ABTECH Scientific Inc.

Richmond, VA, United States

ABTECH Scientific Inc.

Richmond, VA, United States
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Kotanen C.N.,Clemson University | Moussy F.G.,Brunel University | Carrara S.,Ecole Polytechnique Federale de Lausanne | Guiseppi-Elie A.,Clemson University | Guiseppi-Elie A.,ABTECH Scientific Inc.
Biosensors and Bioelectronics | Year: 2012

The implantable enzyme amperometric biosensor continues as the dominant in vivo format for the detection, monitoring and reporting of biochemical analytes related to a wide range of pathologies. Widely used in animal studies, there is increasing emphasis on their use in diabetes care and management, the management of trauma-associated hemorrhage and in critical care monitoring by intensivists in the ICU. These frontier opportunities demand continuous indwelling performance for up to several years, well in excess of the currently approved seven days. This review outlines the many challenges to successful deployment of chronically implantable amperometric enzyme biosensors and emphasizes the emerging technological approaches in their continued development. The foreign body response plays a prominent role in implantable biotransducer failure. Topics considering the approaches to mitigate the inflammatory response, use of biomimetic chemistries, nanostructured topographies, drug eluting constructs, and tissue-to-device interface modulus matching are reviewed. Similarly, factors that influence biotransducer performance such as enzyme stability, substrate interference, mediator selection and calibration are reviewed. For the biosensor system, the opportunities and challenges of integration, guided by footprint requirements, the limitations of mixed signal electronics, and power requirements, has produced three systems approaches. The potential is great. However, integration along the multiple length scales needed to address fundamental issues and integration across the diverse disciplines needed to achieve success of these highly integrated systems, continues to be a challenge in the development and deployment of implantable amperometric enzyme biosensor systems. © 2012 Elsevier B.V.


Yang L.,North Carolina Central University | Guiseppi-Wilson A.,ABTECH Scientific Inc. | Guiseppi-Elie A.,ABTECH Scientific Inc. | Guiseppi-Elie A.,Clemson University
Biomedical Microdevices | Year: 2011

Microlithographically fabricated interdigitated microsensor electrodes (IMEs) were cleaned, surface activated, chemically functionalized (amine) and derivatized with an Acrloyl-PEG-NHS to receive a spun-applied monomer cocktail of UV polymerizable monomer. IMEs were 2050.5, 1550.5, 1050.5 and 0550.5 possessing lines and spaces that were 20, 15, 10, and 5 μm respectively; 5 mm line lengths and were 50 lines on each opposing bus. Bioactive hydrogels were synthesized from spun-applied and UV-crosslinked tetraethyleneglycol diacrylate (TEGDA) (crosslinker), 2-hydroxyethylmethacrylate (HEMA), polyethyleneglycol (200) monomethacrylate (PEGMA), N-[tris(hydroxymethyl) methyl]-acrylamide (HMMA) and poly(HEMA) (MW 60,000) (viscosity modifier) and 2,2-dimethoxy-2- phenylacetophenone (DMPA) (photoinitiator) to produce a 5 μm thick p(HEMA-co-PEGMA-co-HMMA) hydrogel membrane on the IMEs. Unmodified and hydrogel coated IMEs where characterized by AC electrical impedance spectroscopy using 50 mV p-t-p over the frequency range from 10 Hz to 100 kHz in aqueous PBS 7.4 buffer and in buffer containing 50 mM [Fe(CN)6]3-/4- solution at RT. Impedimetric responses were found to scale with the device geometric parameters. Equivalent circuit modeling revealed deviations from ideality at lower device dimensions suggesting an implication of the substrate surface charge on the double layer capacitance of the electrodes. Diffusion coefficients derived from the Warburg component are in accord with literature values. © Springer Science+Business Media, LLC 2010.


