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ABSTRACT: Aim: To demonstrate a label-free electrical immunoassay for profiling vascular biomarker N-terminal pro-brain natriuretic peptide (NT-proBNP) associated with improved cardiac risk prediction. Materials & methods: A high-density nanowell-based electrical immunoassay has been designed by integrating nanoporous aluminum oxide onto printed circuit board chips for the detection of NT-proBNP. The concentration of the biomarker is quantitatively determined by measuring impedance changes to the electrical double layer within the nanowells using electrochemical impedance spectroscopy. Detection sensitivity in the fg/ml range was obtained due to spatial confinement of the target biomarkers in size-matched nanowells. Results & discussion: Electrical immunoassay performance was determined for the detection of NT-proBNP in phosphate-buffered saline (PBS) and human serum (HS). The lower limit of detection for the sensor was observed to be 10 fg/ml in PBS and 500 fg/ml in HS. The upper limit of detection was observed to be 500 fg/ml in PBS and 500 ng/ml in HS. Conclusion: A label-free technique for detection of NT-proBNP at clinically relevant concentrations for evaluating cardiac risk is demonstrated. High sensitivity and specificity, robust detection and low volume (100 µl) per assay project the technology to be a successful competitor to traditional ELISA-based techniques.
Future Cardiology 01/2013; 9(1):137-47.
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Advanced Science, Engineering and Medicine. 12/2012;
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ABSTRACT: A silicon nanosensor technology based on electrical impedance measurements has been developed for the detection of proteins. The nanosensor miniaturizes the high-density, low-volume multiwell plate concept. The scientific core of this technology lies in integrating nanoporous membranes with microfabricated chip platforms. This results in the conversion of individual pores into nanowells of picoliter volume. Monoclonal antibodies were localized and isolated into individual wells. Detection of two cardiac proteomic biomarkers has been demonstrated with this technology. The two proteins, C-reactive protein and NT-pro-brain natriuretic peptide (BNP), are associated with adverse cardiac outcomes in clinical samples when detected in the pg/mL concentration. The formation of the antibody-antigen binding complex occurs in individual wells. The membrane allows for robust separation among individual wells. This technology has the capability to achieve near real-time detection with improved sensitivity at 1 ag/mL for BNP and 1 fg/mL for CRP from human serum.
Journal of the Association for Laboratory Automation 09/2012; · 1.42 Impact Factor
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ABSTRACT: an electrochemical approach towards identifying antigen - antibody binding interactions is studied by using a non-faradaic impedimetric sensor fabricated on a printed circuit board (PCB) chip. An electrical methodology for detecting protein interactions at ultra-low concentrations (in the femtogram/mL) regime has been demonstrated. Nanoporous alumina with pore diameter of 200nm and pore depth of 250 nm was used as the signal amplifying medium. Cardiac biomarker, brain natriuretic peptide (BNP) was used as the study marker in characterizing the sensor's sensitivity. A sensitivity of 10 femtogram/mL was determined based on the impedimetric signal response. Sensitivity was determined through Nyquist plot analysis for the non-faradaic interactions of the protein biomolecules. This paper is the first demonstration of clinically relevant limit of detection with the BNP biomarker.
Conference proceedings: ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Conference 08/2012; 2012:3251-4.
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ABSTRACT: Trace contamination of ground water sources has been a problem ever since the introduction of high-soil-mobility pesticides, one such example is atrazine. In this paper we present a novel nanoporous portable bio-sensing device that can identify trace contamination of atrazine through a label-free assay. We have designed a pesticide sensor comprising of a nanoporous alumina membrane integrated with printed circuit board platform. Nanoporous alumina in the biosensor device generates a high density array of nanoscale confined spaces. By leveraging the size based immobilization of atrazine small molecules we have designed electrochemical impedance spectroscopy based biosensor to detect trace amounts of atrazine. We have calibrated the sensor using phosphate buffered saline and demonstrated trace detection from river and bottled drinking water samples. The limit of detection in all the three cases was in the femtogram/mL (fg/mL) (parts-per-trillion) regime with a dynamic range of detection spanning from 10 fg/mL to 1 ng/mL (0.01 ppt to 1 ppm). The selectivity of the device was tested using a competing pesticide; malathion and selectivity in detection was observed in the fg/mL regime in all the three cases.
