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Related Experiment Video

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Flow Cytometry-Based Rapid Assay for Antigen Specific Antibody Relative Affinity in SRBC-Immunized Mouse Models.

Chunli Sun1,2, Yuan Jiang1,2, Shujun Liu1,2

  • 1Shanghai Institute of Immunology, Faculty of Basic Medicine, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.

International Journal of Molecular Sciences
|May 7, 2025
PubMed
Summary

This study introduces a novel flow cytometry method to accurately measure sheep red blood cell-specific antibody affinity. This technique aids research in humoral immunity, vaccine response, and immunotoxicity testing.

Keywords:
Src homology 2-containing inositol 5-phosphatase 1 (SHIP-1)T cell-dependent antibody response (TDAR)antibody affinity maturation (AAM)flow cytometryimmunomodulationimmunotoxicity testingsheep red blood cells (SRBC)

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Area of Science:

  • Immunology
  • Biotechnology

Background:

  • Sheep red blood cells (SRBC) are classical T-cell dependent (TD) antigens widely used in immunology research.
  • SRBC are cost-effective and elicit robust antibody responses, making them valuable for studying humoral immunity, vaccine development, and bioactive agent testing.
  • Accurate detection of SRBC-specific antibody affinity levels in animal models is currently challenging.

Purpose of the Study:

  • To establish a rapid and accurate detection system for assessing relative SRBC-specific antibody affinity levels.
  • To provide a technological solution for research involving humoral immunity and antibody responses.

Main Methods:

  • Development of a flow cytometry-based detection system.
  • Utilizing SRBC-immunized mouse models and their humoral samples (serum, tissue fluid).
  • Validation using affinity maturation-deficient mice to confirm method's ability to distinguish antibody affinity levels.

Main Results:

  • A flow cytometry system was successfully established for quick and accurate assessment of SRBC-specific antibody relative affinity.
  • The method demonstrated effectiveness in distinguishing antibody affinity levels between different samples, as validated in affinity maturation-deficient mice.

Conclusions:

  • The developed flow cytometry approach offers a simple, efficient, accurate, and effective solution for measuring antibody affinity.
  • This method supports research in humoral immunity mechanisms, antibody affinity maturation, vaccine response evaluation, and immunoactivity/immunotoxicity testing.