Kotanen C.N.,Clemson University | Wilson A.N.,Clemson University | Dong C.,West Virginia University | Dinu C.-Z.,West Virginia University | And 3 more authors.
Biomaterials | Year: 2013

The physicochemical properties of soft electrode materials for the abio-bio interface of advanced biosensors and next generation bionic devices in the form of electroconductive hydrogels (ECH) of interpenetrating networks of polypyrrole formed within poly(hydroxyethylmethacrylate)-based hydrogels were examined. The 1.5mol% UV-crosslinked tetraethyleneglycol diacrylate (TEGDA) (step 1) poly(HEMA) and the electropolymerized (step 2) polypyrrole co-networks were covalently joined by the inclusion of a bifunctional monomer (1.5mol%), 2-methacryloyloxyethyl-4(3-pyrrolyl)butanate (MPB) that served to covalently link the two networks. The optical absorbance, degree of hydration, the frequency dependent electrical impedance and the elastic modulus were examined as a function of electropolymerization charge density (step 2) (1-900mC/cm2) used to prepare the linked, interpenetrating co-networks. The absorption at 430nm showed a monotonic increase with electropolymerization charge density and correlated with the increase in elastic modulus [56 (±32)-499 (±293) kPa], the decrease in % hydration (68-0%) and the decrease in membrane electrical resistance. Polypyrrole (PPy) grows initially from the gel-electrode interface to fill voids within the hydrogel and ultimately onto the surface of the hydrogel. Growth of attachment dependent Rhabdomyosarcoma (RMS13) and pheochromocytoma (PC 12) cells reflects this evolution, showing an increase to a maximal value and then to decrease again at high electropolymerization charge density. © 2013 Elsevier Ltd.


Guiseppi-Elie A.,Clemson University | Guiseppi-Elie A.,ABTECH Scientific Inc.
Analytical and Bioanalytical Chemistry | Year: 2011

Following hemorrhage-causing injury, lactate levels rise and correlate with the severity of injury and are a surrogate of oxygen debt. Posttraumatic injury also includes hyperglycemia, with continuously elevated glucose levels leading to extensive tissue damage, septicemia, and multiple organ dysfunction syndrome. A temporary, implantable, integrated glucose and lactate biosensor and communications biochip for physiological status monitoring during hemorrhage and for intensive care unit stays has been developed. The dual responsive, amperometric biotransducer uses the microdisc electrode array format upon which were separately immobilized glucose oxidase and lactate oxidase within biorecognition layers, 1.0-5.0 μm thick, of 3 mol% tetraethyleneglycol diacrylate cross-linked p(HEMA-co-PEGMA-co-HMMA-co-SPA)-p(Py-co-PyBA) electroconductive hydrogels. The device was then coated with a bioactive hydrogel layer containing phosphoryl choline and polyethylene glycol pendant moieties [p(HEMA-co-PEGMA-co-HMMA-co-MPC)] for indwelling biocompatibility. In vitro cell proliferation and viability studies confirmed both polymers to be non-cytotoxic; however, PPy-based electroconductive hydrogels showed greater RMS 13 and PC12 proliferation compared to controls. The glucose and lactate biotransducers exhibited linear dynamic ranges of 0.10-13.0 mM glucose and 1.0-7.0 mM and response times (t 95) of 50 and 35-40 s, respectively. Operational stability gave 80% of the initial biosensor response after 5 days of continuous operation at 37 °C. Preliminary in vivo studies in a Sprague-Dawley hemorrhage model showed tissue lactate levels to rise more rapidly than systematic lactate. The potential for an implantable biochip that supports telemetric reporting of intramuscular lactate and glucose levels allows the refinement of resuscitation approaches for civilian and combat trauma victims. © 2010 Springer-Verlag.


Kotanen C.N.,Clemson University | Tlili C.,Clemson University | Guiseppi-Elie A.,Clemson University | Guiseppi-Elie A.,ABTECH Scientific Inc.
Talanta | Year: 2013

