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Comparative Analysis of Static and Non-Static Assays for Biochemical Sensing Using On-Chip Integrated Field Effect Transistors and Solidly Mounted Resonators

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Comparative Analysis of Static and Non-Static Assays for Biochemical Sensing Using On-Chip Integrated Field Effect Transistors and Solidly Mounted Resonators

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Abstract

The advancement in micro/nanotechnologies has been continuously providing possibilities for inventing novel biochemical sensors. However, variations in the transducer type can cause different sensing results due to the differences in their mechanisms of analyzing biomolecular interactions. In this work, we focused on the comparative analysis of static and non-static assays for molecular interactions using on-chip integrated extended-gate field effect transistor (EGFET) as a static sensing interface and solidly mounted resonator (SMR) as a non-static sensing interface. Analysis of polyelectrolytes (PETs) surface assembly and antigen-antibody interaction using the two types of biochemical sensors presented consistent and complementary sets of information. Meanwhile, due to the difference in their operating mechanisms, variations on the detection efficiency, kinetics and thermodynamics were observed. Our results highlighted the critical dependence of signal detection on biochemical sensors’ operating mechanisms and provided a valuable guidance for static and non-static assays for biomolecular detections.

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... The integrated device was fabricated according to a previously published process (see ESI, † Fig. S1). 43 Before the fabrication of the resonator, a Bragg reflector was mounted on the silicon wafer by alternately depositing three pairs of silicon dioxide (SiO 2 ) and molybdenum (Mo) thin films. 44 The first layer of the Bragg reflector was used as the bottom electrode followed by the sputtering of highly c-axis oriented aluminum nitride (AlN) film as a piezoelectric layer and another layer of Mo as the top electrode. ...
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We developed an integrated device comprising a quartz crystal microbalance (QCM) and a field-effect transistor (FET) with a single common gold electrode in a flow chamber. An alternating current inducing oscillations in the piezoelectric quartz of the QCM sensor is electrically independent of the circuit for the FET output so that the two sensors in different detection mechanisms simultaneously record binding kinetics from a single protein solution on the same electrode. A conjunction of adsorbed mass from QCM with electric nature of bound protein from FET provided deeper understanding on a complex process of nonspecific protein adsorption and subsequent conformational changes at a solid/liquid interface. Lower apparent k(on) values obtained by FET than those obtained by QCM on hydrophobic surfaces are interpreted as preferred binding of protein molecules facing uncharged domains to the electrode surface, whereas higher k(off) values by FET than those by QCM imply active macromolecular rearrangements on the surfaces mainly driven by hydrophobic association in an aqueous medium. The advanced features of the combined sensor including in situ, label-free, and real-time monitoring provide information on structural dynamics, beyond measurements of affinities and kinetics in biological binding reactions.
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Very recently, electric-field-induced superconductivity in an insulator was realized by tuning charge carrier to a high density level (1 × 1014 cm−2). To increase the maximum attainable carrier density for electrostatic tuning of electronic states in semiconductor field-effect transistors is a hot issue but a big challenge. Here, ultrahigh density carrier accumulation is reported, in particular at low temperature, in a ZnO field-effect transistor gated by electric double layers of ionic liquid (IL). This transistor, called an electric double layer transistor (EDLT), is found to exhibit very high transconductance and an ultrahigh carrier density in a fast, reversible, and reproducible manner. The room temperature capacitance of EDLTs is found to be as large as 34 µF cm−2, deduced from Hall-effect measurements, and is mainly responsible for the carrier density modulation in a very wide range. Importantly, the IL dielectric, with a supercooling property, is found to have charge-accumulation capability even at low temperatures, reaching an ultrahigh carrier density of 8×1014 cm−2 at 220 K and maintaining a density of 5.5×1014 cm−2 at 1.8 K. This high carrier density of EDLTs is of great importance not only in practical device applications but also in fundamental research; for example, in the search for novel electronic phenomena, such as superconductivity, in oxide systems.
