Multiplexed, High-Sensitivity Measurements of Antibody Affinity Using Interferometric Reflectance Imaging Sensor

Area of Science:

• Biochemistry
• Immunology
• Biomedical Engineering

Background:

• Anthrax lethal factor (LF) is a key component of anthrax toxin, driving pathogenic responses.
• Accurate kinetic measurements of ligand binding to LF are crucial for developing anthrax diagnostics and therapeutics.
• Existing methods like BLI and SPRi have limitations including signal instability and low sensitivity.

Purpose of the Study:

• To present the interferometric reflectance imaging sensor (IRIS) as a superior tool for multiplexed antibody affinity measurements against anthrax lethal factor (LF).
• To demonstrate IRIS's ability to provide kinetic information (kon and koff) and complementary binding data.
• To overcome the limitations of existing biosensing technologies for LF-targeted research.

Main Methods:

• Utilized the interferometric reflectance imaging sensor (IRIS), a label-free optical biosensor.
• Performed multiplexed antibody affinity measurements targeting anthrax lethal factor (LF).
• Compared IRIS results with traditional surface plasmon resonance (SPR) data.

Main Results:

• IRIS demonstrated high sensitivity and stability, overcoming limitations of BLI and SPRi.
• Multiplexed binding results from IRIS were consistent with SPR data.
• IRIS provided kinetic information (kon and koff) previously unattainable with SPR.

Conclusions:

• IRIS is a highly sensitive, stable, and simple biosensor for multiplexed affinity measurements.
• The IRIS system offers valuable kinetic and complementary binding data for LF research.
• IRIS advances the development of diagnostics and therapeutics for anthrax disease.