A novel approach to produce phage single domain antibody fragments for the detection of gluten in foods. (15th August 2020)

• Record Type: Journal Article
• Authors: García-García, Aina; Madrid, Raquel; González, Isabel; García, Teresa; Martín, Rosario
• Is Part Of: Food chemistry. Volume 321(2020)
• Publication Date: 2020-08-15
• Subjects: Gluten -- Celiac disease -- Phage display -- Single-domain antibody -- Antibody discovery -- ELISA -- Allergen labelling; Food -- Analysis -- Periodicals; Food -- Composition -- Periodicals; 664
• DOI: 10.1016/j.foodchem.2020.126685
• Languages: English
• ISSNs: 0308-8146

Abstract

Highlights: Phage display technology enabled the selection of gluten-affinity dAb-phage clones. The proposed methodology to produce specific dAbs avoided the use of live animals. The LOD of the indirect phage-ELISA was set at 0.01 µg/mL of PWG-gliadin. Applicability of the method was assessed through the analysis of 50 food products. Abstract: In this study, we demonstrated the feasibility of isolating recombinant phage-antibodies against gluten from a non-immunized library of human single-domain antibodies (dAbs). Phage display technology enabled the selection of affinity probes by successive rounds of biopanning against a biotinylated synthetic peptide comprising repetitive immunogenic gluten motifs. The analysis of a wide representation of heterologous plant species corroborated that two of the isolated clones were specific to wheat, barley and rye proteins. The phage antibody selected as the most appropriate clone for the detection of gluten in foods (dAb8E-phage) was further applied in an indirect ELISA to the analysis of 50 commercial food samples. Although the limit of detection achieved did not improve those of current immunoassays, the proposed methodology could provide promising new pathways for the generation of recombinant antibodies that allow a comprehensive determination of gluten in foods, whilst replacing the need for animal immunization.