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-  2019 

A cashew specific monoclonal antibody recognizing the small subunit of Ana o 3

DOI: 10.1016/j.toxrep.2019.06.018

Keywords: Food allergy, Cashew, Ana o 3, Antibody, ELISA

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Abstract:

Food allergies represent a substantial medical liability and preventing accidental exposure to food allergens requires constant attention. Allergic reaction to cashew nuts is frequently serious, and the small 2S albumin, Ana o 3, is an immuno-dominant cashew allergen. Ana o 3 is composed of five alpha helices, contains 2 subunits linked by cysteine disulfide bonds, and remains soluble even after extensive heating of cashew nuts. The stability and solubility properties of Ana o 3 make it an excellent target for diagnostic and detection methods and tools. In this work, a monoclonal antibody, designated 2H5, aimed at amino acids 39–54 within helices I and II of the small subunit of Ana o 3 was developed that recognizes both recombinant and native Ana o 3 and is cashew specific in ELISA experiments. The KD against the targeted amino-acid sequence was found to be approximately 7.0?×?10?6 mg/ml (3.3?nM), while the KD against the native protein was found to be approximately 1.2?×?10-3 mg/ml (92?nM). The 2H5 monoclonal anti-Ana o 3 antibody can distinguish between native and recombinant proteins and represents a useful reagent for the study of antibody cashew-allergen interactions and may enable the development of cashew-specific diagnostic tools that can be used to prevent accidental cashew allergen exposures

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