Biosensors & bioelectronics 02/2012; 32(1):155-62. · 5.43 Impact Factor
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ABSTRACT: Protein misfolding and aggregation is a critically important feature in many devastating neurodegenerative diseases, therefore characterization of the CSF concentration profiles of selected key forms and morphologies of proteins involved in these diseases, including β-amyloid (Aβ) and α-synuclein (a-syn), can be an effective diagnostic assay for these diseases. CSF levels of tau and Aβ have been shown to have great promise as biomarkers for Alzheimer's disease. However since the onset and progression of many neurodegenerative diseases have been strongly correlated with the presence of soluble oligomeric aggregates of proteins including various Aβ and a-syn aggregate species, specific detection and quantification of levels of each of these different toxic protein species in CSF may provide a simple and accurate means to presymptomatically diagnose and distinguish between these diseases. Here we show that the presence of different protein morphologies in human CSF samples can be readily detected using highly selective morphology specific reagents in conjunction with a sensitive electronic biosensor. We further show that these morphology specific reagents can readily distinguish between post-mortem CSF samples from AD, PD and cognitively normal sources. These studies suggest that detection of specific oligomeric aggregate species holds great promise as sensitive biomarkers for neurodegenerative disease.
Integrative Biology 11/2011; 3(12):1188-96. · 4.51 Impact Factor
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IEEE Nano 2011; 08/2011
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IEEE Nano 2011; 08/2011
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ABSTRACT: “Label-free” biomolecule sensors for detection of inflammatory cardiovascular biomarker associated with vulnerable coronary vascular plaque were designed and fabricated using micro and nano-textured polystyrene structures that functioned as sensing elements coupled with electronic measurement equipment. We demonstrated that scaling down the surface texturing from the micro to the nanoscale enhances the amplitude of the measured signal strength. We believe that the nanoscale fiber morphology provides size matched spaces for trapping and immobilizing the protein biomolecules resulting in enhanced detection and signal strength. We selected polystyrene as the model system and demonstrated the detection of human serum C-reactive protein (hs-CRP). We employed these findings in designing a platform “lab-on-a-chip” protein sensor. Comparative studies were performed on two different polystyrene textured surfaces: a polystyrene microsphere mat, and an electrospun polystyrene nanofiber matrix.
MRS Proceedings. 12/2010; 1358.
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08/2010: pages 47 - 90; , ISBN: 9780470622551
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ABSTRACT: The goal of our research is to develop an ultrasensitive diagnostic platform called 'NanoMonitor' to enable rapid label-free analysis of a highly promising class of biomarkers called glycans (oligosaccharide chains attached to proteins) with high sensitivity and selectivity. The glycosylation of fetuin - a serum protein - and extracts from a human pancreatic cancer line was analyzed to demonstrate the capabilities of the NanoMonitor.
The NanoMonitor device consists of a silicon chip with an array of gold electrodes forming multiple sensor sites and works on the principle of electrochemical impedance spectroscopy. Each sensor site is overlaid with a nanoporous alumina membrane that forms a high density of nanowells on top of each electrode. Lectins (proteins that bind to and recognize specific glycan structures) are conjugated to the surface of the electrode. When specific glycans from a test sample bind to lectins at the base of each nanowell, a perturbation of electrical double-layer occurs, which results in a change in the impedance. Using the lectins Sambucs nigra agglutinin (SNA) and Maackia amurensis agglutinin (MAA), subtle variations to the glycan chains of fetuin were investigated. Protein extracts from BXPC-3, a cultured human pancreatic cancer cell line were also analyzed for binding to SNA and MAA lectins. The performance of the NanoMonitor was compared to a conventional laboratory technique: lectin-based enzyme linked immunosorbent assay (ELISA).
The NanoMonitor was used to identify glycoform variants of fetuin and global differences in glycosylation of protein extracts from cultured human pancreatic cancerous versus normal cells. While results from NanoMonitor correlate very well with results from lectin-based ELISA, the NanoMonitor is rapid, completely label free, requires just 10 microl of sample, is approximately five orders of magnitude more sensitive and highly selective over a broad dynamic range of glycoprotein concentrations.
Based on its performance metrics, the NanoMonitor has excellent potential for development as a point-of-care handheld electronic biosensor device for routine detection of glycan biomarkers from clinical samples.
Nanomedicine 04/2010; 5(3):369-78. · 5.05 Impact Factor
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ABSTRACT: The goal of our research is to demonstrate the feasibility of employing biogenic nanoporous silica as a key component in developing a biosensor platform for rapid label-free electrochemical detection of cardiovascular biomarkers from pure and commercial human serum samples with high sensitivity and selectivity. The biosensor platform consists of a silicon chip with an array of gold electrodes forming multiple sensor sites and works on the principle of electrochemical impedance spectroscopy. Each sensor site is overlaid with a biogenic nanoporous silica membrane that forms a high density of nanowells on top of each electrode. When specific protein biomarkers: C-reactive protein (CRP) and myeloperoxidase (MPO) from a test sample bind to antibodies conjugated to the surface of the gold surface at the base of each nanowell, a perturbation of electrical double layer occurs resulting in a change in the impedance. The performance of the biogenic silica membrane biosensor was tested in comparison with nanoporous alumina membrane-based biosensor and plain metallic thin film biosensor. Significant enhancement in the sensitivity and selectivity was achieved with the biogenic silica biosensor, in comparison to the other two, for detecting the two protein biomarkers from both pure and commercial human serum samples. The sensitivity of the biogenic silica biosensor is approximately 1 pg/ml and the linear dose response is observed over a large dynamic range from 1 pg/ml to 1 microg/ml. Based on its performance metrics, the biogenic silica biosensor has excellent potential for development as a point of care handheld electronic biosensor device for detection of protein biomarkers from clinical samples.
