Gotowa bibliografia na temat „Surfactant-Protein System - Biomolecular Devices”

The fabrication of efficient organic electrochemical transistors (OECTs)-based biosensors requires the design of biocompatible interfaces for the immobilization of biorecognition elements, as well as the development of robust channel materials to enable the transduction of the biochemical event into a reliable electrical signal. In this work, PEDOT-polyamine blends are shown as versatile organic films that can act as both highly conducting channels of the transistors and non-denaturing platforms for the construction of the biomolecular architectures that operate as sensing surfaces. To achieve this goal, we synthesized and characterized films of PEDOT and polyallylamine hydrochloride (PAH) and employed them as conducting channels in the construction of OECTs. Next, we studied the response of the obtained devices to protein adsorption, using glucose oxidase (GOx) as a model system, through two different strategies: The direct electrostatic adsorption of GOx on the PEDOT-PAH film and the specific recognition of the protein by a lectin attached to the surface. Firstly, we used surface plasmon resonance to monitor the adsorption of the proteins and the stability of the assemblies on PEDOT-PAH films. Then, we monitored the same processes with the OECT showing the capability of the device to perform the detection of the protein binding process in real time. In addition, the sensing mechanisms enabling the monitoring of the adsorption process with the OECTs for the two strategies are discussed.

Silicon nanowire (SiNW) field-effect transistors (FETs) have been developed as very sensitive and label-free biomolecular sensors. The detection principle operating in a SiNW biosensor is indirect. The biomolecules are detected by measuring the changes in the current through the transistor. Those changes are produced by the electrical field created by the biomolecule. Here, we have combined nanolithography, chemical functionalization, electrical measurements and molecular recognition methods to correlate the current measured by the SiNW transistor with the presence of specific molecular recognition events on the surface of the SiNW. Oxidation scanning probe lithography (o-SPL) was applied to fabricate sub-12 nm SiNW field-effect transistors. The devices were applied to detect very small concentrations of proteins (500 pM). Atomic force microscopy (AFM) single-molecule force spectroscopy (SMFS) experiments allowed the identification of the protein adsorption sites on the surface of the nanowire. We detected specific interactions between the biotin-functionalized AFM tip and individual avidin molecules adsorbed to the SiNW. The measurements confirmed that electrical current changes measured by the device were associated with the deposition of avidin molecules.

Carbon nanotube field-effect biosensors (CNT-bioFETs) are ultraminiaturized devices that can be used to measure single-molecule kinetics of biomolecules on time scales going from a few microseconds to several minutes, as demonstrated for nucleic acid hybridization [1] and folding [2] as well as for enzyme function [3]. Experiments indicate that the sensitivity of CNT-bioFETs originates from the interplay between the nanotube’s conductance, which is monitored by the device, and the electrostatic potential generated by the biomolecule under investigation, which is localized on the nanotube [4,5]. The measured conductance exhibits characteristic transitions between two levels (or more) as a function of time, as the biomolecule folds or performs its function. Yet, the origin of this electrostatic gating of the carbon nanotube by a single biomolecule is not well understood at the molecular scale. To bridge this gap, we employ molecular dynamics (MD) and Hamiltonian replica exchange (HREX) simulations to unveil: (1) the interactions between the biomolecule and the nanotube to which it is attached in the device and (2) the electrostatic potential on the nanotube as the state of the biomolecule changes. We address these questions by considering three prototypical cases: the function of the Lysozyme protein, the hybridization of 10-nt DNA sequence and the folding of a DNA G-quadruplex, which were previously characterized using CNT-bioFETs [1-5]. Our simulations show that the lysozyme, the 10-nt DNA sequence and the DNA G-quadruplex interact differently with the nanotube to which they are attached. Consequently, the electrostatic potential (ESP) that they generate on the nanotube is very sensitive to the type and state of the biomolecule. When compared to experiment, the ESP distribution for the with-ligand and without-ligand states of the Lysozyme protein are in line with the measured two-level conductance by CNT-bioFETs. For the DNAs, however, the ESP distribution for their different states does not agree with the measured two-level conductance. Experiments imply that the DNA strand is not interacting with the nanotube, which is not what our simulations suggest. The reason for this apparent conflict could arise from the impact of the external electric field imposed by the gate electrode in CNT-bioFETs on highly charged systems such as DNAs, as supported by our recent simulations. The significance of this work is twofold. First, it contributes to a better understanding of the inner working of carbon nanotube field-effect biosensors, which is crucially needed to support the development of these promising devices in the lab. Second, it provides the structural ensemble of the biomolecules and their interactions with the nanotube in these devices, which can serve as a starting point for a finer characterization of their effect on the carbon nanotube’s conductance at the ab initio level. [1] S. Sorgenfrei et al. Nat Nanotechnol, 2011, 6, 126-132. [2] D. Bouilly et al. Nano Lett, 2016, 16, 4679-4685. [3] Y. Choi et al. Science, 2012, 335, 319-324. [4] S. Sorgenfrei et al. Nano Lett, 2011, 11, 3739-3743. [5] Y. Choi et al. Nano Lett, 2013, 13, 625-631.

