Fluorescence study of the interaction between albumin and layered double hydroxides.

Layered double hydroxides nanoparticles (LDH-NP) are increasingly studied for biomedical applications. Nevertheless, their interaction with biomolecules such as proteins needs further exploration for an effective application. In this work, the adsorption of bovine serum albumin (BSA) on LDH-NP and the conformation changes of the protein upon adsorption were characterized using fluorescence spectroscopy. First, the quenching of tryptophan residues of BSA by chloride-intercalated LDH-NP was explored and the BSA adsorption capacity of LDH-NP were determined. Then, the structural conformation of the protein was analyzed by fluorescence spectroscopy (including synchronous, polarization and quenching studies) at different surface coverages. Finally, the proclivity of adsorbed BSA molecules to assemble as amyloid fibril was evaluated. Due to the positive charging and low curvature of LDH-NP, BSA molecules were strongly adsorbed, which produced a quenching of the protein fluorescence and a large adsorption capacity. The effect on BSA conformation was dependent on surface coverage (SC): at low values ,t he tryptophan residues were in more hydrophobic environments and more accessible to quenchers than al high ones. At low SC, there is space between the BSA molecules to spread on the surface, which led to a conformation change. Contrarily, the native conformation around tryptophan residues of BSA was preserved at high SC due to the tight packing of the adsorbed protein molecules. As a result, BSA molecules are stabilized against the formation of amyloid fibrils at high SC, while at low SC they present a similar fibrillation than free BSA.

A closer look into the physical interactions between lipid membranes and layered double hydroxide nanoparticles.

Layered double hydroxide nanoparticles (LDH-NPs) constitute promising nanocarriers for drug and gene delivery. Although their cell internalization has been studied, the interaction between LDH-NPs and biological membrane models, such as giant unilamellar vesicles (GUVs), remains unexplored. These vesicles are widely-used membrane models that allow minimizing the complexity and uncertainty associated with biological systems to study the physical interactions in the absence of cell metabolism effects. With such an approach the physicochemical properties of the membrane can be differentiated from the biological functionalities involved in cell internalization and the membrane-mediated internalization can be directly understood. In this work, we describe for the first time the interaction of LDH-NPs with freestanding negatively charged POPC:POPS GUVs by fluorescence microscopy. The experiments were performed with fluorescein labeled LDH-NPs of about 100 nm together with different fluorophores in order to evaluate the NPs interactions with the vesicles as well as their impact on the membrane morphology and permeability. Positively charged LDH-NPs are electrostatically accumulated at the GUVs membrane, altering its lateral phospholipid distribution and increasing the stiffness and permeability of the membrane. The adsorption of albumin (LDH@ALB) or polyacrylic acid (LDH@PA) passivates the surface of LDH-NPs eliminating long-range electrostatic attraction. The absence of membrane-mediated internalization of either LDH@ALB or LDH@PA, represents an advantage in the use of LDH-NPs as drug or nucleic acids nanocarriers, because suitable functionalization will allow an optimal cell targeting.

A simple surface biofunctionalization strategy to inhibit the biofilm formation by Staphylococcus aureus on solid substrates.

Staphylococcus aureus is an important opportunistic pathogen that causes a broad range of infections due to the bacteria capacity to form biofilms on medical devices. This work is aimed at inhibiting the biofilm formation by S. aureus on solid substrates using a simple surface biofunctionalization strategy. We previously found that surface biofunctionalization with structural perturbed albumin inhibited the initial stage of S. aureus adhesion. The current work extends this strategy with other plasma protein, fibrinogen, which in addition can be bond specifically to the cell wall-anchored proteins of S. aureus. The study of fibrinogen adsorption indicates that the fraction of surface-perturbed molecules is enlarged at long adsorption times and low protein concentration. In these conditions, a significant diminution of ca.60% of alive adhered bacteria were observed after 40 min and the biofilm formation was completely prevented. Thus, it seems that the inhibition of bacterial adhesion on substrates with surface-perturbed proteins represents a general trend even when specific interactions are present. On this basis, we developed a simple strategy to inhibit the formation of S. aureus biofilm, using thermally treated albumin or fibrinogen molecules prior to the substrate biofunctionalization. This strategy shows an excellent performance since the alive adhered bacteria diminishes ca. 90% at short incubation time, followed by the fully inhibition of biofilm formation. This novel and simple resource represents a change of the usual notion in avoiding post-surgery infections, mostly related to the use of medical devices.

