Abstract
Lateral Flow Immunoassays (LFIAs) allow for rapid, low-cost, screening of many biomolecules such as food allergens. Despite being classified as rapid tests, many LFIAs take 10–20 min to complete. For a really high-speed LFIA, it is necessary to assess antibody association kinetics. By using a label-free optical technique such as Surface Plasmon Resonance (SPR), it is possible to screen crude monoclonal antibody (mAb) preparations for their association rates against a target. Herein, we describe an SPR-based method for screening and selecting crude anti-hazelnut antibodies based on their relative association rates, cross reactivity and sandwich pairing capabilities, for subsequent application in a rapid ligand binding assay. Thanks to the SPR selection process, only the fast mAb (F-50-6B12) and the slow (S-50-5H9) mAb needed purification for labelling with carbon nanoparticles to exploit high-speed LFIA prototypes. The kinetics observed in SPR were reflected in LFIA, with the test line appearing within 30 s, almost two times faster when F-50-6B12 was used, compared with S-50-5H9. Additionally, the LFIAs have demonstrated their future applicability to real life samples by detecting hazelnut in the sub-ppm range in a cookie matrix. Finally, these LFIAs not only provide a qualitative result when read visually, but also generate semi-quantitative data when exploiting freely downloadable smartphone apps.
Highlights
Lateral flow immunoassay (LFIA) is a rapid technique which relies on the fast interaction between an antibody and a target antigen [1]
As the antibodies being screened for this study were in an un-purified, crude form, a capture method was used to allow for on-chip purification and proper orientation of anti-hazelnut monoclonal antibody (mAb)
Surface plasmon resonance was used to screen antibodies in their un-purified state based on their fast association, specificity and sensitivity towards hazelnut,infor useun-purified in LFIA
Summary
Lateral flow immunoassay (LFIA) is a rapid technique which relies on the fast interaction between an antibody and a target antigen [1]. These devices have experienced a surge in popularity in the medical and food safety fields, since their birth as home-pregnancy tests [2]. It is preferred for LFIAs to use purified, fast, specific and properly characterized antibodies [3]. In order to create high-speed LFIAs, it is necessary to test the antibody rate of association towards the target analyte, as well as use a nitrocellulose membrane with a high flow rate. Traditional antibody selection techniques, such as enzyme linked immunosorbent assay (ELISA) and western blot, do not necessarily convert well into
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