational entropy in order to further lower the energetics of the self-assembled structure. Interestingly, the morphological transformation observed with increasing weight fraction of the QDs in the BCP/QD composite was driven by the migration of the displaced TOPO from the QD surface to the PS phase, effectively increasing its total volume fraction. Hence, the PS-b-P4VP BCP with PS as the minority block displayed lamellar morphology in its composite with QDs. Furthermore, the preferred localization of the QDs at the PS/P4VP interface led to the formation of a trilayer lamellar morphology which was deduced from the suppression of the primary scattering peak, relative to higher order peaks in the SAXS data. The morphological transformation was accompanied by a significant increase in the domain spacing due to excessive stretching of the longer P4VP chains of the asymmetric block copolymer. However, in the PS-b-P4VP/CdSe composites with P4VP as the minority block, cylindrical morphology was retained and the domain spacing decreased due to dominance of the co-surfactant effect as well as interfacial localization of CdSe QDs. We also demonstrate that these PS-b-P4VP/CdSe self-assembled hybrid materials could further be used to obtain isolated core-shell nanoobjects, such as nanofibers and nanosheets, containing CdSe QDs. The nanoobjects so obtained exhibited photoluminescence properties typical of CdSe quantum dots. These photoluminescent polymer nanoobjects could have potential applications in biological targeting and fluorescence labeling.

Tunable Fluorescence of a Semiconducting Polythiophene Positioned on DNA Origami.

A novel approach for the integration of π-conjugated polymers (CPs) into DNA-based nanostructures is presented. Using the controlled Kumada catalyst-transfer polycondensation, well-defined thiophene-based polymers with controllable molecular weight, specific end groups, and water-soluble oligoethylene glycol-based side chains were synthesized. The end groups were used for the easy but highly efficient click chemistry-based attachment of end-functionalized oligodeoxynucleotides (ODNs) with predesigned sequences. As demonstrated by surface plasmon resonance spectroscopy, the prepared block copolymers (BCPs), P3(EO)3T-b-ODN, comprising different ODN lengths and specific or repetitive sequences, undergo specific hybridization with complementary, thiol-functionalized ODNs immobilized on a gold surface. Furthermore, the site-specific attachment of the BCPs to DNA origami structures is studied. We demonstrate that a nanoscale object, that is, a single BCP with a single ODN handle, can be directed and bound to the DNA origami with reasonable yield, site-specificity, and high spatial density. On the basis of these results, we are able to demonstrate for the first time that optical properties of CP molecules densely immobilized on DNA origami can be locally fine-tuned by controlling the attractive π-π-stacking interactions between the CPs. In particular, we show that the fluorescence of the immobilized CP molecules can be significantly enhanced by surfactant-induced breakup of π-π-stacking interactions between the CP's backbones. Such molecular control over the emission intensity of the CPs can be valuable for the construction of sophisticated switchable nanophotonic devices and nanoscale biosensors.

Silica-supported Au@hollow-SiO2 particles with outstanding catalytic activity prepared via block copolymer template approach.

Catalytically active Au@hollow-SiO2 particles embedded in porous silica support (Au@hollow-SiO2@PSS) were prepared by using spherical micelles from poly(styrene)-block-poly(4-vinyl pyridine) block copolymer as a sacrificial template. Drastic increase of the shell porosity was observed after pyrolytic removal of polymeric template because the stretched poly(4-vinyl pyridine) chains interpenetrating with silica shell acted as an effective porogen. The embedding of Au@hollow-SiO2 particles in porous silica support prevented their fusion during pyrolysis. The catalytic activity of Au@hollow-SiO2@PSS was investigated using a model reaction of catalytic reduction of 4-nitrophenol and reductive degradation of Congo red azo-dye. Significantly, to the best of our knowledge, Au@hollow-SiO2@PSS catalyst shows the highest activity among analogous systems reported till now in literature. Such high activity was attributed to the presence of multiple pores within silica shell of Au@hollow-SiO2 particles and easy accessibility of reagents to the catalytically active sites of the ligand-free gold surface through the porous silica support.

In Situ Monitoring of Linear RGD-Peptide Bioconjugation with Nanoscale Polymer Brushes.

Bioinspired materials mimicking the native extracellular matrix environment are promising for biotechnological applications. Particularly, modular biosurface engineering based on the functionalization of stimuli-responsive polymer brushes with peptide sequences can be used for the development of smart surfaces with biomimetic cues. The key aspect of this study is the in situ monitoring and analytical verification of the biofunctionalization process on the basis of three complementary analytical techniques. In situ spectroscopic ellipsometry was used to quantify the amount of chemisorbed GRGDS at both the homopolymer poly(acrylic acid) (PAA) brush and the binary poly(N-isopropylacrylamide) (PNIPAAm)-PAA brushes, which was finally confirmed by an acidic hydrolysis combined with a subsequent reverse-phase high-performance liquid chromatography analysis. In situ attenuated total reflection-Fourier transform infrared spectroscopy provided a step-by-step detection of the biofunctionalization process so that an optimized protocol for the bioconjugation of GRGDS could be identified. The optimized protocol was used to create a temperature-responsive binary brush with a high amount of chemisorbed GRGDS, which is a promising candidate for the temperature-sensitive control of GRGDS presentation in further cell-instructive studies.