Surface plasmon resonance (SPR) and Love wave (LW) surface acoustic wave (SAW) sensors have been established as reliable biosensing technologies for label-free, real-time monitoring of biomolecular interactions. This work reports the development of a combined SPR/LW-SAW platform to facilitate simultaneous optical and acoustic measurements for the investigation of biomolecules binding on a single surface. The system’s output provides recordings of two acoustic parameters, phase and amplitude of a Love wave, synchronized with SPR readings. We present the design and manufacturing of a novel experimental set-up employing, in addition to the SPR/LW-SAW device, a 3D-printed plastic holder combined with a PDMS microfluidic cell so that the platform can be used in a flow-through mode. The system was evaluated in a systematic study of the optical and acoustic responses for different surface perturbations, i.e., rigid mass loading (Au deposition), pure viscous loading (glycerol and sucrose solutions) and protein adsorption (BSA). Our results provide the theoretical and experimental basis for future application of the combined system to other biochemical and biophysical studies.

A variety of DNA, protein or cell microarray devices and systems have been developed and commercialized. In addition to the biomolecule related analysis, they are also being used for pharmacogenomic research, infectious and genetic disease and cancer diagnostics, and proteomic and cellular analysis.1 Currently, microarray is fabricated on a planar surface; this limits the amount of biomolecules that can be bounded on the surface. In this work, a planar protein microarray chip with nonplanar spot surface was fabricated to enhance the chip performance. A nonplanar spot surface was created by first coating the silica nanoparticles with albumin and depositing them into the patterned microwells. The curve surfaces of the nanoparticles increase the surface area for immobilization of proteins, which helps to enhance the detection sensitivity of the chip. Using this technique, proteins are immobilized onto the nanoparticles before they are deposited onto the chip, and therefore the method of protein immobilization can be customized at each spot. Furthermore, a nonplanar surface promotes the retention of native protein structure better than planar surface.2 The technique developed can be used to produce different types of microarrays, such as DNA, protein and antibody microarrays.

Purpose The purpose of this paper is to present possibility of fast and certain identification of bovine serum albumin (BSA) by means of ion-sensitive field effect transistor (ISFET) structures. Because BSA can cause allergic reactions in humans, it is one of reasons for development of sensitive sensors to detect residual BSA. BSA is commonly used in biochemistry and molecular biology in laboratory experiments. Therefore, to better understand the mechanism of signal transduction in simulated biological environment and to elucidate the role of adsorption of biomolecules in the generation of a signal at the interface with biological systems, the measurements of ISFET current response in the presence of BSA as a reference protein molecule were performed. Design/methodology/approach To fabricate transistors, silicon technology was used. The ISFET structures were coupled to specially designed double-side printed circuit board holder. After modification of the field effect transistor (FET) device with 3-aminopropyltriethoxysilane (APTES), a sensor with high sensitivity toward reference biomolecules was obtained. The current–voltage (I-V) characteristics of structures with and without gate modification were measured. Keithley SMU 236/237/238 measurement set was used. Deionized water solution and 0.05 per cent BSA were used. Findings In this research, a method of preparation of a biosensor based on a FET was developed. Sensitivity of APTES-modified FET device toward BSA as a biomolecule was investigated. I-V relationships of FET devices (with and without modification), being the effect of the interactions with the solution containing 0.05 per cent BSA, were measured and compared to the measurements performed for solutions without BSA. Originality value Compared to SiO2-containing ISFETs without modification or other different dielectrics, the application of APTES as the part of the membrane induced significant increase in their sensitivity to BSA.

