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Spatiotemporal mapping of concentration polarization induced pH changes at nanoconstrictions

ACS Nano

Mai, Junyu; Miller, Hanna; Hatch, Anson H.

Under an applied electric field, concentration polarization (CP) arises from ion permselectivity of most nanoporous materials and biological ion channels. We present novel methods to quantitatively assess CP-induced spatiotemporal changes of pH that may significantly impact transport dynamics, device functionality, and physicochemical properties of molecular analytes in devices with nanofluidic constrictions. We measured pH fluctuations of >1.5 pH units and changes extending over 100's of micrometers from nanoconstrictions. The degree of change depends on key system parameters including buffer composition, surface charge, and strength of electric field. The results highlight the importance of neglected contributions of pH changes, and the approach can aid characterization and manipulation of mass transport in nanofluidic systems. © 2012 American Chemical Society.

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Ultrasensitive microanalytical diagnostic methods for rickettsial pathogens

Hatch, Anson H.

A strategic CRADA was established between Sandia National Laboratories (SNL) and the University of Texas Medical Branch (UTMB) at Galveston to address pressing needs for US protection against biological weapons of mass destruction (WMD) and emerging infectious diseases. The combination of unique expertise and facilities at UTMB and SNL enabled interdisciplinary research efforts in the development of rapid and accurate diagnostic methods for early detection of trace priority pathogen levels. Outstanding postdoctoral students were also trained at both institutions to help enable the next generation of scientists to tackle the challenging interdisciplinary problems in the area of biodefense and emerging infectious diseases. Novel approaches to diagnostics were developed and the both the speed of assays as well as the detection sensitivity were improved by over an order of magnitude compared to traditional methods. This is a significant step toward more timely and specific detection of dangerous infections. We developed in situ polymerized porous polymer monoliths that can be used as (1) size exclusion elements for capture and processing of rickettsial infected cells from a sample, (2) photopatternable framework for grafting high densities of functionalized antibodies/fluorescent particles using novel monolith chemistry. Grafting affinity reagents specific to rickettsial particles enables rapid, ultra-sensitive assays by overcoming transport limitations of traditional planar assay approaches. We have selectively trapped particles and bacteria at the cell trap and have also detected picomolar mouse IL-6 captured with only 20 minutes total incubation times using the densely patterned monolith framework. As predicted, the monolith exhibits >10x improvements in both capture speed and capture density compared to traditional planar approaches. The most significant advancements as part of this CRADA is the optimization of techniques allowing the detection of <10 rickettsial cells in a whole blood sample. This detection limit is over 2 orders of magnitude more sensitive that previously reported methods and overcomes a key hurdle in ability to sense dangerous infections before they are too late to treat and contain. We also showed that in the new format, cross-reactivity with interfering species is reduced thereby increasing the specificity of such tests. Promising options to treat whole blood and avoid clogging and non-specific fouling of sensors were also developed.

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A fully-integrated aptamer-based affinity assay platform for monitoring astronaut health in space

14th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2010, MicroTAS 2010

Sommer, G.J.; Hecht, A.H.; Durland, R.H.; Yang, X.; Singh, Anup K.; Hatch, Anson H.

Here we demonstrate the suitability of robust nucleic acid affinity reagents in an integrated point-of-care diagnostic platform for monitoring proteomic biomarkers indicative of astronaut health in spaceflight applications. A model thioaptamer[1] targeting nuclear factor-kappa B (NF-KB) is evaluated in an on-chip electrophoretic gel-shift assay for human serum. Key steps of i) mixing sample with the aptamer, ii) buffer exchange, and iii) preconcentration of sample were successfully integrated upstream of fluorescence-based detection. Challenges due to i) nonspecific interactions with serum, and ii) preconcentration at a nanoporous membrane are discussed and successfully resolved to yield a robust, rapid, and fully-integrated diagnostic system.

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An ultra-sensitive microfluidic immunoassay using living radical polymerization and porous polymer monoliths

14th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2010, MicroTAS 2010

Abhyankar, Vinay V.; Singh, Anup K.; Hatch, Anson H.

