Fluorescent vesicles for signal amplification in reverse phase protein microarray assays

Bally, Marta; Syed, Shahida; Binkert, Andreas; Kauffmann, Ekkehard; Ehrat, Markus; Vörös, Janos

doi:10.1016/j.ab.2011.05.030

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
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
DOI:	10.1016/j.ab.2011.05.030
Published in:	Analytical Biochemistry
Volume(Issue):	416
Issue:	2
Page(s):	145
Pages to:	151
Issue Date:	2011
Publisher / Ed. Institution:	Elsevier
ISSN:	0003-2697 1096-0309
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.
URI:	https://digitalcollection.zhaw.ch/handle/11475/1734
Fulltext version:	Published version
License (according to publishing contract):	Licence according to publishing contract
Departement:	School of Engineering
Appears in collections:	Publikationen School of Engineering

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Bally, M., Syed, S., Binkert, A., Kauffmann, E., Ehrat, M., & Vörös, J. (2011). Fluorescent vesicles for signal amplification in reverse phase protein microarray assays. Analytical Biochemistry, 416(2), 145–151. https://doi.org/10.1016/j.ab.2011.05.030

Bally, M. et al. (2011) ‘Fluorescent vesicles for signal amplification in reverse phase protein microarray assays’, Analytical Biochemistry, 416(2), pp. 145–151. Available at: https://doi.org/10.1016/j.ab.2011.05.030.

M. Bally, S. Syed, A. Binkert, E. Kauffmann, M. Ehrat, and J. Vörös, “Fluorescent vesicles for signal amplification in reverse phase protein microarray assays,” Analytical Biochemistry, vol. 416, no. 2, pp. 145–151, 2011, doi: 10.1016/j.ab.2011.05.030.

BALLY, Marta, Shahida SYED, Andreas BINKERT, Ekkehard KAUFFMANN, Markus EHRAT und Janos VÖRÖS, 2011. Fluorescent vesicles for signal amplification in reverse phase protein microarray assays. Analytical Biochemistry. 2011. Bd. 416, Nr. 2, S. 145–151. DOI 10.1016/j.ab.2011.05.030

Bally, Marta, Shahida Syed, Andreas Binkert, Ekkehard Kauffmann, Markus Ehrat, and Janos Vörös. 2011. “Fluorescent Vesicles for Signal Amplification in Reverse Phase Protein Microarray Assays.” Analytical Biochemistry 416 (2): 145–51. https://doi.org/10.1016/j.ab.2011.05.030.

Bally, Marta, et al. “Fluorescent Vesicles for Signal Amplification in Reverse Phase Protein Microarray Assays.” Analytical Biochemistry, vol. 416, no. 2, 2011, pp. 145–51, https://doi.org/10.1016/j.ab.2011.05.030.