Fabrication of an enzyme amperometric biosensor for glucose via electropolymerization of pyrrole in the presence of glucose oxidase onto a hydrogel coated platinum electrode is hereby established as a viable biotransducer fabrication method. Platinum micro- (φ=25 mm) and macro- (φ=100 μm) electrodes were electrochemically activated and chemically modified with 3-aminopropyl-trimethoxysilane (APTMS), functionalized with acryloyl(polyethyleneglycol)-N-hydroxysuccinamide (ACRL-PEG-NHS), dipped into a polyHEMA based hydrogel cocktail and UV cross-linked. Electropolymerization of Py in the presence of GOx produced glucose responsive biotransducers that showed; (i) a 4-fold reduction in sensitivity compared with directly electropolymerized PPy films, (ii) an electropolymerization charge density dependence of biotransducer sensitivity and enzyme activity that was maximal at 1.0 mC/cm2 with an apparent KM of 33 mM, (iii) interference screening of ascorbic acid and (iv) a temporal increase in sensitivity with storage over a 17 days period. This method has the ability to precisely and quantitatively add enzyme catalytic bioactivity to metal or semiconductor biointerfaces for applications in biosensors, bioelectronics and bionics. © 2012 Elsevier B.V. All rights reserved.


Wilson A.N.,Clemson University | Guiseppi-Elie A.,Clemson University | Guiseppi-Elie A.,ABTECH Scientific Inc.
International Journal of Pharmaceutics | Year: 2014

A drug delivery platform comprising a biocompatible, bioresponsive hydrogel and possessing a covalently tethered peptide-drug conjugate was engineered to achieve stasis, via a closed control loop, of the external biochemical activity of the actuating protease. The delivery platform contains a peptide-drug conjugate covalently tethered to the hydrogel matrix, which in the presence of the appropriate protease, was cleaved and the drug released into the bathing environment. This platform was developed and investigated in silico using a finite element modeling (FEM) approach. Firstly, the primary governing phenomena guiding drug release profiles were investigated, and it was confirmed that under transport-limited conditions, the diffusion of the enzyme within the hydrogel and the coupled enzyme kinetics accurately model the system and are in agreement with published results. Secondly, the FEM model was used to investigate the release of a competitive protease inhibitor, MAG283, via cleavage of Acetyl-Pro-Leu-Gly|Leu-MAG-283 by MMP9 in order to achieve targeted homeostasis of MMP-9 activity, such as in the pathophysiology of chronic wounds, via closed-loop feedback control. The key engineering parameters for the delivery device are the radii of the hydrogel microspheres and the concentration of the peptide-inhibitor conjugate. Homeostatic drug delivery, where the focus turns away from the drug release rate and turns toward achieving targeted control of biochemical activity within a biochemical pathway, is an emerging approach in drug delivery methodologies for which the potential has not yet been fully realized. © 2013 Elsevier B.V. All rights reserved.


Wilson A.N.,Clemson University | Guiseppi-Elie A.,Clemson University | Guiseppi-Elie A.,ABTECH Scientific Inc.
Advanced Healthcare Materials | Year: 2013

Bioresponsive hydrogels are emerging with technological significance in targeted drug delivery, biosensors, and regenerative medicine. Their ability to respond to specific biologically derived stimuli creates a design challenge in effectively linking the conferred biospecificity with an engineered response tailored to the needs of a particular application. Moreover, the fundamental phenomena governing the response must support an appropriate dynamic range, limit of detection, and the potential for feedback control. The design of these systems is inherently complicated due to the high interdependency of the governing phenomena that guide sensing, transduction, and actuation of the hydrogel. Future advancements in bioresponsive hydrogels will out of necessity contain control loops similar to synthetic metabolic pathways. The use of these materials will continue to expand as they become coupled and integrated with new technologies. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Karunwi O.,Clemson University | Guiseppi-Elie A.,Clemson University | Guiseppi-Elie A.,ABTECH Scientific Inc.
Journal of Nanobiotechnology | Year: 2013

