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Colorimetric paper-based sarcosine assay with improved sensitivity.

Mayu Masumoto1, Sera Ohta1, Misaki Nakagawa1

  • 1Department of Applied Chemistry, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku-ku, Yokohama, 223-8522, Japan.

Analytical and Bioanalytical Chemistry
|October 18, 2021
PubMed
Summary

This study presents a novel paper-based assay for detecting sarcosine, a prostate cancer biomarker. The assay uses enzymes on paper to detect sarcosine, achieving a low detection limit for improved diagnostics.

Keywords:
Artificial urineCationically charged polyamineEnzymatic sarcosine assayHydrogen peroxideProstate cancer biomarkerSarcosine oxidase

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Area of Science:

  • Biomarker detection
  • Point-of-care diagnostics
  • Enzyme-based assays

Background:

  • Prostate cancer diagnosis relies on biomarkers like sarcosine.
  • Existing detection methods can be complex and costly.
  • A simple, accessible detection method for sarcosine is needed.

Purpose of the Study:

  • To develop a paper-based colorimetric assay for sarcosine detection.
  • To improve the sensitivity and limit of detection for sarcosine.
  • To validate the assay's performance in artificial urine.

Main Methods:

  • A bienzymatic colorimetric assay using sarcosine oxidase (SOx) and horseradish peroxidase (HRP) on filter paper.
  • Modification of paper with poly(allylamine hydrochloride) (PAH) to enhance sensitivity.
  • Inclusion of pH-buffer components for on-device sample adjustment.
  • Testing in artificial urine matrix and assessment of selectivity and stability.

Main Results:

  • Achieved a low limit of detection (LOD) of 0.6 μM for sarcosine on modified paper, significantly lower than unmodified paper (12.6 μM).
  • Demonstrated a linear response to sarcosine between 0 and 10 μM.
  • Successfully eliminated bicarbonate inhibition and achieved an LOD of 2.5 μM in artificial urine, below the diagnostic threshold.
  • Confirmed good selectivity over 20 amino acids and storage stability for 50 days at -20°C.

Conclusions:

  • The paper-based assay offers a sensitive and stable method for sarcosine detection.
  • The assay's performance in artificial urine meets diagnostic requirements.
  • This approach holds promise for simple, low-cost prostate cancer screening.