Paper-based analytical devices have been substantially developed in recent decades. Many fabrication techniques for paper-based analytical devices have been demonstrated and reported. Herein, we report a relatively rapid, simple, and inexpensive method for fabricating paper-based analytical devices using parafilm hot pressing. We studied and optimized the effect of the key fabrication parameters, namely pressure, temperature, and pressing time. We discerned the optimal conditions, including a pressure of 3.8 MPa, temperature of 80 °C, and 3 min of pressing time, with the smallest hydrophobic barrier size (821 µm) being governed by laminate mask and parafilm dispersal from pressure and heat. Physical and biochemical properties were evaluated to substantiate the paper functionality for analytical devices. The wicking speed in the fabricated paper strips was slightly lower than that of non-processed paper, resulting from a reduced paper pore size after hot pressing. A colorimetric immunological assay was performed to demonstrate the protein binding capacity of the paper-based device after exposure to pressure and heat from the fabrication. Moreover, mixing in a two-dimensional paper-based device and flowing in a three-dimensional counterpart were thoroughly investigated, demonstrating that the paper devices from this fabrication process are potentially applicable as analytical devices for biomolecule detection. Fast, easy, and inexpensive parafilm hot press fabrication presents an opportunity for researchers to develop paper-based analytical devices in resource-limited environments.

We utilized electrophoresis to control the fluidity of sample biomolecules in sample aqueous solutions inside the nanochannel for single-molecule detection by using a nanochannel-integrated nanogap electrode, which is composed of a nano-gap sensing electrode, nanochannel, and tapered focusing channel. In order to suppress electro-osmotic flow and thermal convection inside this nanochannel, we optimized the reduction ratios of the tapered focusing channel, and the ratio of inlet 10 μm to outlet 0.5 μm was found to be high performance of electrophoresis with lower concentration of 0.05 × TBE (Tris/Borate/EDTA) buffer containing a surfactant of 0.1 w/v% polyvinylpyrrolidone (PVP). Under the optimized conditions, single-molecule electrical measurement of deoxyguanosine monophosphate (dGMP) was performed and it was found that the throughput was significantly improved by nearly an order of magnitude compared to that without electrophoresis. In addition, it was also found that the long-duration signals that could interfere with discrimination were significantly reduced. This is because the strong electrophoresis flow inside the nanochannels prevents the molecules’ adsorption near the electrodes. This single-molecule electrical measurement with nanochannel-integrated nano-gap electrodes by electrophoresis significantly improved the throughput of signal detection and identification accuracy.

Proteins on biomicroelectromechanical systems (BioMEMS) confer specific molecular functionalities. In planar FET sensors (field-effect transistors, a class of devices whose protein-sensing capabilities we demonstrated in physiological buffers), interfacial proteins are analyte receptors, determining sensor molecular recognition specificity. Receptors are bound to the FET through a polymeric interface, and gross disruption of interfaces that removes a large percentage of receptors or inactivates large fractions of them diminishes sensor sensitivity. Sensitivity is also determined by the distance between the bound analyte and the semiconductor. Consequently, differential properties of surface polymers are design parameters for FET sensors. We compare thickness, surface roughness, adhesion, friction and wear properties of silane polymer layers bound to oxides (SiO 2 and Al 2 O 3 , as on AlGaN HFETs). We compare those properties of the film–substrate pairs after an additional deposition of biotin and streptavidin. Adhesion between protein and device and interfacial friction properties affect FET reliability because these parameters affect wear resistance of interfaces to abrasive insult in vivo . Adhesion/friction determines the extent of stickage between the interface and tissue and interfacial resistance to mechanical damage. We document systematic, consistent differences in thickness and wear resistance of silane films that can be correlated with film chemistry and deposition procedures, providing guidance for rational interfacial design for planar AlGaN HFET sensors.