Article
Cellular production of such cytokines as interferon (IFN)-γ and tumor necrosis factor (TNF)-α is used to determine disease-specific immune responses and may be used to diagnose infectious diseases such as tuberculosis. In this paper, we describe the development of micropatterned electrodes functionalized with electroactive aptamers for multiplexed detection of immune-cell-produced cytokines. A sequence of electrode deprotection and aptamer incubation steps were used to assemble anti-IFN-γ DNA aptamers and anti-TNF-α RNA aptamers on individually addressable half-ring electrodes. Aptamer molecules were thiolated for assembly on gold and were functionalized with methylene blue redox reporter for electrochemical signal transduction. Specificity of individual sensors to the correct cytokine species was confirmed by exposure to recombinant cytokines. For cell detection experiments, electrode arrays were integrated into microfluidic devices and incubated with immune cells. Design of the surface was such that a small group of ~400 cells attached in the circular adhesion sites surrounded by half-ring electrodes sensing IFN-γ and TNF-α. The microdevice consisted of two parallel microfluidic channels, each channel containing four cell capture/sensing sites. Upon mitogenic activation, secreted IFN-γ and TNF-α molecules were monitored by performing square wave voltammetry (SWV) at different time points at individually addressable electrodes. This biosensing platform was used to analyze the quantity and rate of cytokine release from primary T cells and a monocyte cell line. Upon further development of this platform may be enhanced to enable detection of larger number of cytokines and used to correlate the levels and dynamics of cytokine release in immune cells to diagnosis and treatment of infectious diseases.
Article
Unlike the quartz crystal microbalance, which has been used extensively for the analysis of biochemical interactions, only few measurements with biochemical adsorbent have been done with film bulk acoustic resonators (FBAR). In this paper, the FBAR behaviour on exposure to a lipid vesicle solution and the formation of a polyelectrolyte multilayer structure is investigated and compared with the results obtained with the quartz crystal microbalance. Differences in the resonator response were found between the two techniques and depending on the resonators resonance frequency ranging from the MHz to the GHz regime. As an explanation, we suggest that the penetration depth and the influence on viscoelastic properties, which are both known to be frequency dependent, cause the variations in the results. As a consequence, the higher operating resonance frequencies of the FBAR increase the sensitivity to changes in the viscoelasticity of the adsorbent and also decrease the sensing length of the device.
Article
Alkanethiols are commonly used self-assembly reagents in the preparation of modified electrode. In this study, self-assembled monolayers (SAMs) of alkanethiols with different chain lengths formed on gold electrode were mainly investigated by electrochemical impedance spectroscopy (EIS) in addition to cyclic voltammetry (CV). Two types of immobilization strategies applied to the electrode were also examined. The interfacial properties of all self-assembled electrodes were evaluated in the presence of Fe(CN)63−/4− redox probe. A simple equivalent circuit model with a constant phase element was used to interpret the obtained impedance spectra. The results of cyclic voltammetry revealed that the voltammetric behavior of the redox probe was influenced by the electrode surface modification. The EIS data indicated that the formation of saturated SAMs on gold required at least 30 min and more apparent differences occurred with immobilization time compared to that obtained by CV. Depending on the alkyl chain length of the SAMs, the electron-transfer resistance increased and the capacitance decreased. It was found a good linear relationship between inverse capacitance and chain length for carboxyl-terminated SAMs. To achieve a defect-free structure and/or close packing, the direct application method may provide a more effective, rapid, and facile self-assembly immobilization than immersion method from the viewpoint of the electron-transfer resistance.
Article
This paper presents a review of acoustic-wave based MEMS devices that offer a promising technology platform for the development of sensitive, portable, real-time biosensors. MEMS fabrication of acoustic wave based biosensors enables device miniaturization, power consumption reduction and integration with electronic circuits. For biological applications, the biosensors are integrated in a microfluidic system and the sensing area is coated with a biospecific layer. When a bioanalyte interacts with the sensing layer, mass and viscosity variations of the biospecific layer can be detected by monitoring changes in the acoustic wave properties such as velocity, attenuation, resonant frequency and delay time. Few types of acoustic wave devices could be integrated in microfluidic systems without significant degradation of the quality factor. The acoustic wave based MEMS devices reported in the literature as biosensors and presented in this review are film bulk acoustic wave resonators (FBAR), surface acoustic waves (SAW) resonators and SAW delay lines. Different approaches to the realization of FBARs, SAW resonators and SAW delay lines for various biochemical applications are presented. Methods of integration of the acoustic wave MEMS devices in the microfluidic systems and functionalization strategies will be also discussed.
Article
Piezoelectric microelectromechanical systems (MEMS) resonant sensors, known for their excellent mass resolution, have been studied for many applications, including DNA hybridization, protein-ligand interactions, and immunosensor development. They have also been explored for detecting antigens, organic gas, toxic ions, and explosives. Most piezoelectric MEMS resonant sensors are acoustic sensors (with specific coating layers) that enable selective and label-free detection of biological events in real time. These label-free technologies have recently garnered significant attention for their sensitive and quantitative multi-parameter analysis of biological systems. Since piezoelectric MEMS resonant sensors do more than transform analyte mass or thickness into an electrical signal (e.g., frequency and impedance), special attention must be paid to their potential beyond microweighing, such as measuring elastic and viscous properties, and several types of sensors currently under development operate at different resonant modes (i.e., thickness extensional mode, thickness shear mode, lateral extensional mode, flexural mode, etc.). In this review, we provide an overview of recent developments in micromachined resonant sensors and activities relating to biochemical interfaces for acoustic sensors.