We present a platform that combines patterned photopolymerized polymer monoliths with living radical polymerization (LRP) to develop a low cost microfluidic based immunoassay capable of sensitive (low to sub pM) and rapid (<30 minute) detection of protein in 100 μL sample. The introduction of LRP functionality to the porous monolith allows one step grafting of functionalized affinity probes from the monolith surface while the composition of the hydrophilic graft chain reduces non-specific interactions and helps to significantly improve the limit of detection.

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Microscale isoelectric fractionation using immobilized ph-specific membranes for multi-dimensional analysis

14th International Conference on Miniaturized Systems for Chemistry and Life Sciences 2010, MicroTAS 2010

Mai, Junyu M.; Sommer, G.J.; Hatch, Anson H.

We report on advancements of our microscale isoelectric fractionation (μIEFr) methodology for fast on-chip separation and concentration of proteins based on their isoelectric points (pI). We establish that proteins can be fractionated depending on posttranslational modifications into different pH specific bins, from where they can be efficiently transferred to downstream membranes for additional processing and analysis. This technology can enable on-chip multidimensional glycoproteomics analysis, as a new approach to expedite biomarker identification and verification.

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Biotechnology development for biomedical applications

Rempe, Susan R.; Rogers, David M.; Buerger, Stephen B.; Kuehl, Michael K.; Hatch, Anson H.; Abhyankar, Vinay V.; Mai, Junyu M.; Dirk, Shawn M.; Brozik, Susan M.; De Sapio, Vincent D.; Schoeniger, Joseph S.

Sandia's scientific and engineering expertise in the fields of computational biology, high-performance prosthetic limbs, biodetection, and bioinformatics has been applied to specific problems at the forefront of cancer research. Molecular modeling was employed to design stable mutations of the enzyme L-asparaginase with improved selectivity for asparagine over other amino acids with the potential for improved cancer chemotherapy. New electrospun polymer composites with improved electrical conductivity and mechanical compliance have been demonstrated with the promise of direct interfacing between the peripheral nervous system and the control electronics of advanced prosthetics. The capture of rare circulating tumor cells has been demonstrated on a microfluidic chip produced with a versatile fabrication processes capable of integration with existing lab-on-a-chip and biosensor technology. And software tools have been developed to increase the calculation speed of clustered heat maps for the display of relationships in large arrays of protein data. All these projects were carried out in collaboration with researchers at the University of Texas M. D. Anderson Cancer Center in Houston, TX.

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Microscale isoelectric fractionation using immobilized pH-specific membranes for multi-dimensional analysis

Sommer, G.J.; Hatch, Anson H.

We report on advancements of our microscale isoelectric fractionation ({mu}IEFr) methodology for fast on-chip separation and concentration of proteins based on their isoelectric points (pI). We establish that proteins can be fractionated depending on posttranslational modifications into different pH specific bins, from where they can be efficiently transferred to downstream membranes for additional processing and analysis. This technology can enable on-chip multidimensional glycoproteomics analysis, as a new approach to expedite biomarker identification and verification.

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Ultra-rapid sample preconcentration under slant field using high-aspect-ratio nanoporous membranes

12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference

Wang, Ying C.; Singh, Anup K.; Hatch, Anson H.

We describe a novel approach to fabricate high-aspect-ratio membranes in microchannels by direct laser scanning, and demonstrate >10-fold improvement in sample preconcentration speed by achieving lower fM detection of proteins within 5 minutes. The integrated device can be used for continuous sample preparation, injection, preconcentration, and biochemical binding/reaction applications. © 2008 CBMS.

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Precise microscale polymer gradients applied to isoelectric focusing and pore limit electrophoresis

12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference

Sommer, G.J.; Singh, Anup K.; Hatch, Anson H.

We demonstrate the power of our technique for establishing and immobilizing well-defined polymer gradients in microchannels by fabricating two miniaturized analytical platforms: microscale immobilized pH gradients (μIPGs) for rapid and high resolution isoelectric focusing (IEF) applications, and polyacrylamide porosity gradients to achieve microscale pore limit electrophoresis (μPLE) in which species are separated based on molecular size by driving them toward the pore size at which migration ceases. Both separation techniques represent the first microscale implementation of their respective methodologies.

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38 Results
38 Results