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From the third issue of 2017, Microarrays has changed its name to High-Throughput.

Open AccessArticle
Microarrays 2014, 3(4), 282-301; doi:10.3390/microarrays3040282

Fabrication of Homogeneous High-Density Antibody Microarrays for Cytokine Detection

1
Materials Science Laboratory, Sony Deutschland GmbH, Hedelfinger Strasse 61, 70327 Stuttgart, Germany
2
Institute of Electronic and Sensor Materials, Technische Universität Bergakademie Freiberg, Gustav-Zeuner-Strasse 3, 09599 Freiberg, Germany
*
Author to whom correspondence should be addressed.
Received: 6 June 2014 / Revised: 15 November 2014 / Accepted: 1 December 2014 / Published: 9 December 2014
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Abstract

Cytokine proteins are known as biomarker molecules, characteristic of a disease or specific body condition. Monitoring of the cytokine pattern in body fluids can contribute to the diagnosis of diseases. Here we report on the development of an array comprised of different anti-cytokine antibodies on an activated solid support coupled with a fluorescence readout mechanism. Optimization of the array preparation was done in regard of spot homogeneity and spot size. The proinflammatory cytokines Tumor Necrosis Factor alpha (TNFα) and Interleukin 6 (IL-6) were chosen as the first targets of interest. First, the solid support for covalent antibody immobilization and an adequate fluorescent label were selected. Three differently functionalized glass substrates for spotting were compared: amine and epoxy, both having a two-dimensional structure, and the NHS functionalized hydrogel (NHS-3D). The NHS-hydrogel functionalization of the substrate was best suited to antibody immobilization. Then, the optimization of plotting parameters and geometry as well as buffer media were investigated, considering the ambient analyte theory of Roger Ekins. As a first step towards real sample studies, a proof of principle of cytokine detection has been established. View Full-Text
Keywords: microarray; protein; antibody; cytokines; spotting; hydrogel; biosensor; fluorescence readout microarray; protein; antibody; cytokines; spotting; hydrogel; biosensor; fluorescence readout
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Hospach, I.; Joseph, Y.; Mai, M.K.; Krasteva, N.; Nelles, G. Fabrication of Homogeneous High-Density Antibody Microarrays for Cytokine Detection. Microarrays 2014, 3, 282-301.

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