Biosensors & bioelectronics 04/2010; 25(10):2336-42. · 5.43 Impact Factor
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ABSTRACT: "Label-free" biomolecule sensors for detection of inflammatory cardiovascular biomarker associated with vulnerable coronary vascular plaque rupture were designed and fabricated using micro- and nanotextured polystyrene (PS) polymer structures that functioned as sensing elements coupled with electronic measurement equipment. We demonstrated that scaling down the surface texturing from the micro- to the nanoscale enhances the amplitude of the measured detected signal strength. We believe that the nanoscale fiber morphology provides size-matched spaces for trapping and immobilizing the protein biomolecule, resulting in improved detection signal strength. We selected PS as the model system and demonstrated the detection of human serum C-reactive protein. We employed these findings in designing a platform "lab-on-a-chip" protein sensor. Comparative studies were performed on PS textured surfaces of two different surface features: a PS microsphere mat and an electrospun PS nanofiber matrix. FROM THE CLINICAL EDITOR: In this study, nanotechnology-based biosensors for vulnerable coronary vascular plaque rupture were designed and fabricated using micro- and nanotextured polystyrene polymer structures. The authors demonstrated that scaling down the surface texturing from the micro- to the nanoscale enhances the sensitivity of this detection method.
Nanomedicine: nanotechnology, biology, and medicine 03/2010; 6(5):642-50. · 5.44 Impact Factor
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ABSTRACT: In this paper we present the design, fabrication and development of an optical, label free chemical sensor technology based on the portable "lab-on-a-chip" format. This sensor technology employs the use of nanotextured thin film surfaces packaged in a stacked vertical array format functioning as organic light emitting diodes (OLED's). The intensity of emitted light from OLED's is modulated as a function of the analyte concentration. The OLEDs were fabricated in a layer by layer configuration with an indium tin oxide anode and an aluminum cathode and TPD as a hole transport layer and AIQ3 as an electron transport layer. ITO/TPD/AlQ3/Al sandwich OLEDs were converted into sensors by converting the cathode (Al) surface into an active sensing area. The prototype sensor performance was evaluated in the detection of two aliphatic hydrocarbons-ethanol and methanol. The detection sensitivity was found to be in the lower parts per million (ppm). The limit of detection for ethanol was 1 ppm and that for methanol was 10 ppm. Chemical detection was achieved upon the comparison of turn-on voltages and the intensities of the output light from the OLED when chemical was being injected onto the cathode surface, with that of a standard OLED turn-on voltage and intensity. The modulation in these two parameters with respect to the standard was determined as a measure of detection of the two chemical species.
Journal of Nanoscience and Nanotechnology 11/2009; 9(11):6299-306. · 1.56 Impact Factor
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ABSTRACT: The objective of this research is to demonstrate the potential of iridium oxide (IrOx) nanowires based device towards detection of proteins that are disease biomarkers. This device is based on electrical detection of protein biomarkers wherein an immunoassay is built onto the iridium oxide nanowires that in turn undergoes specific electrical parameter perturbations during each binding event associated with the immunoassay. Detection of two inflammatory proteins C-reactive protein (CRP) and Myeloperoxidase (MPO) that are biomarkers of cardiovascular diseases is demonstrated. The performance metrics of the device in response to the two biomarkers in pure form and in serum samples were evaluated and compared to standard ELISA assays. The methodology that has been adopted is based on measuring impedance and calibrating its change in magnitude with concentration of proteins. We demonstrate the following performance metrics: limits of detection up to 1 ng/ml for CRP and 500 pg/ml for MPO in pure and serum samples; linear dynamic range of detection from 10 ng/ml to 100 microg/ml for CRP and 1 ng/ml to 1 microg/ml for MPO and cross-reactivity contained at less than 10% of selective binding for both the inflammatory proteins. Iridium oxide has an ability to detect very small changes to the surface charge and this capability is utilized for achieving the performance metrics and forms the basis of the key innovations of this technology, which are, improving the selectivity and sensitivity of detection.
