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Related Experiment Video

Updated: Jan 28, 2026

An Optimized Hemagglutination Inhibition HI Assay to Quantify Influenza-specific Antibody Titers

An Optimized Hemagglutination Inhibition HI Assay to Quantify Influenza-specific Antibody Titers

Published on: December 1, 2017

A digital data interpretation method for hemagglutination inhibition assay by using a plate reader.

Wei Zhao¹, Elizabeth Baldwin¹, Randall Cameron¹

¹US Horticultural Research Laboratory, 2001 South Rock Road, Fort Pierce, FL, 34945, USA.

Analytical Biochemistry

|February 25, 2019

Summary

This study introduces a digital method for reading hemagglutination inhibition (HAI) assay data, moving beyond manual observation. This digital approach offers objective and robust interpretation for glycan-lectin binding assays using chicken erythrocytes.

Keywords:

Chicken red blood cells Digital data readings Dose-response curves Hemagglutination inhibition assay IC(50)Microplate reader

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Area of Science:

Biochemistry
Assay Development
Immunology

Background:

Hemagglutination inhibition (HAI) assays quantify glycan-lectin interactions.
Current HAI data interpretation relies on subjective visual observation.
Objective and automated data analysis is needed for HAI assays.

Purpose of the Study:

To develop a digital data reading method for HAI assays.
To enable objective and robust interpretation of HAI assay results.
To automate the analysis of galectin-3-induced hemagglutination inhibition.

Main Methods:

Utilized the area scanning function of a microplate reader for digital data acquisition.
Applied the method to a galectin-3-induced hemagglutination inhibition assay.
Performed curve fitting to obtain IC50 values from OD readings.

Main Results:

Developed a digital method for HAI assay data interpretation.
Achieved a four-parameter logistic correlation (R² ≥ 0.97) between OD values and inhibitor concentrations.
Successfully obtained IC50 values, demonstrating quantitative analysis.

Conclusions:

The digital data reading method provides an objective and robust alternative to manual HAI assay interpretation.
This automated approach enhances the reliability and efficiency of glycan-lectin interaction studies.
The method is validated for HAI assays using chicken erythrocytes.