|VANT-HULL, BARRY - Consultant
|DE CASTRO, ANA CECILIA RIB - Embrapa
|CHIAL, HEIDI - Biomed Bridge
|BREN-MATTISON, YVETTE - Louisiana State University
|DE BRITO, EDY - Embrapa
Submitted to: Antibodies
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/29/2021
Publication Date: 11/28/2021
Citation: Mattison, C.P., Vant-Hull, B., de Castro, A., Chial, H.J., Bren-Mattison, Y., Bechtel, P.J., de Brito, E.S. 2021. Characterization of anti-ana o 3 monoclonal antibodies and their application in comparing brazilian cashew cultivars. Antibodies. 10(4):46. https://doi.org/10.3390/antib10040046.
Interpretive Summary: Tree nut and peanut allergies are associated with significant financial, emotional, and social burdens. Allergy to cashew nuts often results in severe reactions, and there is on-going research to prevent accidental exposures and severe allergic reactions to food containing cashew nuts. Four antibodies to a potent cashew allergen called 'Ana o 3' were developed and characterized. Each of the antibodies has unique properties that can be exploited to enable new laboratory experiments to characterize Ana o 3, improve cashew detection assays, and cashew allergy therapies. For the first time, the antibodies developed here were used to screen nut extracts from eleven Brazilian cashew cultivars for differences in Ana o 3 content. Significant differences in Ana o 3 content among the cultivars were observed using the antibodies. These results may be used to guide cashew tree breeding that in the future could result in cashew nuts with decreased allergen content.
Technical Abstract: Ana o 3 is an immuno-dominant cashew nut allergen. Four monoclonal antibodies to Ana o 3 (2H5, 6B9C1, 19C9A2, and 5B7F8) were characterized by ELISA and in silico modeling. The 2H5 antibody was the only antibody specific for cashew nut extract. In addition to cashew nut extract, the 6B9C1 and 19C9A2 antibodies recognized pistachio extract, and the 5B7F8 recognized pecan extract. All four antibodies recognized both recombinant and native Ana o 3. ELISA assays following treatment of purified Ana o 3 with a reducing agent indicate the 6B9C1 and 19C9A2 antibodies likely recognize conformational epitopes, while the 2H5 and 5B7F8 antibodies likely recognize linear epitopes. In silico modeling predicted distinct epitopes for each of the anti-Ana o 3 antibodies. Screening extracts from eleven Brazilian cashew nut cultivars using all four antibodies showed slight differences in Ana o 3 binding, demonstrating that these antibodies could identify cultivars with reduced allergen content.