Relevance of protein-protein interactions on the biological identity of nanoparticles.

Considering that the use of nanoparticles (NPs) as carriers of therapeutic or theranostic agents has increased in the last years, it is mandatory to understand the interaction between NPs and living systems. In contact with biological fluids, the NPs (synthetic identity) are covered with biomolecules that form a protein corona, which defines the biological identity. It is well known that the protein corona formation is mediated by non-specific physical interactions, but protein-protein interactions (PPI), involving specific recognition sites of the polypeptides, are also involved. This work explores the relationship between the synthetic and biological identities of layered double hydroxides nanoparticles (LDH-NPs) and the effect of the protein corona on the cellular response. With such a purpose, the synthetic identity was modified by coating LDH-NPs with either a single protein or a complex mixture of them, followed by the characterization of the protein corona formed in a commonly used cell culture medium. A proteomic approach was used to identify the protein corona molecules and the PPI network was constructed with a novel bioinformatic tool. The coating on LDH-NPs defines the biological identity in such a way that the composition of the protein corona as well as PPI are changed. Electrostatic interactions appear not to be the only driving force regulating the interactions between NPs, proteins and cells since the specific recognition also play a fundamental role. However, the biological identity of LDH-NPs does not affect the interactions with cells that shows negligible cytotoxicity and high internalization levels.

Albumin biofunctionalization to minimize the Staphylococcus aureus adhesion on solid substrates.

Staphylococcus aureus has become the most common opportunistic microorganism related to nosocomial infections due to the bacteria capacity to form biofilms on biomedical devices and implants. Since bacterial adhesion is the first step in this pathogenesis, it is evident that inhibiting such a process will reduce the opportunity for bacterial colonization on the devices. This work is aimed at optimizing a surface biofunctionalization strategy to inhibit the adhesion of S. aureus on solid substrates. The first part of the work deals with the albumin adsorption-desorption process, studied by a factorial design of experiments to explore a wide range of experimental factors (protein concentration, pH, flow rate and adsorption time) and responses (initial adsorption rate, adsorbed amount, desorbed extent) for hydrophilic and hydrophobic substrates, with a reduced number of experiments. This approach allows the simultaneous evaluation of the factors affecting the albumin adsorption-desorption process to find a qualitative correlation with the amount of alive S. aureus adhered on albumin biofunctionalized substrates. The results of this work point to a relationship between bacterial adhesion and the degree of albumin relaxation on the solid substrate. In fact, the inhibition of bacterial adhesion on albumin biofunctionalized substrates is due to the surface perturbation on the native structure of the protein. On this base, a biofunctionalization strategy was designed using a solution of thermally treated albumin molecules (higher ß-sheet or unordered secondary structure elements) to biofunctionalize solid substrates by dipping. With these albumin biofunctionalized substrates S. aureus adhesion was minimized.

Effect of the protein corona on the colloidal stability and reactivity of LDH-based nanocarriers.

The physicochemical properties of drug nanocarriers such as layered double hydroxide nanoparticles (LDH-NPs) determine their circulation times in biological media and their interaction with the targeted cells. Nevertheless, the components of the biological fluid, and particularly the formation of a protein corona, change the properties of as-prepared nanocarriers. Here, we discuss the effect of the protein corona formation on the colloidal stability and reactivity of LDH-NPs intercalated with chloride (LDH-Cl), carbonate (LDH-CO3) or dodecylsulfate (LDH-DS). These solids present model physicochemical properties (hydrophillic character, surface charge, and exchange capacity) that can be obtained depending on the interaction of drugs with LDH layers. The colloidal stability of LDH-NPs was determined in simulated biological fluids at high ionic strength and/or the presence of albumin (the main protein of human blood plasma), whereas the reactivity was evaluated by dissolution kinetics in acidic media, compatible with the environment of cell internalized nanocarriers. The protein corona increased the colloidal stability of the nanocarriers by steric hindrance at high ionic strength, reverted the positive zeta potential of as-prepared LDH-NPs and protected them from dissolution at low pHs. The properties of the anionic cargo of LDH-NPs strongly affected the protein corona and hence the fate of NPs in biological fluids. Drug nanocarriers with interfacial properties similar to those of LDH-Cl and LDH-CO3 seem to be more promising than LDH-DS in forming a protein corona. Then, LDH-Cl and LDH-CO3 would enable long circulation times due to their size, colloidal stability and low protein damage. Our results indicate that LDH-NPs preserve and even improve their properties as drug nanocarriers after interacting with the biological media, particularly their ability to reach the site of therapeutic action from the injection place.

