Microreactor Array Device

Wiktor, Peter; Brunner, Al; Kahn, Peter; Qiu, Ji; Magee, Mitch; Bian, Xiaofang; Karthikeyan, Kailash; LaBaer, Joshua

doi:10.1038/srep08736

Cited by 19 publications

(20 citation statements)

References 53 publications

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“…The nano-wells were filled with human cell-free expression system (In Vitro Transcription and Translation coupled system; IVTT; Thermo Fisher Scientific) and a custom micro-reactor device was used for the protein expression (23). After sealing the wells with a polystyrene membrane under 200 PSI pressure, we incubated the reactor for 2 h at 30°C for expression and for 0.5 h at 15°C for protein capture, followed by blocking with 5% skim milk in phosphate buffered saline with 0.2% tween 20 (PBST) for 30 min.…”

Section: Active Tuberculosis (Tb)mentioning

confidence: 99%

Identification of Antibody Targets for Tuberculosis Serology using High-Density Nucleic Acid Programmable Protein Arrays

Song

Wallstrom

et al. 2017

Molecular & Cellular Proteomics

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Better and more diverse biomarkers for the development of simple point-of-care tests for active tuberculosis (TB), a clinically heterogeneous disease, are urgently needed. We generated a proteomic Mycobacterium tuberculosis (Mtb) High-Density Nucleic Acid Programmable Protein Array (HD-NAPPA) that used a novel multiplexed strategy for expedited high-throughput screening for antibody responses to the Mtb proteome. We screened sera from HIV uninfected and coinfected TB patients and controls (n ‫؍‬ 120) from the US and South Africa (SA) using the multiplex HD-NAPPA for discovery, followed by deconvolution and validation through single protein HD-NAPPA with biologically independent samples (n ‫؍‬ 124). We verified the top proteins with enzyme-linked immunosorbent assays (ELISA) using the original screening and validation samples (n ‫؍‬ 244) and heretofore untested samples (n ‫؍‬ 41). We identified 8 proteins with TB biomarker value; four (Rv0054, Rv0831c, Rv2031c and Rv0222) of these were previously identified in serology studies, and four (Rv0948c, Rv2853, Rv3405c, Rv3544c) were not known to elicit antibody responses. Using ELISA data, we created classifiers that could discriminate patients' TB status according to geography (

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Section: Active Tuberculosis (Tb)mentioning

confidence: 99%

Identification of Antibody Targets for Tuberculosis Serology using High-Density Nucleic Acid Programmable Protein Arrays

Song

Wallstrom

et al. 2017

Molecular & Cellular Proteomics

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“…Since the cell is compatible with organic electrolytes this device can be used in the future for applications like peptide or nucleotide synthesis that can benefit from the combined effect of the improved pH control and the decrease of importance of the role of the diffusion of reagents [24,25] .…”

Section: Discussionmentioning

confidence: 99%

“…Nano-structuration of electrodes achieved by methods like platinisation [33] can increase the electrochemical surface area and the effective S. were defined using also optical lithography on the epoxy resist (SU-8). WE and CE were platinised following standard recipes [33] However the microfluidic filling of the cell may present some issues due to the small dimensions of the channel and the large area of the chip [25] . To overcome these problems vacuum was applied between the chip and the top glass before the filling.…”

Section: A Design and Fabrication Of The µ-Electrochemical Cellsmentioning

confidence: 99%

Electrochemical Control of pH in Nanoliter Volumes

et al. 2018

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The electrochemical management of the proton concentration in miniaturized dimensions opens the way to control and parallelize multistep chemical reactions, but still it faces many challenges linked to the efficient proton generation and control of their diffusion. Here we present a device operated electrochemically that demonstrates the control of the pH in a cell of ∼140 nL. The device comprises a microfluidic reactor integrated with a pneumatic mechanism that allows the exchange of reagents and the isolation of protons to decrease the effect of their diffusion. We monitored the pH with a fluorescence marker and calculated the final value from the redox currents. We demonstrate a large pH amplitude control from neutral pH values beyond the fluorescence marker range at pH 5. On the basis of the calculations from the Faradaic currents, the minimum pH reached should undergo pH ∼ 0.9. The pH contrast between neutral and acid pH cells can be maintained during periods longer than 15 min with an appropriate design of a diffusion barrier.

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“…After printing, SiNW substrates were blocked using SuperBlock TB (Thermo Scientific, Waltham, MA, USA) for 30 min. The substrates were rinsed, dried, and placed in an airtight chamber with a flexible film above the substrates (micro array device ). Air in the chamber was removed by vacuum from a fluid gap between the substrate and the flexible film.…”

Section: Methodsmentioning

confidence: 99%

Antiviral antibody profiling by high‐density protein arrays

et al. 2015

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Viral infections elicit anti-viral antibodies and have been associated with various chronic diseases. Detection of these antibodies can facilitate diagnosis, treatment of infection and understanding of the mechanisms of virus associated diseases. In this work, we assayed anti-viral antibodies using a novel high density-nucleic acid programmable protein array (HD-NAPPA) platform. Individual viral proteins were expressed in situ directly from plasmids encoding proteins in an array of microscopic reaction chambers. Quality of protein display and serum response was assured by comparing intra- and inter- array correlation within or between printing batches with average correlation coefficients of 0.91 and 0.96, respectively. HD-NAPPA showed higher signal to background (S/B) ratio compared with standard NAPPA on planar glass slides and ELISA. Antibody responses to 761 antigens from 25 different viruses were profiled among patients with juvenile idiopathic arthritis (JIA) and type 1 diabetes (T1D). Common as well as unique antibody reactivity patterns were detected between patients and healthy controls. We believe HD-viral-NAPPA will enable the study of host-pathogen interactions at unprecedented dimensions and elucidate the role of pathogen infections in disease development.

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Microreactor Array Device

Cited by 19 publications

References 53 publications

Identification of Antibody Targets for Tuberculosis Serology using High-Density Nucleic Acid Programmable Protein Arrays

Identification of Antibody Targets for Tuberculosis Serology using High-Density Nucleic Acid Programmable Protein Arrays

Electrochemical Control of pH in Nanoliter Volumes

Antiviral antibody profiling by high‐density protein arrays

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