Rapid Antibody Selection Using Surface Plasmon Resonance for High-Speed and Sensitive Hazelnut Lateral Flow...

Area of Science:

• Biomolecular analysis
• Assay development
• Food safety science

Background:

• Lateral Flow Immunoassays (LFIAs) are rapid, low-cost screening tools for biomolecules, including food allergens.
• Current LFIAs often require 10-20 minutes for completion, necessitating faster methods.
• Assessing antibody association kinetics is crucial for developing high-speed LFIAs.

Purpose of the Study:

• To develop and validate an SPR-based method for screening and selecting monoclonal antibodies (mAbs) for rapid LFIAs.
• To improve the speed and sensitivity of LFIAs for food allergen detection.
• To demonstrate the applicability of developed LFIAs in real-world food samples.

Main Methods:

• Utilized Surface Plasmon Resonance (SPR) for label-free screening of crude monoclonal antibody (mAb) preparations based on association rates.
• Selected anti-hazelnut mAbs based on kinetics, cross-reactivity, and sandwich pairing capabilities.
• Developed high-speed LFIA prototypes using carbon nanoparticle-labeled mAbs and assessed their performance.

Main Results:

• SPR successfully screened and selected mAbs, identifying fast (F-50-6B12) and slow (S-50-5H9) binders.
• LFIA results correlated with SPR kinetics; tests using F-50-6B12 showed a ~2x faster test line appearance (within 30 s) compared to S-50-5H9.
• Developed LFIAs detected hazelnut in a cookie matrix at sub-ppm levels and provided both qualitative and semi-quantitative results via smartphone apps.

Conclusions:

• SPR is an effective tool for selecting high-affinity antibodies to accelerate LFIA development.
• The developed high-speed LFIAs offer rapid, sensitive, and versatile detection of food allergens.
• These LFIAs show promise for real-world food safety applications, providing accessible data analysis.