Unaffected features of BSA stabilized Ag nanoparticles after storage and reconstitution in biological relevant media.

Silver-coated orthopedic implants and silver composite materials have been proposed to produce local biocidal activity at low dose to reduce post-surgery infection that remains one of the major contributions to the patient morbidity. This work presents the synthesis combined with the characterization, colloidal stability in biological relevant media, antimicrobial activity and handling properties of silver nanoparticles (Ag-NP) before and after freeze dry and storage. The nanomaterial was synthesized in aqueous solution with simple, reproducible and low-cost strategies using bovine serum albumin (BSA) as the stabilizing agent. Ag-NP were characterized by means of the size distribution and morphology (UV-vis spectra, dynamic light scattering measurements and TEM images), charge as a function of the pH (zeta potential measurements) and colloidal stability in biological relevant media (UV-vis spectra and dynamic light scattering measurements). Further, the interactions between the protein and Ag-NP were evaluated by surface enhanced Raman spectroscopy (SERS) and the antimicrobial activity was tested with two bacteria strains (namely Staphylococcus aureus and Staphylococcus epidermidis) mainly present in the infections caused by implants and prosthesis in orthopedic surgery. Finally, the Ag-NP dispersed in aqueous solution were dried and stored as long-lasting powders that were easily reconstituted without losing their stability and antimicrobial properties. The proposed methods to stabilize Ag-NP not only produce stable dispersions in media of biological relevance but also long-lasting powders with optimal antimicrobial activity in the nanomolar range. This level is much lower than the cytotoxicity determined in vitro on osteoblasts, osteoclasts and osteoarthritic chondrocytes. The synthesized Ag-NP can be incorporated as additive of biomaterials or pharmaceutical products to confer antimicrobial activity in a powdered form in different formulations, dispersed in aqueous and non-aqueous solutions or coated on the surface of different materials.

Surface coverage dictates the surface bio-activity of D-amino acid oxidase.

This work presents a systematic study on the relationship between the adsorption mechanism and the surface bio-activity of D-amino acid oxidase (pkDAAO). This rational approach is based on measuring the characteristic filling and relaxation times under different experimental conditions. With such a goal, real-time adsorption-desorption experiments at different degrees of surface coverage were performed tuning the electrostatic and hydrophobic interactions by changing the pH condition for the adsorption and the substrate properties (silica or gold). Surface bio-activity was measured in situ by amperometry using the bio-functional surface as the working electrode and ex situ by spectrophotometry. On both solid substrates, pkDAAO adsorption is a transport-controlled process, even under unfavorable electrostatic interactions (charged protein and substrate with the same sign) due to the high percentage of basic amino acids in the enzyme. On silica, the relaxation step is electrostatic in nature and occurs in the same time-scale as filling the surface when the substrate and the enzyme are oppositely charged at low surface coverage. Under unfavorable electrostatic conditions, the relaxation (if any) occurs at long time. Accordingly, the bio-activity of the native pkDAAO is preserved at any surface coverage. On gold, this step is driven by hydrophobic interactions (pH-independent) and the surface bio-activity is highly dependent on the degree of surface coverage. Under these conditions, the surface bio-activity is preserved only at high surfaces coverage. Our results clearly indicate that pkDAAO bio-functionalized surfaces cannot be coupled to amperometry because the analyte interferes the electrochemical signal. However, this simple bio-functionalized strategy can be joined to other detection methods.

Driving forces for the adsorption of a His-tag Chagas antigen. A rational approach to design bio-functional surfaces.

In order to rationally design a bio-functional surface based on the adsorption of a His-tag antigen, three requirements have to be considered: the bio-recognition element, the driving forces for the adsorption process and the detection mode of the bio-recognition event. This work is focused on the study of the adsorption mechanism of the His-tag H49 Chagas antigen on Ni(II) modified substrates. In order to construct the bio-functional surface, the gen of the H49 Chagas antigen was modified to incorporate His6 moiety at the N-terminal (His6-H49). Then, its physical adsorption and bio-affinity interaction with the solid substrate was studied by reflectometry. Besides His-Ni(II) bio-affinity interactions, His6-H49 was also physically adsorbed on Ni(II) modified substrates, leading to randomly oriented antigens. These loosely attached bio-molecules were partially removed using conditions of electrostatic repulsion. On the other hand, bio-affinity interactions, resulting in site-oriented molecules on the substrate, were only removable by specific competitors for Ni(II) surface sites. Finally, the surface bio-activity was determined from the peak separations of voltammetry waves due to the change of the electron transfer kinetics of a redox probe through the bio-functional surface (working electrode).