Article
Silicon nanowire field effect transistors (FETs) have emerged as ultrasensitive, label-free biodetectors that operate by sensing bound surface charge. However, the ionic strength of the environment (i.e., the Debye length of the solution) dictates the effective magnitude of the surface charge. Here, we show that control of the Debye length determines the spatial extent of sensed bound surface charge on the sensor. We apply this technique to different methods of antibody immobilization, demonstrating different effective distances of induced charge from the sensor surface.
Article
In this study, we have successfully demonstrated that a GaN nanowire (GaNNW) based extended-gate field-effect-transistor (EGFET) biosensor is capable of specific DNA sequence identification under label-free in situ conditions. Our approach shows excellent integration of the wide bandgap semiconducting nature of GaN, surface-sensitivity of the NW-structure, and high transducing performance of the EGFET-design. The simple sensor-architecture, by direct assembly of as-synthesized GaNNWs with a commercial FET device, can achieve an ultrahigh detection limit below attomolar level concentrations: about 3 orders of magnitude higher in resolution than that of other FET-based DNA-sensors. Comparative in situ studies on mismatches ("hotspot" mutations related to human p53 tumor-suppressor gene) and complementary targets reveal excellent selectivity and specificity of the sensor, even in the presence of noncomplementary DNA strands, suggesting the potential pragmatic application in complex clinical samples. In comparison with GaN thin film, NW-based EGFET exhibits excellent performance with about 2 orders higher sensitivity, over a wide detection range, 10(-19)-10(-6) M, reaching about a 6-orders lower detection limit. Investigations illustrate the unique and distinguished feature of nanomaterials. Detailed studies indicate a positive effect of energy band alignment at the biomaterials-semiconductor hybrid interface influencing the effective capacitance and carrier-mobility of the system.
Article
In this study, a highly sensitive and label-free multianalyte capacitive immunosensor was developed based on gold interdigitated electrodes (GID) capacitor arrays to detect a panel of disease biomarkers. C-reactive protein (CRP), TNFalpha, and IL6 have strong and consistent relationships between markers of inflammation and future cardiovascular risk (CVR) events. Early detection of a panel of biomarkers for a disease could enable accurate prediction of a disease risk. The detection of protein biomarkers was based on relative change in capacitive/dielectric properties. Two different lab-on-a-chip formats were employed for multiple biomarker detection on GID-capacitors. In format I, capacitor arrays were immobilized with pure forms of anti-CRP, -TNFalpha, and -IL6 antibodies in which each capacitor array contained a different immobilized antibody. Here, the CRP and IL6 were detected in the range 25 pg/ml to 25 ng/ml and 25 pg/ml to 1 ng/ml for TNFalpha in format I. Sensitive detection was achieved with chips co-immobilized (diluted) with equimolar mixtures of anti-CRP, -IL6, and -TNFalpha antibodies (format II) in which all capacitors in an array were identical and tested for biomarkers with sequential incubation. The resulting response to CRP, IL6, and TNFalpha in format II for all biomarkers was found to be within 25 pg/ml to 25 ng/ml range. The capacitive biosensor for panels of inflammation and CVR markers show significant clinical value and provide great potential for detection of biomarker panel in suspected subjects for early diagnosis.
Article
SPR imaging (SPRi) is at the forefront of optical label-free and real-time detection. It offers the possibility of monitoring hundreds of biological interactions simultaneously and from the binding profiles, allows the estimation of the kinetic parameters of the interactions between the immobilised probes and the ligands in solution. We review the current state of development of SPRi technology and its application including commercially available SPRi instruments. Attention is also given to surface chemistries for biochip functionalisation and suitable approaches to improve sensitivity.