Electrospinning uses an electric field to produce fine fibers of nano and micron scale diameters from polymer solutions. Despite innovation in jet initiation, jet path control and fiber collection, it is common to only fabricate planar and tubular-shaped electrospun products. For applications that encapsulate cells and tissues inside a porous container, it is useful to develop biocompatible hollow core-containing devices. To this end, by introducing a 3D-printed framework containing a sodium chloride pellet (sacrificial core) as the collector and through post-electrospinning dissolution of the sacrificial core, we demonstrate that hollow core containing polyamide 66 (nylon 66) devices can be easily fabricated for use as cell encapsulation systems. ATR-FTIR and TG/DTA studies were used to verify that the bulk properties of the electrospun device were not altered by contact with the salt pellet during fiber collection. Protein diffusion investigations demonstrated that the capsule allowed free diffusion of model biomolecules (insulin, albumin and Ig G). Cell encapsulation studies with model cell types (fibroblasts and lymphocytes) revealed that the capsule supports the viability of encapsulated cells inside the capsule whilst compartmentalizing immune cells outside of the capsule. Taken together, the use of a salt pellet as a sacrificial core within a 3D printed framework to support fiber collection, as well as the ability to easily remove this core using aqueous dissolution, results in a biocompatible device that can be tailored for use in cell and tissue encapsulation applications.

Physically encapsulated droplet-interface bilayers are formed by confining aqueous droplets surrounded by lipid mono-layers in connected compartments within a solid substrate. The droplets reside within each compartment and are positioned on fixed electrodes built into the solid substrate. Full encapsulation of the network is achieved with a solid cap that inserts into the substrate to form a closed volume. Encapsulated networks provide increased portability over unencapsulated networks by limiting droplet movement and by integrating the electrodes into the supporting fixture. The formation of encapsulated droplet-interface bilayers is confirmed with electrical impedance spectroscopy and cyclic voltammetry is also used to measure the effect of alamethicin proteins incorporated into the resulting lipid bilayers. The durability of the networks is quantified using a mechanical shaker to oscillate the bilayer in a direction transverse to the plane of the membrane and the results show that single droplet-interface bilayers can withstand several g’s of acceleration. Observed failure modes include both droplet separation and bilayer rupturing, where the geometry of the supporting substrate and the presence of electrodes are key contributors. Physically encapsulated DIBs can be shaken, moved, and inverted without bilayer failure, enabling the creation of portable, protein-powered devices.

In this article, microdroplet motion in the electrocapillary-based digital microfluidic systems is modeled accurately, and the combined effects of the biomolecular adsorption and micro-droplet evaporation on the performance of the device are investigated. An electrohydrodynamic approach is used to model the driving and resisting forces, and Fick’s law and Gibbs equation are used to calculate the microdroplet evaporation and adsorption rate. Effects of the adsorption and evaporation rates are then implemented into the microdroplet dynamics by adding new terms into the force balance equation. It is shown that mass loss due to the evaporation tends to increase the protein concentration, and on the other hand, the increased concentration due to the mass loss increases the biomolecular adsorption rate which has a reverse effect on the concentration. The modeling results indicate that evaporation and adsorption play crucial roles in the microdroplet dynamics.