Biosensors & bioelectronics 05/2009; 24(10):3078-83. · 5.43 Impact Factor
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12/2008: pages 345-375;
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ABSTRACT: The method for patterning arrays of multiwalled carbon nanotubes (MWCNT's) in symmetric patterns to form junctions has been demonstrated. This has been achieved by incorporating the technique of microcontact printing using poly-dimethylsiloxane (PDMS) molds. Relief structures in the order of a few micrometers were fabricated that enabled the transfer of continuous horizontal arrays of MWCNT's in aqueous suspension in a controlled manner. The MWCNT's were patterned onto silicon microelectrode substrates with metallic gold electrodes. These were fabricated using standard photolithography techniques. The silicon substrates served as a base platform with suitable measurement microelectrodes for electrically characterizing the crossbar junction arrays. Using a dual alignment and stamping process, PDMS molds were inked alternatively with p-type and n-type suspensions of MWCNT's and transferred in a grid-like manner onto the base platform. Parallel alignment of the MWCNT's was achieved due to the geometry of the mold relief structures. This step-by-step assembly resulted in the formation of crossbar MWCNT array structures. Each of these crosspoints in the individual junction can function as an addressable crossbar nanodevice. The functionality of this circuit was demonstrated through the current-voltage (I-V) characteristics. Using these high-density crossarray circuit patterns, addressable nanostructures that form the building blocks of highly integrated device arrays can be built.
Journal of Nanoscience and Nanotechnology 05/2008; 8(4):1951-8. · 1.56 Impact Factor
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ABSTRACT: A novel electronic spectroscopy technique based on dipole-dipole interactions for the identification of chemical analytes has been developed. This technique is based on the measurement of the charge transfer of chemical analytes to a multiwalled carbon nanotube mat-based sensing system. This technique was used for the identification of three aromatic hydrocarbons, namely, benzene, toluene, and xylene, at 100 parts-per-billion concentration. This technique was evaluated with multiwalled carbon nanotube mats for rapid, reliable, and robust identification of the three chemicals that belong to the same genre. The technique involves the identification of electronic spectral signatures of these chemicals using frequency domain analysis of the voltage signals generated by the binding of the chemical analytes onto the multiwalled carbon nanotube mat surfaces. This technique has the potential for rapid and accurate identification of multiple chemical analytes in a multiplexed fashion using a single-sensor device. In addition, this particular device configuration in conjunction with the electronic dipole spectroscopy results is a powerful lab-on-a-chip device for chemical and biological sensing applications.
Chemical Engineering Communications 02/2008; 195(2):115-128. · 0.95 Impact Factor
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ABSTRACT: The immobilization of biomolecules on a solid substrate and their localization in “small” regions are major requirements for a variety of biomedical diagnostic applications, where rapid and accurate identification of multiple biomolecules is essential. In this specific application we have fabricated nanomitors for identifying specific protein biomarkers based on the electrical detection of antibody-antigen binding events.The nanomonitor, lab-on-a-chip device technology is based on electrical detection of protein biomarkers. It is based on developing high density, low volume multi-well plate devices. The scientific core of this technology lies in integrating nanomaterial with micro fabricated chip platforms and exploiting the improve surface area to volume to improve the detection.The devices that have been developed utilize electrical detection mechanisms where capacitance and conductance changes due to protein binding are used as “signatures” for biomarker profiling. In comparison to optical methods, the electrical detection technique is non-invasive as well as a label free. The signal acquisition is simple and it uses the existing data acquisition and signal analysis methods
MRS Proceedings. 12/2007; 1095.
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ABSTRACT: The paper presents an evaluation of the feasibility of developing ionic surfactant coated single walled carbon nanotube (SWCNTs) (P-N) junction clusters via microcontact printing using intrinsic semi conducting SWCNTs. These SWCNTs are doped with anionic and cationic surfactant molecules respectively, thereby altering the Fermi energy levels of and its electrical properties. Two types of surfactants were used for doping the SWCNTs to develop extrinsically doped P and N type SWCNTs. Sodium dodecyl sulfate (SDS) having Na+ positive ionic charge (anions) and Cetyl trimethylammonium bromide (CTAB), having Br- negative ionic charge (cations) on its hydrophilic ends have been used to generate anionic SWCNTs (P type) and cationic SWCNTs( N- type respectively.
Using, dual level patterning process, these extrinsically doped anionic and cationic semiconducting SWCNTs clusters are alternatively symmetrically patterned in a parallel array to form crossbar P-N junctions onto a standard microfabricated platform using flexible polymeric poly-dimethylsiloxane (PDMS) stamps. Ink-based transfer of the nanomaterial from the relief structures achieves parallel alignment of SWCNTs clusters. The electrical device characterization is achieved by measuring I-V characteristics from the base micro fabricated platform.
Functionality of the nanodevice is demonstrated by studying the rectifying current – voltage (I-V) characteristics that shows promise towards the formation of a junction diode array, which can be used for integrating complex logic devices for high-end applications such as memory module and addressable logic. Lastly, we believe that the chemical modulation method and microcontact printing techniques will have a wide scope for development of nanomaterial-based devices.
MRS Proceedings. 12/2007; 1081.
