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Flow cytometry-based validation of soluble biomarker detection.

Yiting Tang1, Xiang Wu1,2, Huating Zhang1

  • 1Department of Laboratory Medicine, Huashan Hospital, Fudan University, Shanghai, China.

Practical Laboratory Medicine
|November 28, 2025
PubMed
Summary

This study validates a multiplex flow cytometry assay for simultaneously measuring soluble immune markers (sCD25, sCD40L, sCD130, sTREM-1) in serum. The sensitive and reproducible assay shows potential for lung cancer diagnosis and treatment monitoring.

Keywords:
BD FACSLyricFlow cytometryImmune responseSoluble markerssCD130sCD25sCD40LsTREM-1

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Area of Science:

  • Immunology
  • Biotechnology
  • Clinical Diagnostics

Background:

  • Accurate measurement of soluble immune markers like sCD25, sCD40L, sCD130, and sTREM-1 is vital for understanding immune responses and inflammation.
  • These markers are implicated in various inflammatory conditions and have been linked to diseases such as lung cancer.

Purpose of the Study:

  • To validate a multiplex flow cytometry assay for the simultaneous quantification of four key soluble immune markers in serum.
  • To assess the assay's sensitivity, reproducibility, and utility in distinguishing between lung cancer patients and healthy individuals.

Main Methods:

  • Utilized a bead-based immunoassay kit compatible with the BD FACSLyric flow cytometer.
  • Optimized sample preparation, bead conjugation, and assay parameters for precise detection.
  • Acquired data on BD FACSLyric, recording a minimum of 10,000 events per sample for robust analysis.

Main Results:

  • The validated assay demonstrated low limits of detection (LOD) for sCD25 (9.77 pg/ml), sCD40L (43.95 pg/ml), sCD130 (219.73 pg/ml), and sTREM-1 (12.21 pg/ml).
  • High reproducibility was confirmed through intra-assay and inter-assay variability assessments.
  • Significant differences in marker levels were observed between lung cancer patients and healthy individuals, indicating diagnostic potential.

Conclusions:

  • The validated multiplex assay offers a sensitive, reproducible, and efficient method for quantifying multiple soluble immune markers simultaneously.
  • This assay is a valuable tool for both research and clinical applications, particularly in aiding the diagnosis and treatment of lung cancer.
  • Future developments may involve expanding the assay to include additional markers or adapting it for different sample types.