Microdialysis is a commonly used technique for separating small biomolecules within a complex biological mixture for continuous biochemical monitoring. Microdialysis is based upon controlling the mass transfer rate of small biomolecules diffusing across a semipermeable membrane into a dialysis fluid while excluding larger molecules such as proteins. These small molecules are subsequently sensed using a biosensor. Since many biosensors are extremely susceptible to fouling, their stability and lifetime can be extended if metabolites are filtered through a microdialysis membrane before the dialysis fluid is moved into the sensor. Dialysis is also used commonly in biological laboratories to desalt high ionic strength protein solutions. As biochemical analysis systems become more integrated for μTAS systems there is a need to automate this process. Thus, an on-chip dialysis system is useful for biochemical reaction engineering where very tight control of ionic conditions must be maintained for effective enzymatic activity. This work demonstrates the ability to integrate polymer microdialysis membranes with microfluidic systems. Microchannels are bonded with a regenerated cellulose membrane. After microchannels are produced using standard processing techniques, they are integrated with these membranes. The cellulose is activated in an oxygen plasma followed by a lamination bond to the microchannels at moderate pressure and elevated temperature. Devices were placed in a solution of rhodamine dye, and dialysis fluid was allowed to flow through the microchannels. The outlet dye concentration was measured by fluorescence intensity as a function of flow rate and follows analytically predicted results.

The outstanding characteristics of polydimethylsiloxane (PDMS) caused its extensive use as base material to manufacture microfluidic devices. PDMS has numerous advantages coming from instinct properties such as its low cost, simple fabrication procedure, and robust nature that make it a compatible material in many applications such as biological and biomedical engineering. In spite of favorable physical and chemical properties, hydrophobic surface of PDMS is sometimes debatable. Because of PDMS is highly hydrophobic, pumping aqueous solution through microchannels using only capillary forces might be difficult. Although many surface treatments methods have been proposed to modify and increase the hydrophilicity of PDMS [Oxygen plasma [1], UV-radiation [2], Silanization and Chemical vapour deposition [3],…], the use of surfactants is the most effective and easiest method to overcome the hydrophobicity compared to more complex protocols which require expensive facilities [4,5]. The hydrophilic behavior of surfactant-added PDMS and especially its biocompatibility has allowed many microfluidic bio-applications such as separation of biomolecules [6,7], blood cell separation [8] and cell-based assay [9,10]. This paper discusses about the efficiency of adding different surfactants on the wettability of PDMS.

