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<p>Microfluidic multiplexed immunosensors enable the simultaneous detection of multiple cancer biomarkers. This study highlights a patented technology (US-11,828,747-B2) integrating surface-enhanced Raman spectroscopy (SERS) with a plasmonic-active nanostructure thin film (PANTF) on a printed circuit board (PCB), housed within a microfluidic device. Designed for breast cancer (BC) diagnosis, it allows rapid, non-destructive, and parallel quantification of biomarkers in blood serum (BCS). Detection uses monoclonal antibody (mAb)-functionalized gold nanourchins (GNUs) on dual sensing regions. A total of 46 serum samples (92 data points) were tested for HER-II and CA 15 − 3. The system demonstrated a SERS enhancement factor (EF) of ~ 0.35 × 10⁵ using Rhodamine 6G. Calibration with HER-II (10–100 ng/mL) and CA 15 − 3 (10–100 U/mL) showed linear responses (R² = 0.942 and 0.8, respectively). Measurements of unknowns were performed at 1 µL/min over 68&#xa0;min, with 43&#xa0;min for biomarker interaction. Raman spectra were recorded at active zones and analyzed using SpectraView (SV), a custom Python-based tool. Data preprocessing included filtering (<i>SciPy’s filtfilt</i>) and baseline correction using the Improved Asymmetric Least Squares (<i>IASLS</i>) algorithm (<i>pybaselines.Whittaker</i>). Fourier cross-correlation (FCC) showed stronger signal consistency for HER-II. Partial Least Squares (PLS) regression, a dimensionality reduction technique, enabled clear discrimination between HER-II and CA 15 − 3, with classification accuracy reaching 1.0. Cancer staging based on these biomarkers yielded an overall accuracy of 0.90, indicating effective classification regardless of biomarker type. Individual accuracies were 0.93 for HER-II and 0.80 for CA 15 − 3.</p> Graphical Abstract <p></p>

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Label-Free Multiplex SERS Microfluidic Immunoassay of Breast Cancer Biomarkers in Serum Using Directionally Conjugated mAb-Gold Nanourchins on a PCB-Thin Gold Film Platform

  • 期刊论文

  • 发表日期：

    2025年12月23日

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  • Mohammad E. Khosroshahi,
  • Gayathri Senthilchelvan,
  • Adam Pezeshki,
  • Victor Oyebolu

摘要

Microfluidic multiplexed immunosensors enable the simultaneous detection of multiple cancer biomarkers. This study highlights a patented technology (US-11,828,747-B2) integrating surface-enhanced Raman spectroscopy (SERS) with a plasmonic-active nanostructure thin film (PANTF) on a printed circuit board (PCB), housed within a microfluidic device. Designed for breast cancer (BC) diagnosis, it allows rapid, non-destructive, and parallel quantification of biomarkers in blood serum (BCS). Detection uses monoclonal antibody (mAb)-functionalized gold nanourchins (GNUs) on dual sensing regions. A total of 46 serum samples (92 data points) were tested for HER-II and CA 15 − 3. The system demonstrated a SERS enhancement factor (EF) of ~ 0.35 × 10⁵ using Rhodamine 6G. Calibration with HER-II (10–100 ng/mL) and CA 15 − 3 (10–100 U/mL) showed linear responses (R² = 0.942 and 0.8, respectively). Measurements of unknowns were performed at 1 µL/min over 68 min, with 43 min for biomarker interaction. Raman spectra were recorded at active zones and analyzed using SpectraView (SV), a custom Python-based tool. Data preprocessing included filtering (SciPy’s filtfilt) and baseline correction using the Improved Asymmetric Least Squares (IASLS) algorithm (pybaselines.Whittaker). Fourier cross-correlation (FCC) showed stronger signal consistency for HER-II. Partial Least Squares (PLS) regression, a dimensionality reduction technique, enabled clear discrimination between HER-II and CA 15 − 3, with classification accuracy reaching 1.0. Cancer staging based on these biomarkers yielded an overall accuracy of 0.90, indicating effective classification regardless of biomarker type. Individual accuracies were 0.93 for HER-II and 0.80 for CA 15 − 3.

Graphical Abstract