Development of sandwich ELISA for quantification of the orange allergen profilin (Cit s 2)

Abstract

Oranges (Citrus sinensis) can provoke an allergic reaction in some individuals. The concentration of profilin (Cit s 2)the major allergen in orangesis unknown, but this knowledge could be useful for preventing the development of allergies. The aim of this study was to quantify the amount of Cit s 2 in Navel oranges and in different varieties of citrus fruit. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed using both monoclonal and polyclonal antibodies. Extracts of Navel oranges and 12 citrus fruits were tested using this ELISA. Cit s 2 levels in the Navel orange pulp and peel were 1.81 ± 0.74 and 1.19 ± 0.87 µg/g, respectively, with accumulation detected primarily in the pulp. These findings may help in the production of low-risk processed oranges for orange allergy sufferers through selection of orange varieties and parts with low profilin content during food processing.

Evaluation of cleaning methods for residual orange extract on different cookware materials using ELISA with profilin allergen indicator

“…Since fresh juices are produced from various vegetables and fruits, an effective analysis method is necessary to confirm the presence or absence of residues. Profilins are actin‐binding protein components of cells (Radauer & Breiteneder, ) and are present in both the pulp and peel (Kiyota et al, ). Thus, they can be good indicators of the residue for the purpose of the study, compared with other representative allergens such as lipid transfer proteins, which are highly accumulated and detected in the peel (Iorio et al, ; Wu et al, ).…”

“…Since fresh juices are produced from various vegetables and fruits, an effective analysis method is necessary to confirm the presence or absence of residues. Profilins are actin-binding protein components of cells (Radauer & Breiteneder, 2007) and are present in both the pulp and peel (Kiyota et al, 2016).…”

“…Since profilins of birch pollen, fruits, and vegetables induce cross-allergic reactions, they are called pan-allergens (McKenna et al, 2016;Song, Zhang, Liu, & Ran, 2007). Orange profilin, which is called Cit s 2 (Iorio et al, 2013;Lopez-Torrejon et al, 2005), is considered a major allergen and present in both the pulp and peel (Kiyota et al, 2016). Additionally, other orange allergens, namely Cit s 1 and Cit s 3, accumulate mainly in the peel (Wu, Chen, Lin, Ma, & Deng, 2016).…”

Detection of the tomato allergen Sola l 1 and evaluation of its reactivity after heat and papain treatment

“…The amount of Sola l 1 in fresh tomatoes ranged from 18.0 to 40.4 µg/g, with a mean concentration of 25.6 ± 9.1 µg/g; this value was estimated to be more than 10 times higher than that in fresh oranges (Kiyota et al, 2016). In tomato-containing processed foods, such as juices and purees, Sola l 1 levels ranged from 13.3 to 24.8 µg/g.…”

“…Thus, the results of this study were obtained using rBet v 2 as a calibrator, as has been used in previous reports (Abedini et al, 2011;Ahrazem et al, 2007;Alvarado et al, 2014;Kiyota et al, 2016). In order to accurately measure the Sola l 1 concentration in the samples, corresponding recombinant or purified Sola l 1 should be prepared as standards of the calibration curve.…”