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Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

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A QCM-Based Biosensor to Detect HIT-like Antibodies: Differentiating KKO from RTO via FcγRIIA Interactions.

Jianlu Zheng^1,2, Mira Diab El Harakeh³, Deokwon Seo⁴

¹Institute of Industrial Science, The University of Tokyo, Tokyo 153-8505, Japan.

Analytical Chemistry

|July 30, 2025

Summary

A new biosensor rapidly distinguishes pathogenic from nonpathogenic heparin-induced thrombocytopenia (HIT) antibodies within 10 minutes. This diagnostic tool uses FcγRIIA as a target antigen, improving HIT detection accuracy.

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Area of Science:

Biomedical Engineering
Immunology
Diagnostic Development

Background:

Heparin-induced thrombocytopenia (HIT) is a severe complication of heparin therapy.
Current diagnostic immunoassays for HIT lack specificity due to cross-reactivity with nonpathogenic antibodies.
Accurate and rapid HIT diagnosis is crucial for effective patient treatment.

Purpose of the Study:

To develop a rapid and highly selective biosensor for distinguishing pathogenic from nonpathogenic HIT antibodies.
To investigate the potential of using FcγRIIA as a diagnostic antigen for HIT.
To model HIT antibody binding using monoclonal antibodies (KKO and RTO).

Main Methods:

Utilized a quartz crystal microbalance (QCM) biosensor platform.
Employed monoclonal HIT-like antibodies (KKO) and nonpathogenic antibodies (RTO) as models.
Analyzed antibody binding affinities to FcγRIIA in the presence and absence of PF4/heparin complexes.
Measured protein zeta potentials to confirm specific binding interactions.

Main Results:

The QCM biosensor distinguished pathogenic KKO from nonpathogenic RTO antibodies within 10 minutes.
Demonstrated distinct binding affinities of KKO and RTO to FcγRIIA.
Showed that pathogenic KKO binds FcγRIIA only with PF4/heparin, while RTO binds FcγRIIA independently.
Confirmed specific binding interactions through zeta potential analysis.

Conclusions:

FcγRIIA shows potential as a superior antigen for HIT diagnostics compared to PF4/heparin complexes.
The developed biosensor offers a rapid, sensitive, and selective method for HIT antibody detection.
This approach could be extended to detect human HIT antibodies, improving diagnostic capabilities.