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An 8 minute colorimetric paper-based reverse phase vertical flow serum microarray for screening of hyper IgE syndrome
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.ORCID iD: 0000-0003-0344-8049
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology.
2015 (English)In: The Analyst, ISSN 0003-2654, E-ISSN 1364-5528, Vol. 140, no 21, 7327-7334 p.Article in journal (Refereed) PublishedText
Abstract [en]

Reverse phase microarrays are useful tools for affinity-based detection in hundreds of samples simultaneously. However, current methods typically require long assay times and fluorescent detection. Here we describe a paper-based Vertical Flow Microarray (VFM) assay as a rapid 8-minute colorimetric alternative for reverse phase microarray analysis. The VFM platform was optimized for detection of IgE with a detection limit of 1.9 μg mL-1 in whole serum. Optimized conditions were then used to screen 113 serum samples simultaneously for hyper IgE syndrome (hIgE), a rare primary immunodeficiency characterized by elevated levels of IgE. The same set of samples were then analysed with a conventional planar microarray with fluorescent detection for head-to-head testing. Both assays found elevated levels in three out of four hIgE patient samples, whereas no control samples displayed elevated levels in either method. The comparison experiments showed a good correlation between the two assays, as determined from a linear correlation study (Pearson's r = 0.76). Further, the assay-time reduction and reproducibility (intra assay CV = 12.4 ± 4.11%) demonstrate the applicability of the VFM platform for high throughput reverse phase screening.

Place, publisher, year, edition, pages
Royal Society of Chemistry, 2015. Vol. 140, no 21, 7327-7334 p.
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:kth:diva-181205DOI: 10.1039/c5an01013fISI: 000362795100031ScopusID: 2-s2.0-84944096876OAI: oai:DiVA.org:kth-181205DiVA: diva2:902850
Funder
Swedish Research Council
Note

QC 20160212

Available from: 2016-02-12 Created: 2016-01-29 Last updated: 2016-02-12Bibliographically approved

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Reuterswärd, PhilippaGantelius, JesperAndersson Svahn, Helene
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