Title: Fluorescent vesicles for signal amplification in reverse phase protein microarray assays
Authors : Bally, Marta
Syed, Shahida
Binkert, Andreas
Kauffmann, Ekkehard
Ehrat, Markus
Vörös, Janos
Published in : Analytical Biochemistry
Volume(Issue) : 416
Issue : 2
Pages : 145
Pages to: 151
Publisher / Ed. Institution : Elsevier
Issue Date: 2011
License (according to publishing contract) : Licence according to publishing contract
Type of review: Peer review (Publication)
Language : English
Subjects : Animal; Antibodies; Fluorescent Dye; Immunoassay; Immunoglobulin G; Lipid Bilayer; Microscopy, Confocal; Protein Array Analysis; Rabbit; Rhodamines
Subject (DDC) : 570: Biology
615: Pharmacology and therapeutics
Abstract: Developments in microarray technology promise to lead to great advancements in the biomedical and biological field. However, implementation of these analytical tools often relies on signal amplification strategies that are essential to reach the sensitivity levels required for a variety of biological applications. This is true especially for reverse phase arrays where a complex biological sample is directly immobilized on the chip. We present a simple and generic method for signal amplification based on the use of antibody-tagged fluorescent vesicles as labels for signal generation. To assess the gain in assay sensitivity, we performed a model assay for the detection of rabbit immunoglobulin G (IgG) and compared the limit of detection (LOD) of the vesicle assay with the LOD of a conventional assay performed with fluorescent reporter molecules. We evaluated the improvements for two fluorescence-based transduction setups: a high-sensitivity microarray reader (ZeptoREADER) and a conventional confocal scanner. In all cases, our strategy led to an increase in sensitivity. However, gain in sensitivity widely depended on the type of illumination; whereas an approximately 2-fold increase in sensitivity was observed for readout based on evanescent field illumination, the contribution was as high as more than 200-fold for confocal scanning.
Departement: School of Engineering
Publication type: Article in scientific Journal
DOI : 10.1016/j.ab.2011.05.030
ISSN: 0003-2697
URI: https://digitalcollection.zhaw.ch/handle/11475/1734
Appears in Collections:Publikationen School of Engineering

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