Ni(ii)-modified solid substrates as a platform to adsorb His-tag proteins.

This work investigated a simple and versatile modification to a solid substrate to develop electrochemical bio-recognition platforms based on bio-affinity interactions between histidine (His)-tagged proteins and Ni(ii) surface sites. Carboxylate (COO)-functionalized substrates were prepared in multiple steps, initiated with an amino-terminated self-assembled monolayer (SAM) on polycrystalline gold. Surface enhanced Raman spectroscopy (SERS), quartz crystal microbalance with dissipation monitoring (QCM-D) and contact angle measurements were used to follow the modification process. Upon completion of the modification process, the surface COO-Ni(ii) chelate complex and the coordination mode used to bind the His-tag proteins were characterized by X-ray absorption near-edge spectroscopy (XANES). Finally, the electrochemical stability and response of the modified substrates were evaluated. The versatility of the modification process was verified using silica as the substrate. QCM-D and SERS results indicated that two types of films were formed: a COO-terminated SAM, which resulted from the reduction of previously incorporated surface aldehyde groups, and a physically adsorbed polymeric glutaraldehyde film, which was produced in the alkaline medium. XANES spectral features indicated that COO-Ni(ii) formed a non-distorted octahedral complex on the substrate. The electrochemical stability and response towards a redox mediator of the COO-Ni(ii)-terminated SAM indicated that this platform could be easily coupled to an electrochemical method to detect bio-recognition events.

The effect of interlayer anion on the reactivity of Mg-Al layered double hydroxides: improving and extending the customization capacity of anionic clays.

Layered double hydroxides (LDHs) reactivity and interfacial behavior are closely interconnected and control particle properties relevant to the wide range of these solids' applications. Despite their importance, their relationship has been hardly described. In this work, chloride and dodecylsulfate (DDS(-)) intercalated LDHs are studied combining experimental data (electrophoretic mobility and contact angle measurements, hydroxyl and organic compounds uptake) and a simple mathematical model that includes anion-binding and acid-base reactions. This approach evidences the anion effect on LDHs interfacial behavior, reflected in the opposite particle charge and the different surface hydrophobic/hydrophilic character. LDHs reactivity are also determined by the interlayer composition, as demonstrated by the cation uptake capability of the DDS(-) intercalated sample. Consequently, the interlayer anion modifies the LDHs interfacial properties and reactivity, which in turn extends the customization capacity of these solids.

Infrared study of trifluoroacetic acid unpurified synthetic peptides in aqueous solution: trifluoroacetic acid removal and band assignment.

Synthetic peptide or protein samples are mostly unpurified with trifluoroacetic acid (TFA) used during the synthesis procedure, which strongly interferes with structure determination by infrared (IR) spectroscopy. The aim of this work was to propose a simple strategy to remove TFA contribution from attenuated total reflection (ATR)-IR spectra of the hexahistidine peptide (His6) in aqueous solution to study the conformation of this synthetic peptide without previous purification. Such a strategy is based on the subtraction mode widely employed to remove water contribution, and it is tested with TFA unpurified histidine as a model system. The subtraction is based on eliminating the strong TFA bands at 1147 and 1200cm(-1) by applying a scaling factor (as in buffer correction). The proposed modes represent excellent strategies that do not modify spectral features, and they provide reliable routines to obtain the synthetic peptide spectrum without TFA contribution. The conformational information from the corrected spectra at different pH values is deduced from semiempirical calculated IR spectra of different His6 conformers. The spectral features and the band positions of the corrected spectrum suggest that the peptide molecules mainly adopt an intermolecular ß-sheet structure.

Bio-recognition capability of Streptomyces sp. M7 evaluated in adverse conditions for use as a biological transducer in a Lindane biosensor.

Bio-recognition devices have captured special attention because they combine biological specificity and selectivity with electronics to perform environmental and biomedical analysis. Lindane is a recalcitrant pesticide considered potential carcinogen that has caused serious pollution problems. The purpose of the present study is to evaluate Streptomyces sp. M7 ability to dechlorinate lindane in liquid defined media in adverse culture conditions. Bacterial activity was monitored by electrochemical impedance spectroscopy. These results confirm that the microorganism adhered to a solid support is able to grow and to metabolize the organochlorine pesticide as a sole carbon source. Therefore, Streptomyces sp. M7 can be applied for a future development of a prototype for lindane detection and quantification.

