Quantification of Peach Fruit Allergen Lipid Transfer Protein by a Double Monoclonal Antibody-based Sandwich ELISA
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Quantification of Peach Fruit Allergen Lipid Transfer Protein by a Double Monoclonal Antibody-based Sandwich ELISA Zhong-shan Gao 1 & Ying-tao Ma 1,2 & Xiang Zhou 1 & Zhao-wei Yang 1 & Hui-juan Jia 1 & Ling Gao 1 & Shan-dong Wu 1 & Lu-yang Han 1 & Xue-ying Yi 1 & Hui-ying Wang 3 & Jaap H. Akkerdaas 4 & Ronald van Ree 5
Received: 14 April 2015 / Accepted: 26 July 2015 # Springer Science+Business Media New York 2015
Abstract Fruit is one of the most commonly reported food allergy sources in China, and peach lipid transfer protein (Pru p 3) has been identified as the major allergen inducing systematic symptoms. Crude allergen extracts and single component allergens have been used in food allergy diagnosis and immunotherapy. Reliable and sensitive analytical methods to quantify Pru p 3 content in fruit will help to identify lowallergenic cultivars among the abundant peach genetic resources in China. In this study, we developed a sensitive sandwich ELISA method to measure Pru p 3 in peach fruit based on two monoclonal antibodies with high sensitivity and linearity. Significant variability was observed in peach cultivars, with a much higher Pru p 3 level in peach peel than in pulp. This method will be very useful to select peach varieties with low Pru p 3 content and as a diagnostic product to detect hidden allergens in processed food. Keywords Lipid transfer protein . Monoclonal antibodies . Peach allergen . Protein quantification . Sandwich ELISA
Introduction Fruit allergy has been ranked as one of the most common food allergies in schoolchildren and adults (Gao and Gilissen 2011; Kavaliunas et al. 2012; Burney et al. 2014). In China, the number of people affected by fruit allergy has also increased with the growth in fruit consumption over the last two decades. Peach is widely planted and consumed in China, especially in the northern part, and is considered to be an important component of a healthy diet. However, it is one of the most frequently reported allergic fruit (Gao and Gilissen 2011). The majority of allergens in Rosaceae fruit have been reported to belong to four families: pathogenesis-related protein 10 (PR10 protein, birch allergen Bet v 1 homologues), thaumatin-like proteins (TLP, PR-5 proteins), nonspecific lipid transfer proteins (nsLTPs, PR-14 proteins), and profilins (PRF) (Breiteneder and Ebner 2000). In peach, these four allergen families are denoted as Pru p 1 (PR-10), Pru p 2 (TLP), Pru p 3 (LTP), and Pru p 4 (profilin) (Chen et al. 2008). Allergy to
Zhong-shan Gao and Ying-tao Ma contributed equally to this work. Electronic supplementary material The online version of this article (doi:10.1007/s12161-015-0272-x) contains supplementary material, which is available to authorized users. * Zhong-shan Gao [email protected]
3
Department of Allergy, the Second Affiliated Hospital of Zhejiang University, School of Medicine, Zhejiang 310009, People’s Republic of China
4
Department of Experimental Immunology, Academic Medical Center-University of Amsterdam, Meibergdreef 9, 1105AZ Amsterdam, T
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