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<p>Raman spectroscopy (RS) is considered a highly promising analytical technique for the rapid and early diagnosis of diseases using biological fluids. However, disease-associated biomarkers are often present in low molecular weight fractions (LMWFs) of serum and can be masked by highly abundant high molecular weight serum proteins, limiting their effective characterization. In the present study, a 100&#xa0;kDa centrifugal ultrafiltration approach was employed to isolate low molecular weight filtrate fractions from whole serum samples of hypopharynx cancer (HPC) patients and healthy controls. The obtained filtrates were analyzed using silver nanoparticles (AgNPs) assisted surface-enhanced Raman spectroscopy (SERS) to investigate biochemical alterations associated with different stages of HPC. Silver nanoparticles exhibiting strong surface plasmon resonance were utilized as enhancing substrates to improve weak Raman signal detection. Distinct spectral variations associated with proteins, nucleic acids, lipids, and other biomolecular constituents were observed between healthy controls and HPC samples. Chemometric techniques including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and receiver operating characteristic (ROC) analysis were applied for spectral discrimination and classification. The PLS-DA model demonstrated 79% classification accuracy with 81% sensitivity, 76% specificity, and an AUC value of 0.73 for differentiating HPC samples from healthy controls. The developed filtration-assisted SERS platform improved the detection of low molecular weight serum biomarkers by minimizing spectral interference from highly abundant serum proteins, thereby enhancing disease-related biochemical information. The findings support the feasibility of ultrafiltration-assisted SERS combined with chemometric analysis for the stage-wise characterization of hypopharynx cancer. However, larger multicenter studies and external validation are required to further establish the clinical utility and generalizability of the proposed approach.</p>

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Surface-enhanced Raman spectroscopy for the characterization of filtrate portions of blood serum samples of hypopharynx cancer patients using 100 kDa filtration devices and AgNPs as SERS substrate

  • 期刊论文

  • 发表日期：

    2026年7月23日

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  • Usman Munawer,
  • Nosheen Rashid,
  • Muhammad Irfan Majeed,
  • Haq Nawaz,
  • Hafiza Elbadie Ahmed,
  • Muhammad Zeeshan Majeed,
  • Javeria Ameen,
  • Muhammad Roman,
  • Muhammad Shahid,
  • Zaid Aslam,
  • Misbah Naeem,
  • Fozia Sarfraz,
  • Aqsa Nawaz,
  • Muhammad Imran

摘要

Raman spectroscopy (RS) is considered a highly promising analytical technique for the rapid and early diagnosis of diseases using biological fluids. However, disease-associated biomarkers are often present in low molecular weight fractions (LMWFs) of serum and can be masked by highly abundant high molecular weight serum proteins, limiting their effective characterization. In the present study, a 100 kDa centrifugal ultrafiltration approach was employed to isolate low molecular weight filtrate fractions from whole serum samples of hypopharynx cancer (HPC) patients and healthy controls. The obtained filtrates were analyzed using silver nanoparticles (AgNPs) assisted surface-enhanced Raman spectroscopy (SERS) to investigate biochemical alterations associated with different stages of HPC. Silver nanoparticles exhibiting strong surface plasmon resonance were utilized as enhancing substrates to improve weak Raman signal detection. Distinct spectral variations associated with proteins, nucleic acids, lipids, and other biomolecular constituents were observed between healthy controls and HPC samples. Chemometric techniques including principal component analysis (PCA), partial least squares-discriminant analysis (PLS-DA), and receiver operating characteristic (ROC) analysis were applied for spectral discrimination and classification. The PLS-DA model demonstrated 79% classification accuracy with 81% sensitivity, 76% specificity, and an AUC value of 0.73 for differentiating HPC samples from healthy controls. The developed filtration-assisted SERS platform improved the detection of low molecular weight serum biomarkers by minimizing spectral interference from highly abundant serum proteins, thereby enhancing disease-related biochemical information. The findings support the feasibility of ultrafiltration-assisted SERS combined with chemometric analysis for the stage-wise characterization of hypopharynx cancer. However, larger multicenter studies and external validation are required to further establish the clinical utility and generalizability of the proposed approach.