Adsorption of enzymes to stimuli-responsive polymer brushes: Influence of brush conformation on adsorbed amount and biocatalytic activity.

Polyelectrolyte brushes can be utilized to immobilize enzymes on macroscopic surfaces. This report investigates the influence of the pH value of the surrounding medium on the amount and the activity of enzymes adsorbed to poly(2-vinylpyridine) and poly(acrylic acid) brushes, as well as the creation of thermoresponsive biocatalytically active coatings via the adsorption of enzymes onto a mixed brush consisting of a polyelectrolyte and temperature-sensitive poly(N-isopropylacryl amide). Spectroscopic ellipsometry and attenuated total reflection-Fourier transform infrared spectroscopy are used to monitor the adsorption process. Additionally, infrared spectra are evaluated in terms of the secondary structure of the enzymes. Glucose oxidase is used as a model enzyme, where the enzymatic activity is measured after different adsorption conditions. Poly(acrylic acid) brushes generally adsorb larger amounts of enzyme, while less glucose oxidase is found on poly(2-vinylpyridine), which however exhibits higher specific activity. This difference in activity could be attributed to a difference in secondary structure of the adsorbed enzyme. For glucose oxidase adsorbed to mixed brushes, switching of enzymatic activity between an active state at 20°C and a less active state at 40°C as compared to the free enzyme in solution is observed. However, this switching is strongly depending on pH in mixed brushes of poly(acrylic acid) and poly(N-isopropylacryl amide) due to interactions between the polymers.

Swelling and Surface Interactions of End-Grafted Poly(2-vinylpyridine) Layers in Acidic Solution: Influence of Grafting Density and Salt Concentration.

In previous studies, the authors found that end-grafted layers of the weak polybase poly(2-vinylpyridine) (P2VP) in aqueous solutions do not only swell and collapse if the pH value and salt concentration are varied but also exhibit a pH- and salinity-dependent adhesion to microsized silica spheres. For a better understanding of these effects, in situ force measurements using the AFM colloidal probe technique were applied to end-grafted P2VP layers of different grafting densities in NaCl solutions at pH 2.5. Although a mushroom-to-brush transition could be seen in the dry state, the layers were in the brush regime in aqueous solutions at all NaCl concentrations and grafting densities. We observed an increase of the brush height with increasing grafting density and a salinity-dependent collapse and reswelling of the brushes. The adhesion between the P2VP layer and a silica sphere depended on both grafting density and salinity. At low salt concentrations, the adhesion reached its highest value at the intermediate grafting density and disappeared with denser brushes. Maximum adhesion was obtained for high NaCl concentrations and the lowest grafting density. From a detailed analysis of the experiments, we gained insight into chain stretching and density profiles under complex ionic conditions and into the mechanism of adhesion of polyelectrolytes to solid surfaces.

High Conductivity in Molecularly p-Doped Diketopyrrolopyrrole-Based Polymer: The Impact of a High Dopant Strength and Good Structural Order.

[3]-Radialene-based dopant CN6-CP studied herein, with its reduction potential of +0.8 versus Fc/Fc+ and the lowest unoccupied molecular orbital level of -5.87 eV, is the strongest molecular p-dopant reported in the open literature, so far. The efficient p-doping of the donor-acceptor dithienyl-diketopyrrolopyrrole-based copolymer having the highest unoccupied molecular orbital level of -5.49 eV is achieved. The doped films exhibit electrical conductivities up to 70 S cm(-1) .

Bienzymatic Sequential Reaction on Microgel Particles and Their Cofactor Dependent Applications.

We report, the preparation and characterization of bioconjugates, wherein enzymes pyruvate kinase (Pk) and l-lactic dehydrogenase (Ldh) were covalently bound to poly(N-isopropylacrylamide)-poly(ethylenimine) (PNIPAm-PEI) microgel support using glutaraldehyde (GA) as the cross-linker. The effects of different arrangements of enzymes on the microgels were investigated for the enzymatic behavior and to obtain maximum Pk-Ldh sequential reaction. The dual enzyme bioconjugates prepared by simultaneous addition of both the enzymes immobilized on the same microgel particles (PL), and PiLi, that is, dual enzyme bioconjugate obtained by combining single-enzyme bioconjugates (immobilized pyruvate kinase (Pi) and immobilized lactate dehydrogenase (Li)), were used to study the effect of the assembly of dual enzymes systems on the microgels. The kinetic parameters (Km, kcat), reaction parameters (temperature, pH), stability (thermal and storage), and cofactor dependent applications were studied for the dual enzymes conjugates. The kinetic results indicated an improved turn over number (kcat) for PL, while the kcat and catalytic efficiency was significantly decreased in case of PiLi. For cofactor dependent application, in which the ability of ADP monitoring and ATP synthesis by the conjugates were studied, the activity of PL was found to be nearly 2-fold better than that of PiLi. These results indicated that the influence of spacing between the enzymes is an important factor in optimization of multienzyme immobilization on the support.