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

Selma Gogalic1, Ursula Sauer2, Sara Doppler3

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A new colorimetric microarray enables multiplexed detection of bladder cancer recurrence markers, including interleukin-8 (IL8), decorin (DCN), and vascular endothelial growth factor (VEGF). This cost-effective method allows for rapid therapy monitoring in doctors' offices.

Keywords:
DCNIL8VEGFbladder cancercarbon nanoparticlescolorimetric protein array

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Oncology Diagnostics

Background:

  • Bladder cancer recurrence monitoring requires accessible and cost-effective diagnostic tools.
  • Current methods for detecting protein biomarkers like IL8, DCN, and VEGF can be complex and time-consuming.
  • Point-of-care diagnostics are crucial for timely therapeutic adjustments.

Purpose of the Study:

  • To develop a colorimetric microarray for multiplexed detection of bladder cancer recurrence markers.
  • To establish an easy and affordable readout method using a standard office scanner for rapid therapy monitoring.
  • To optimize assay formats for sensitive and reproducible detection of IL8, DCN, and VEGF.

Main Methods:

  • A sandwich immunoassay principle was employed on a microarray chip.
  • Six different colorimetric assay formats were evaluated, including nanoparticle and enzymatic detection methods.
  • Interleukin-8 (IL8) was used as a model marker for initial optimization before multiplexing.

Main Results:

  • Assay Format III (oxidized carbon NPs) and an enzymatic approach (streptavidin-horseradish peroxidase) showed optimal performance.
  • Detection limits for IL8 were below 15 ng/L with good reproducibility (CV 5-9%) in synthetic urine.
  • The developed microarray successfully detected IL8, DCN, and VEGF in urine samples, comparable to assay buffer detection.

Conclusions:

  • A novel colorimetric microarray enables multiplexed detection of key bladder cancer recurrence markers.
  • The assay is suitable for rapid, cost-effective monitoring in clinical settings, facilitating timely treatment decisions.
  • The developed method demonstrates high sensitivity, reproducibility, and applicability in a clinically relevant matrix (urine).