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Immuno-RAFT Fluorescence Array Sensing for High-Throughput Analysis of CD81 Protein.

Nan Ma1, Yuhui Zhang1, Wei Li1

  • 1School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, Jiangsu 210094, P. R. China.

Analytical Chemistry
|October 20, 2025
PubMed
Summary

A novel fluorescence array using immunosorbent reversible addition-fragmentation chain transfer (immuno-RAFT) offers sensitive detection of CD81 protein for preeclampsia diagnosis. This biosensor method enhances signal amplification for complex clinical samples.

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Clinical Diagnostics

Background:

  • Preeclampsia diagnosis relies on biomarkers like CD81 protein, but traditional methods lack sensitivity for early detection.
  • Enzyme-linked immunosorbent assay (ELISA) is widely used but has limitations in signal amplification for low-concentration analytes.
  • Reversible Addition-Fragmentation chain Transfer (RAFT) polymerization offers robust signal amplification potential for biosensing applications.

Purpose of the Study:

  • To develop a highly sensitive fluorescence array for quantitative detection of CD81 protein using immuno-RAFT signal amplification.
  • To integrate ELISA's operational efficiency with RAFT's amplification capabilities for improved biomarker detection.
  • To validate the method's performance in complex clinical samples, including serum and artificial urine.

Main Methods:

  • Development of a fluorescence array incorporating immunosorbent reversible addition-fragmentation chain transfer (immuno-RAFT) for CD81 detection.
  • Optimization of the RAFT solution with molybdenum disulfide (MoS2) catalyst and piperine to enhance fluorescence signals.
  • Characterization of the fluorescent polymers using particle size analysis and scanning electron microscopy (SEM).

Main Results:

  • The immuno-RAFT fluorescence array demonstrated high sensitivity and quantitative detection of low-concentration CD81 protein.
  • The system showed a strong correlation between fluorescence intensity and protein concentration, with excellent anti-interference capabilities.
  • Formed fluorescent polymers exhibited uniform network structures, confirming the signal amplification mechanism and long-term stability (>95% signal retention after 7 days).

Conclusions:

  • The developed immuno-RAFT fluorescence array sensing method overcomes limitations of traditional techniques for CD81 detection.
  • This advanced biosensor offers practical application for analyzing complex clinical samples and diagnosing preeclampsia.
  • The method shows potential for detecting other protein biomarkers, advancing clinical diagnostics and disease management.