Characterization of Anti-Ana o 3 Monoclonal Antibodies and Their Application in Comparing Brazilian Cashew Cultivars.

Christopher P Mattison,Edy Sousa De Brito,Ana Cecilia Ribeiro De Castro,Yvette Bren-Mattison,Heidi J Chial,Peter J Bechtel,Barry Vant-Hull

doi:10.3390/antib10040046

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 Ana o 3.0101 and native Ana o 3. ELISA assays following treatment of purified Ana o 3 with a reducing agent indicated that 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 11 Brazilian cashew nut cultivars using all four antibodies showed slight differences in Ana o 3 bindings, demonstrating that these antibodies could identify cultivars with varying allergen content.

Highlights

Food allergies are recognized as an increasingly important public health problem in many parts of the world
The 2H5 monoclonal antibody was generated previously using an Ana o 3.0101 peptide (H2N-CQRQFEEQQRFRNCQR−OH) immunogen and has been demonstrated to recognize properly folded and denatured native Ana o 3 purified from cashew extract, improperly folded recombinant Ana o 3.0101 produced in E. coli, and
Ana o 3, three (Ana o 3.0101) purified from E. coli, and native Ana o 3 purified from dark roasted cashew nuts

Summary

Introduction

Food allergies are recognized as an increasingly important public health problem in many parts of the world. Several factors, including culture, genetics, environment, diet, and microbiota, are thought to contribute to the increased incidence of food allergy over the past couple of decades [1]. Food allergies are associated with significant financial, emotional, and social burdens [2,3]. Several types of allergen detection tests are available, including nucleic acid amplification, mass-spectrometry, and antibody-based technologies, such as surface plasma resonance biosensors, enzyme-linked immunoassays, and point-of-care devices [4,5,6]. Cashew nuts are produced by cashew nut trees (Anacardium occidentale L.) native to Brazil. Other members of the Anacardiaceae family include pistachio, mango, poison ivy, poison oak, and sumac trees [7]

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Conclusion