Article
Electrochemical processes at the electrode-electrolyte (body fluid) interface are of ultimate importance for stimulating/sensing electrode function. A high electrode surface area is desirable for safe stimulation through double-layer charging and discharging. Pt and Pt-Ir alloys have been the most common electrode materials. The use of TiN coating as the surface layer on the electrode has found increasing interest because of its metal-like conductivity, excellent mechanical and chemical properties, and the fact that it can be deposited with a high surface area. In this work, electrochemical impedance spectroscopy (EIS), which is a sensitive and non-destructive technique and widely used for characterization of electrical properties of electrode-electrolyte interfaces, was applied to investigate pure Pt and Ti, and TiN coated electrodes exposed to a phosphate-buffered-saline (PBS) solution. Platinized Pt and Ti were also studied for comparison. The capacitance value of the electrodes in PBS was obtained through quantitative analysis of the EIS spectra. The results reveal that the capacitance of the TiN coated electrodes with a rough surface is several hundreds times higher than that of a smooth Pt surface. Platinization of Ti can also increase the capacitance to the same extent as platina. EIS has been shown to be a powerful technique for characterization of stimulating/sensing electrodes.
Article
The labelfree detection of nucleic acid sequences is one of the modern attempts to develop quick, cheap and miniaturised hand-held devices for the future genetic testing in biotechnology and medical diagnostics. We present an approach to detect the hybridisation of DNA sequences using electrolyte-oxide-semiconductor field-effect transistors (EOSFETs) with micrometer dimensions. These semiconductor devices are sensitive to electrical charge variations that occur at the surface/electrolyte interface, i.e. upon hybridisation of oligonucleotides with complementary single-stranded (ss) oligonucleotides, which are immobilised on the oxide surface of the transistor gate. This method allows direct, time-resolved and in situ detection of specific nucleic acid binding events without any labelling. We focus on the detection mechanism of our sensors by using oppositely charged polyelectrolytes (PAH and PSS) subsequently attached to the transistor structures. Our results indicate that the sensor output is charge sensitive and distance dependent from the gate surface, which pinpoints the need for very defined surface chemistry at the device surface. The hybridisation of natural 19 base-pair sequences has been successfully detected with the sensors. In combination with nano-transistors a PCR free detection system might be feasible in future.
Article
Electrostatic layer-by-layer film assembly is an attractive way to non-covalently incorporate proteins and bioactive moieties into the surface of conventional biomaterials. Selection of polycationic and polyanionic components and deposition conditions can be used to control the interfacial properties, and through them protein adsorption, cell adhesion, and tissue development. In this study the polycation was poly(allylamine hydrochloride) (PAH), which is a weak base and consequently adsorbs at interfaces in a pH-dependent manner, and the polyanion was heparin, which is capable of interacting with many adhesion ligands and growth factors. PAH/heparin multilayer films were formed using PAH solutions of pH 6.4, 7.4, 8.4, and 9.4. Film thickness increased both with the number of PAH/heparin bilayers and the pH of the PAH solution. Films consisting of 10 bilayers with heparin topmost exhibited similar bulk atomic compositions and penetration of PAH into the heparin top layer. Finally, fibronectin adsorption and cell adhesion were maximal at an intermediate pH (pH 8.4>pH 9.4>pH 7.4). These results demonstrate that heparin-containing electrostatic films support cell adhesion and protein adsorption in a manner sensitive to film deposition conditions.
Article
The dispersion of nanotubes by pH-responsive polymers (i.e., weak polyelectrolytes) enables the macroscopic properties of aqueous suspensions to be tuned. Microstructural changes were achieved as a function of pH in aqueous suspensions containing single-walled carbon nanotubes and imaged by cryogenic-TEM. Clear evidence of pH-sensitive nanotube dispersion is shown. We expect that many useful properties of these nanotube-polymer systems could be sensitive to microstructure, making this technique important for aqueous processing of carbon nanotubes and macroscopic tailoring of solid polymer nanocomposite behavior.
Article
Nanowire field effect transistors (NW-FETs) can serve as ultrasensitive detectors for label-free reagents. The NW-FET sensing mechanism assumes a controlled modification in the local channel electric field created by the binding of charged molecules to the nanowire surface. Careful control of the solution Debye length is critical for unambiguous selective detection of macromolecules. Here we show the appropriate conditions under which the selective binding of macromolecules is accurately sensed with NW-FET sensors.
  • L Mu
  • I A Droujinine
  • N K Rajan
  • S D Sawtelle
  • M A Reed
L. Mu, I.A. Droujinine, N.K. Rajan, S.D. Sawtelle, M.A. Reed, Direct, Rapid, and Label-Free Detection of Enzyme–Substrate Interactions in Physiological Buffers Using CMOS-Compatible Nanoribbon Sensors, Nano letters, 14(2014) 5315-22.
Buildup of ultrathin multilayer films by a self-assembly process
  • G Decher
  • J Hong
  • J Schmitt
G. Decher, J. Hong, J. Schmitt, Buildup of ultrathin multilayer films by a self-assembly process: