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

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.