Mitigating the Hook Effect in Lateral Flow Sandwich Immunoassays Using Real-Time Reaction Kinetics

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Assay Development

Background:

• Lateral flow assays (LFAs) offer a cost-effective and user-friendly alternative to traditional laboratory diagnostics.
• Sandwich immunoassays, a common LFA format, are susceptible to the high-dose hook effect, leading to inaccurate low results at high analyte concentrations.

Purpose of the Study:

• To present a reaction kinetics-based technique to overcome the high-dose hook effect in sandwich lateral flow immunoassays.
• To significantly increase the dynamic range of LFAs for accurate analyte quantification.

Main Methods:

• Utilized a traditional sandwich lateral flow immunoassay.
• Employed a portable imaging device and a mobile interface for data acquisition and analysis.
• Applied reaction kinetics principles to analyze assay data.

Main Results:

• Successfully quantified C-reactive protein (CRP) concentrations in human serum across a wide physiological range.
• Demonstrated the technique's effectiveness in overcoming the hook effect, providing accurate results even at high concentrations.
• Achieved a significant increase in the dynamic range of the lateral flow assay.

Conclusions:

• The developed reaction kinetics technique effectively resolves the high-dose hook effect in sandwich LFAs.
• This approach offers a versatile solution applicable to various hook effect-limited sandwich LFAs.
• The technique provides high accuracy for analyte quantification, particularly within the hook effect range.