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Funk C.,Humboldt University of Berlin | Dietrich P.M.,BAM Federal Institute of Materials Research and Testing | Gross T.,BAM Federal Institute of Materials Research and Testing | Min H.,BAM Federal Institute of Materials Research and Testing | And 3 more authors.
Surface and Interface Analysis | Year: 2012

Methods for characterization of epoxy-functionalized substrates used for microarray applications, prepared by silanization with 3- glycidoxypropyltrimethoxysilane, have been developed. Contact angle measurements, X-ray photoelectron spectroscopy, time of flight secondary ion mass spectrometry and fluorescence based methods have been applied to investigate these epoxy-functionalized microarray substrates. The surface density of epoxy-functionalized glass slides was investigated by fluorescence labeling of surface species utilizing Rhodamine 110 as fluorescence probe. Copyright © 2011 John Wiley & Sons, Ltd.

Melamed S.,Hebrew University of Jerusalem | Ceriotti L.,European Commission - Joint Research Center Ispra | Weigel W.,Scienion | Rossi F.,European Commission - Joint Research Center Ispra | And 2 more authors.
Lab on a Chip - Miniaturisation for Chemistry and Biology | Year: 2011

The last decade has witnessed a significant increase in interest in whole-cell biosensors for diverse applications, as well as a rapid and continuous expansion of array technologies. The combination of these two disciplines has yielded the notion of whole-cell array biosensors. We present a potential manifestation of this idea by describing the printing of a whole-cell bacterial bioreporters array. Exploiting natural bacterial tendency to adhere to positively charged abiotic surfaces, we describe immobilization and patterning of bacterial "spots" in the nanolitre volume range by a non-contact robotic printer. We show that the printed Escherichia coli-based sensor bacteria are immobilized on the surface, and retain their viability and biosensing activity for at least 2 months when kept at 4 °C. Immobilization efficiency was improved by manipulating the bacterial genetics (overproducing curli protein), the growth and the printing media (osmotic stress and osmoprotectants) and by a chemical modification of the inanimate surface (self-assembled layers of 3-aminopropyl-triethoxysilane). We suggest that the methodology presented herein may be applicable to the manufacturing of whole-cell sensor arrays for diverse high throughput applications. © 2011 The Royal Society of Chemistry.

Friedrich T.,University of Wurzburg | Rahmann S.,TU Dortmund | Weigel W.,Scienion | Rabsch W.,Robert Koch Institute | And 8 more authors.
BMC Genomics | Year: 2010

Background: The Enterobacteriaceae comprise a large number of clinically relevant species with several individual subspecies. Overlapping virulence-associated gene pools and the high overall genome plasticity often interferes with correct enterobacterial strain typing and risk assessment. Array technology offers a fast, reproducible and standardisable means for bacterial typing and thus provides many advantages for bacterial diagnostics, risk assessment and surveillance. The development of highly discriminative broad-range microbial diagnostic microarrays remains a challenge, because of marked genome plasticity of many bacterial pathogens.Results: We developed a DNA microarray for strain typing and detection of major antimicrobial resistance genes of clinically relevant enterobacteria. For this purpose, we applied a global genome-wide probe selection strategy on 32 available complete enterobacterial genomes combined with a regression model for pathogen classification. The discriminative power of the probe set was further tested in silico on 15 additional complete enterobacterial genome sequences. DNA microarrays based on the selected probes were used to type 92 clinical enterobacterial isolates. Phenotypic tests confirmed the array-based typing results and corroborate that the selected probes allowed correct typing and prediction of major antibiotic resistances of clinically relevant Enterobacteriaceae, including the subspecies level, e.g. the reliable distinction of different E. coli pathotypes.Conclusions: Our results demonstrate that the global probe selection approach based on longest common factor statistics as well as the design of a DNA microarray with a restricted set of discriminative probes enables robust discrimination of different enterobacterial variants and represents a proof of concept that can be adopted for diagnostics of a wide range of microbial pathogens. Our approach circumvents misclassifications arising from the application of virulence markers, which are highly affected by horizontal gene transfer. Moreover, a broad range of pathogens have been covered by an efficient probe set size enabling the design of high-throughput diagnostics. © 2010 Friedrich et al; licensee BioMed Central Ltd.

Siegel G.,Charite - Medical University of Berlin | Siegel G.,Uppsala University | Ermilov E.,Charite - Medical University of Berlin | Knes O.,Swiss Analysis AG | And 2 more authors.
Atherosclerosis | Year: 2014

In a clinical pilot study with eleven metabolic syndrome patients, a simultaneous decrease in hs-CRP from 8.85±4.09 to 4.92±2.51mg/L (-44.4%) (p<0.0436) and HOMA-IR from 3.07±0.63 to 2.60±0.51. mU/L×mg/dL (-15.3%) (p<0.0120) as well as a beneficial change of arteriosclerotic, inflammatory and oxidative stress biomarkers were detected after 2-month treatment with Ginkgo biloba. Furthermore, both IL-6 (-12.9%, p<0.0407) and nanoplaque formation (-14.3%, p<0.0077) were additionally reduced. According to a large clinical trial elucidating the importance of insulin resistance and low-grade systemic inflammation for cardiovascular disease and overall mortality risk, these data might indicate a CVD/total mortality risk reduction. © 2014 Elsevier Ireland Ltd.

Eickhoff H.,Scienion | Malik A.,Microdiscovery GmbH
Advances in Biochemical Engineering/Biotechnology | Year: 2013

A technology for protein microarrays in 96 well microplates has been developed as a widely adoptable platform for multiplexed protein based diagnostics. Procedures for protein microarray manufacturing, immobilization, assay optimization and optical detection methods were developed. Using a clever combination of state of the art technologies, a versatile platform that works with peptides, proteins and antibodies as capture agents in a scalable format for planar arrays is described. © Springer-Verlag Berlin Heidelberg 2013.

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