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Biosensors 2018, 8(1), 10; https://doi.org/10.3390/bios8010010

Investigating Colorimetric Protein Array Assay Schemes for Detection of Recurrence of Bladder Cancer

Center for Health & Bioresources, AIT Austrian Institute of Technology, Konrad Lorenz Straße 24, Tulln 3430, Austria
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Received: 22 November 2017 / Revised: 13 January 2018 / Accepted: 13 January 2018 / Published: 24 January 2018
(This article belongs to the Special Issue Biomarkers)
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Abstract

A colorimetric microarray for the multiplexed detection of recurrence of bladder cancer including protein markers interleukin-8 (IL8), decorin (DCN), and vascular endothelial growth factor (VEGF) was established to enable easy and cheap read-out by a simple office scanner paving the way for quick therapy monitoring at doctors’ offices. The chip is based on the principle of a sandwich immunoassay and was optimized prior to multiplexing using IL8 as a model marker. Six different colorimetric assay formats were evaluated using a detection antibody (dAB) labeled with (I) gold (Au) nanoparticles (NPs), (II) carbon NPs, (III) oxidized carbon NPs, and a biotinylated dAB in combination with (IV) neutravidin–carbon, (V) streptavidin (strp)–gold, and (VI) strp–horseradish peroxidase (HRP). Assay Format (III) worked best for NP-based detection and showed a low background while the enzymatic approach, using 3,3′,5,5′-tetramethylbenzidine (TMB) substrate, led to the most intense signals with good reproducibility. Both assay formats showed consistent spot morphology as well as detection limits lower than 15 ng/L IL8 and were thus applied for the multiplexed detection of IL8, DCN, and VEGF in synthetic urine. Colorimetric detection in urine (1:3) yields reaction signals and measurement ranges well comparable with detection in the assay buffer, as well as excellent data reproducibility as indicated by the coefficient of variation (CV 5–9%). View Full-Text
Keywords: bladder cancer; IL8; VEGF; DCN; colorimetric protein array; carbon nanoparticles bladder cancer; IL8; VEGF; DCN; colorimetric protein array; carbon nanoparticles
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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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Gogalic, S.; Sauer, U.; Doppler, S.; Preininger, C. Investigating Colorimetric Protein Array Assay Schemes for Detection of Recurrence of Bladder Cancer. Biosensors 2018, 8, 10.

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