A poly(dimethylsiloxane) (PDMS) microfluidic chip-based cartridge was fabricated by sandwiching commercial dialysis membrane and inserting fused-silica capillary into the end of channel according to the principle and structure of a commercial fused-silicon capillary-based cartridge, which can adapt to an IEF analyzer for isoelectric focusing with whole channel imaging detection (IEF-WCID). The novel design of sandwiching membrane in this chip not only eliminated the unfavorable hydrodynamic pressure, leading to poor IEF reproducibility, but made the sample injection much easy. Thus the reproducibility of analysis was very good. The prepared microfluidic chips were applied for qualitative and quantitative analysis of proteins. The six pI markers in the range of 3–10 were separated by IEF under the optimized conditions. The pH gradients exhibited good linear by plotting the pI versus peak position, and the correlation coefficient reached to 0.9994 and 0.9995. The separation of more complicated human hemoglobin control and myoglobin sample could be achieved. By comparison with the separation efficiency obtained on the microfluidic chip and commercial cartridge, the results were similar, which indicates the capillary cartridge may be replaced with the cost-efficient PDMS microfluidic chip. It is anticipated the high throughput analysis can be easily performed on this microfluidic chip patterned multi-channels. The techniques of capillary electrophoresis (CE) have been extensively explored for the chip-based separation. Isoelectric focusing (IEF) as one of high-resolution CE techniques has been widely applied for the separation of zwitterionic biomolecules, such as proteins and peptides. After the samples were focused at their corresponding pIs, the focused zones were mobilized to pass through the detection point for obtaining an electropherogram. This single-point detection imposes extensive restriction for chip-based IEF because a mobilization process requires additional time and lowers resolution and reproducibility of the separation [1]. An alternative is whole-column imaging detection developed by Pawliszyn et al [2] is an ideal detection method for IEF because no mobilization is required, which avoids the disadvantages as mentioned previously. Most microfluidic systems could be fabricated in glass/silicon or polymers in which the channels are defined using photolithography and micromachining. Mao and Pawliszyn [3] have developed a method for IEF on an etched quartz chip following whole-channel imaging detection (WCID). Ren et al [4] presented an integrated WCID system on glass microfluidic chip. However, these materials have some disadvantages such as expensive and fragile and so on. An attractive alternative for fabrication of microfluidic devices is using poly(dimethylsiloxane) (PDMS) as material, which has unique properties such as nontoxic, optical transparent down to 280 nm, elastomeric, hydrophobic surface chemistry Yao et al. [5] designed the glass/PDMS microchip integrated whole-column fluorescence imaging detection for IEF of R-phycoerythrin. Our preliminary studies have successfully developed a PDMS chip-based cartridge for IEF-WCID. It is due to hydrodynamic flow between two reservoirs that the focused zones were mobilized, thus gave poor reproducibility and difficulty in sample infusion. As membranes have been integrated into microchips for microdialysis, protein digestion, solid-phase extraction, desalting, pumping and so on, it could minimize hydrodynamic flow by using membranes as a filter. Although a simple PDMS chip-based cartridge has been successfully fabricated in our labs according to the principle of commercial capillary-based cartridge, it is difficult to introduce the sample into channel for IEF-WCID. As the vacuum was applied in one end of channel for infusing of solution into channel, the lifetime of this chip-based cartridge is shortened. Additionally, the hydrodynamic flow is occurred due to the different heights of anolyte and catholyte in two reservoirs, respectively. The IEF separation was deteriorated by the infusion of anolyte or catholyte, thus leading to poor reproducibility of IEF-WCID analysis. Similar to the hollow fiber in the commercial capillary-based cartridge in which it is aimed to separate the sample in the capillary and electrolytes in the reservoirs, porous membrane was integrated into PDMS chips for decrease of hydrodynamic flow [6]. As a result, integration of dialysis membrane is considered into the design of our new chip-based cartridge. Up to now, many approaches have been described to integrate membranes into glass/quartz or polymeric microfluidic chips. A simple method is direct incorporation by gluing or clamping commercial flat membranes. A major problem of this method is sealing, otherwise, a phenomenon of leakage around the membranes is always occurred due to the capillary force. A novel approach of sandwiching dialysis membrane was developed as schematically indicated in Figure 1. After optimizing IEF conditions, the separation of pI markers was performed on the obtained PDMS microfluidic chip. As exhibited in Figure 2a, six pI markers could be well separated on the PDMS chips patterned the channel of 100 μm deep, 100 μm wide by IEF-WCID. All the peaks were sharp and symmetric, indicating that both EOF and analytes adsorption were completely suppressed by the dynamic coating of PVP. The plots of peak position versus pI of these pI markers suggested good linearity of pH gradient (as shown in Figure 2b). The linear correlation coefficient was 0.9995 (n = 6). As expected to the capillary-based cartridge, the PDMS microfluidic chips could be applied for qualitative and quantitative analysis of proteins. Figure 3a exhibited that human hemoglobin control AFSC contains four known isoforms (HbA, HbF, HbS and HbC) mixed with two pI marker 6.14 and 8.18 were well separated on the PDMS chip by IEF-WCID, indicating the strong separation ability of chip similar to the commercial capillary-based cartridge. According to the linearity of pH gradient, these four isoforms with the pIs of 7.0, 7.1, 7.3 and 7.5, respectively, could be detected. An unknown isoform in human hemoglobin control marked asterisk in Figure 3A observed besides the definite four isoforms A, F, S and C. The myoglobin from horse heart contains two isoforms, whose pIs are 6.8 and 7.2, respectively. It can be seen from Figure 3b that these two isoforms were separated on PDMS chip by IEF-WCID. The peak 1 and 2 could be assigned to the two isoforms according to their pI. The pI of unknown peak marked asterisk could be measured to 6.25.