Abstract
We have established a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) based on two monoclonal antibodies (mAb) to measure the content of the major peanut allergen Ara h 1 in foods. Two mAbs were selected out of 12 murine hybridoma cells secreting Ara h 1-specific antibody. Using mAb 6 as the capture antibody and HRP-labelled mAb 4 as the detection antibody, the limit of detection (LOD) the assay was 0.34 ng/mL. Cross-reaction analysis showed that this method was strongly specific and had no cross-reactions with Ara h 2, pea protein or soy protein. Sample analysis showed that this ELISA was a useful tool to monitor peanut allergens in food products by measuring Ara h 1 content.
Highlights
The peanut (Arachis hypogaea L.) is a common food material and one of the most frequent causes of food allergies, accounting for approximately one-third of all severe allergic reactions [1,2]
After 15 min, the reaction was stopped by the addition of 2 M sulfuric acid (50 μL/well) and the protein concentrations were determined using a spectrophotometer at an optical density (OD) of 450 nm (OD450)
The cell supernatant from the microplate wells was tested for anti-Ara h 1 activity by indirect enzyme-linked immunosorbent assay (ELISA)
Summary
The peanut (Arachis hypogaea L.) is a common food material and one of the most frequent causes of food allergies, accounting for approximately one-third of all severe allergic reactions [1,2]. Strict avoidance of peanut-containing foods is the only possibility to prevent allergic reaction for consumers with peanut allergies [7]. Sensitization in up to 95% of peanut-allergic patients has been attributed to Ara h 1, a 65-kDa glycoprotein which comprises 12%–16% of the total protein content in peanut extracts and is an established major food allergen [9,10]. Ara h 1 presents an effective marker to monitor peanut allergen content in food products. A monoclonal antibody-based ELISA has been established to measure Ara h 1 in foods, the present study will develop a highly sensitive and more convenient sandwich ELISA [10]
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