Dissolution kinetics and mechanism of Mg-Al layered double hydroxides: a simple approach to describe drug release in acid media.

Layered double hydroxides (LDHs) weathering in acidic media is one of the main features that affects their applications in drug delivery systems. In this work, the dissolution kinetics of biocompatible Mg-Al LDHs was studied at different initial pH values and solid concentrations using a simple and fast experimental method that coupled flow injection analysis and amperometric detection. A carbonate intercalated sample was used to determine the controlling step of the process and the dissolution mechanism. Finally, the study was extended to an ibuprofen intercalated LDH. The obtained results showed that the weathering process was mainly controlled by the exposed area and surface reactivity of LDHs particles. The dissolution mechanism at the particle surface was described in two steps: fast formation of surface reactive sites by hydroxyl group protonation and slow detachment of metal ions from surface. At strongly acidic conditions, the reaction rate was pH dependent due to the equilibrium between protonated (active) and deprotonated (inactive) hydroxyl groups. On the other hand, at mildly acidic conditions, the dissolution behavior was also ruled by the equilibrium attained between the particle surface reactive sites and the dissolved species. LDHs solubility and dissolution rate presented strong dependence with the interlayer anion. The ibuprofen intercalated sample was more soluble and more rapidly dissolved than the carbonate intercalated one in acetic/acetate buffer. On the other hand, the dissolution mechanism was invariant with the interlayer anion.

Electrostatic and hydrophobic interactions involved in CNT biofunctionalization with short ss-DNA.

This work is aimed at studying the adsorption mechanism of short chain 20-mer pyrimidinic homo-ss-DNA (oligodeoxyribonucleotide, ODN: polyC(20) and polyT(20)) onto CNT by reflectometry. To analyze the experimental data, the effective-medium theory using the Bruggemann approximation represents a suitable optical model to account for the surface properties (roughness, thickness and optical constants) and the size of the adsorbate. Systematic information about the involved interactions is obtained by changing the physico-chemical properties of the system. Hydrophobic and electrostatic interactions are evaluated by comparing the adsorption on hydrophobic CNT and on hydrophilic silica and by modulating the ionic strength with and without Mg(2+). The ODN adsorption process on CNT is driven by hydrophobic interactions only when the electrostatic repulsion is suppressed. The adsorption mode results in ODN molecules in a side-on orientation with the bases (non-polar region) towards the surface. This unfavorable orientation is partially reverse by adding Mg(2+). On the other hand, the adsorption on silica is dominated by the strong repulsive electrostatic interaction that is screened at high ionic strength or mediated by Mg(2+). The cation-mediated process induces the interaction of the phosphate backbone (polar region) with the surface, leaving the bases free for hybridization. Although the general adsorption behavior of the pyrimidine bases is the same, polyC(20) presents higher affinity for the CNT surface due to its acid-base properties.

Determination of a setup correction function to obtain adsorption kinetic data at stagnation point flow conditions.

This paper is the first report on the characterization of the hydrodynamic conditions in a flow cell designed to study adsorption processes by spectroscopic ellipsometry. The resulting cell enables combining the advantages of in situ spectroscopic ellipsometry with stagnation point flow conditions. An additional advantage is that the proposed cell features a fixed position of the "inlet tube" with respect to the substrate, thus facilitating the alignment of multiple substrates. Theoretical calculations were performed by computational fluid dynamics and compared with experimental data (adsorption kinetics) obtained for the adsorption of polyethylene glycol to silica under a variety of experimental conditions. Additionally, a simple methodology to correct experimental data for errors associated with the size of the measured spot and for variations of mass transfer in the vicinity of the stagnation point is herein introduced. The proposed correction method would allow researchers to reasonably estimate the adsorption kinetics at the stagnation point and quantitatively compare their results, even when using different experimental setups. The applicability of the proposed correction function was verified by evaluating the kinetics of protein adsorption under different experimental conditions.

Interaction of D-amino acid oxidase with carbon nanotubes: implications in the design of biosensors.

