A modular and universal platform for neutralizing-antibody profiling by quantifying antigen-receptor interactions via...

Area of Science:

• Immunology
• Molecular Biology
• Biotechnology

Background:

• Accurate detection of neutralizing antibodies (nAbs) is crucial for evaluating vaccine effectiveness and immune status against infections.
• Existing methods for nAb detection can be time-consuming or lack the sensitivity required for comprehensive immune monitoring.

Purpose of the Study:

• To develop and validate a novel, rapid, and high-throughput platform for quantifying neutralizing antibody potency.
• To demonstrate the modularity and broad applicability of the platform for detecting nAbs against different pathogens.

Main Methods:

• Developed a neutralization-antibody detection qPCR (NAD-qPCR) platform integrating antigen-receptor binding with qPCR sensitivity.
• Utilized a hybrid probe with a viral antigen module (SARS-CoV-2 RBD) and a DNA reporter, captured by magnetic beads with a receptor module (ACE2 mimic).
• Quantified nAb potency by measuring the competitive reduction in probe binding and subsequent qPCR signal.

Main Results:

• The NAD-qPCR platform demonstrated robust quantification of commercial SARS-CoV-2 nAbs with a limit of detection (LOD) of 4 ng/μL.
• Successfully profiled serum from vaccinated donors, revealing broad neutralization activity.
• Confirmed the modular design allows for easy adaptation to detect nAbs against other pathogens.

Conclusions:

• The NAD-qPCR platform provides a rapid, sensitive, and universally applicable method for nAb detection.
• This technology supports efficient vaccine development and immune monitoring by enabling high-throughput analysis of nAb responses.
• The modular design offers a flexible approach for adapting nAb detection to emerging infectious diseases.