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Updated: Sep 19, 2025

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DNA Hybridization-Accelerated Programmable Immuno-Aggregation of Microbeads Enabling Mix-and-Read Flow Cytometric

Junyue Sun1, Xinrui Fei1, Wenjiao Fan1

  • 1Key Laboratory of Applied Surface and Colloid Chemistry, Ministry of Education; Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province; School of Chemistry & Chemical Engineering, Shaanxi Normal University, Xi'an 710119, Shaanxi Province, P. R. China.

Analytical Chemistry
|June 17, 2025
PubMed
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This summary is machine-generated.

This study introduces a novel DNA hybridization method to improve microbead aggregation for sensitive protein biomarker detection. This approach enhances efficiency and enables precise, multiplexed analysis using flow cytometry.

Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Biotechnology

Background:

  • Microbead (MB) aggregation immunoassays offer label-free protein biomarker analysis but face challenges in low aggregation efficiency and precise measurement.
  • Steric hindrance and bead weight limit target-actuated aggregation, hindering practical applications.

Purpose of the Study:

  • To develop a novel, highly sensitive, and precise method for protein biomarker detection using microbead aggregation.
  • To overcome limitations of existing microbead immunoassays through enhanced aggregation efficiency and accurate quantification.

Main Methods:

  • Proposed a new mechanism of metastable DNA hybridization-accelerated programmable immuno-aggregation of microbeads (MBs).
  • Utilized auxiliary metastable DNA hybridization and magnetic facilitation to boost immunoreaction efficiency between two types of MBs.

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  • Employed flow cytometry for precise interrogation of light scattering and fluorescence information of individual MB aggregates and monomers.

    • Main Results:

    • Achieved an approximately 200-fold increase in detection sensitivity due to enhanced immunoreaction efficiency.
    • Enabled distinct discrimination and precise quantification of MB aggregates from MB monomers using flow cytometry.
    • Demonstrated multiplexed analysis of protein targets by coencoding MBs with fluorescent color and intensity.

    Conclusions:

    • The proposed metastable DNA hybridization-accelerated immuno-aggregation method significantly enhances protein biomarker detection sensitivity and precision.
    • The developed technique allows for facile mix-and-read and flow cytometric detection, overcoming previous limitations.
    • This approach shows great potential for diverse biomedical applications, including sensitive and multiplexed protein analysis.