We have investigated the interaction of d-amino acid oxidase (DAAO) with single-walled carbon nanotubes (CNT) by spectroscopic ellipsometry. Dynamic adsorption experiments were performed at different experimental conditions. In addition, the activity of the enzyme adsorbed at different conditions was studied. Our results indicate that DAAO can be adsorbed to CNT at different pH values and concentrations by a combination of hydrophobic and electrostatic interactions. Considering that the highest enzymatic activity was obtained by adsorbing the protein at pH 5.7 and 0.1 mg x mL(-1), our results indicate that DAAO can adopt multiple orientations on the surface, which are ultimately responsible for significant differences in catalytic activity.

EDTA modified LDHs as Cu2+ scavengers: removal kinetics and sorbent stability.

EDTA modified layered double hydroxides (LDHs) were investigated as potential sorbents to remediate heavy metals pollution. The polidentate ligand was introduced by an exchange method in a Zn-Al-LDH, which takes place with partial erosion of the layers, causing the intercalation of [Zn(EDTA)](2-) complex instead of the ligand. [Cu(H(2)O)(6)](2+) cation was selected as a model cation to study the uptake mechanism, exploring the elimination kinetics from the first minutes up to the steady state. A flow injection analysis system coupled to an amperometric detector (FIA-AM) was applied to perform fast and reliable [Cu(H(2)O)(6)](2+) determinations in monodisperse solid-aqueous solution systems. Furthermore, the sorbent stability was determined as a function of the pH and the nitrate concentration. The [Cu(H(2)O)(6)](2+) elimination is produced by an exchange reaction with [Zn(EDTA)](2-) anions placed either in the solid interlayer or in the aqueous solution, this last being released from the sorbent. Additional [Cu(H(2)O)(6)](2+) removal is produced by Cu(OH)(2) precipitation at high copper concentrations due to the LDHs high pH buffering capacity. The sorbent removes [Cu(H(2)O)(6)](2+) with high affinity in a wide concentration range. The elimination process reaches equilibrium in less than 30 min and leaves metal cation concentrations lower than 0.05 ppm in the supernatants.

Latex of immunodiagnosis for detecting the Chagas disease: II. Chemical coupling of antigen Ag36 onto carboxylated latexes.

A novel immunodiagnosis reagent for detecting the Chagas Disease was developed, by chemical coupling of antigen Ag36 of Trypanosoma cruzi onto two (carboxylated and core-shell) latexes. The coupling reactions involved the use of a carbodiimide intermediate. Bovine serum albumin (BSA) was used as a model protein for determining the appropriate conditions for its physical and chemical coupling. BSA showed an increased adsorption onto the base carboxylated latexes, with respect to a PS latex without carboxyl groups. The chemical bonding experiments only involved the carboxylated latexes. With BSA, the final density of covalently bound protein was 2.30 mg/m(2). In addition, around 55% of the total linked protein was chemically coupled, and the reaction was little affected by the pH. With Ag36, the final density of covalently bound protein was 2.44 mg/m(2), around 80% of the total linked protein was chemically coupled, and the chemical coupling was maximum at pH = 5 (i.e., close to the isoelectric point).

Electrophoretic effects of the adsorption of anionic surfactants to poly(dimethylsiloxane)-coated capillaries.

Poly(dimethylsiloxane) (PDMS) is one of the most convenient materials to construct capillary electrophoresis microchips. Even though PDMS has many advantages, its use is often limited by its hydrophobicity. Although it is well-known that the surface properties of PDMS can be modified by anionic surfactants, very little is known regarding the driving forces or the electrophoretic consequences of the adsorption of anionic surfactants. In this work, the adsorption of alkyl surfactants on PDMS was studied by performing electroosmotic flow (microEOF) measurements. In order to mimic the behavior of PDMS microchannels, fused-silica capillaries were coated with PDMS and used for the microEOF measurements. This approach allowed using standard CE instrumentation and provided significant advantages over similar experiments performed on microchips. The change in the microEOF in the presence of surfactants was correlated to the surfactant adsorbed amount which, plotted versus surfactant concentration, gives an adsorption isotherm. The adsorption isotherms were obtained using alkyl surfactants with different chain lengths and head groups. According to our results, the interaction of alkyl surfactants with the PDMS surface is determined by a combination of hydrophobic and electrostatic interactions, where the former is more significant than the latter. The affinity of each surfactant for the PDMS surface was calculated by fitting the adsorption profiles with a Langmuir equation and, in the case of single-charged surfactants, correlated to the corresponding cmc value.