Background: Generation-3 (Gen-3) biosensors and advanced enzyme biofuel cells will benefit from direct electron transfer to oxidoreductases facilitated by single-walled carbon nanotubes (SWNTs).Methods: Supramolecular conjugates of SWNT-glucose oxidase (GOx-SWNT) were produced via ultrasonic processing. Using a Plackett-Burman experimental design to investigate the process of tip ultrasonication (23 kHz), conjugate formation was investigated as a function of ultrasonication times (0, 5, 60 min) and functionalized SWNTs of various tube lengths (SWNT-X-L), (X = -OH or -COOH and L = 3.0 μm, 7.5 μm).Results: Enzyme activity (KM, kcat, kcat/KM, vmax and n (the Hill parameter)) of pGOx (pristine), sGOx (sonicated) and GOx-SWNT-X-L revealed that sonication of any duration increased both KM and kcat of GOx but did not change kcat/KM. Functionalized tubes had the most dramatic effect, reducing both KM and kcat and reducing kcat/KM. UV-vis spectra over the range of 300 to 550 nm of native enzyme-bound FAD (λmax at 381 and 452 nm) or the blue-shifted solvated FAD of the denatured enzyme (λmax at 377 and 448 nm) revealed that ultrasonication up to 60 minutes had no influence on spectral characteristics of FAD but that the longer SWNTs caused some partial denaturation leading to egress of FAD. Circular dichroism spectral analysis of the 2° structure showed that sonication of any duration caused enrichment in the α-helical content at the sacrifice of the unordered sequences in GOx while the presence of SWNTs, regardless of length and/or functionality, reduced the β-sheet content of pristine GOx. Surface profiling by white light interferometry revealed that ultrasonication produced some aggregation of GOx and that GOx effectively debundled the SWNT.Conclusions: Supramolecular conjugates formed from shorter, -OH functionalized SWNTs using longer sonication times (60 min) gave the most favored combination for forming bioactive conjugates. © 2013 Karunwi and Guiseppi-Elie; licensee BioMed Central Ltd.


Guiseppi-Elie A.,ABTECH Scientific Inc. | Guiseppi-Elie A.,Clemson University
Biomaterials | Year: 2010

Electroconductive hydrogels (ECHs) are composite biomaterials that bring together the redox switching and electrical properties of inherently conductive electroactive polymers (CEPs) with the facile small molecule transport, high hydration levels and biocompatibility of cross-linked hydrogels. General methods for the synthesis of electroconductive hydrogels as polymer blends and as polymer co-networks via chemical oxidative, electrochemical and/or a combination of chemical oxidation followed by electrochemical polymerization techniques are reviewed. Specific examples are introduced to illustrate the preparation of electroconductive hydrogels that were synthesized from poly(HEMA)-based hydrogels with polyaniline and from poly(HEMA)-based hydrogels with polypyrrole. The key applications of electroconductive hydrogels; as biorecognition membranes for implantable biosensors, as electro-stimulated drug release devices for programmed delivery, and as the low interfacial impedance layers on neuronal prostheses are highlighted. These applications provide great new horizons for these stimuli responsive, biomimetic polymeric materials. © 2009 Elsevier Ltd. All rights reserved.


Kotanen C.N.,Clemson University | Guiseppi-Elie A.,Clemson University | Guiseppi-Elie A.,ABTECH Scientific Inc.
Biomedical Microdevices | Year: 2013

Continued high morbidity and complications due to trauma related hemorrhage underscores the fact that our understanding of the detailed molecular events of trauma are inadequate to bring life-saving changes to practice. The current state of efficacy and advances in biomedical microdevice technology for trauma diagnostics concerning hemorrhage and hemorrhagic shock was considered with respect to vital signs and metabolic biomarkers. Tachycardia and hypotension are markers of hemorrhagic shock in decompensated trauma patients. Base deficit has been predicative of injury severity at hospital admission. Tissue oxygen saturation has been predicative of onset of multiple organ dysfunction syndrome. Blood potassium levels increase with onset of hemorrhagic shock. Lactate is a surrogate for tissue hypoxia and its clearance predicts mortality. Triage glucose measurements have been shown to be specific in predicting major injuries. No vital sign has yet to be proven effective as an independent predictor of trauma severity. Point of care (POC) devices allow for rapid results, easy sample preparation and processing, small sample volumes, small footprint, multifunctional analysis, and low cost. Advances in the field of in-vivo biosensors has provided a much needed platform by which trauma related metabolites can be monitored easily, rapidly and continuously. Multi-analyte monitoring biosensors have the potential to explore areas still undiscovered in the realm of trauma physiology. © 2013 Springer Science+Business Media New York.

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