Quantification of Pla or 3, a Platanus orientalis Allergen, Grown under Different Environmental Conditions, by Sandwich ELISA

• Main Authors: Farnaz Sedghy, Mojtaba Sankian, Maliheh Moghadam, Abdol-Reza Varasteh
• Format: Article
• Language: English
• Published: Varastegan Institute for Medical Sciences 2016-10-01
• Series: Reports of Biochemistry and Molecular Biology
• Subjects: Pathogenesis-related proteins; Pla or 3; Platanus orientalis; Sandwich ELISA
• Online Access: http://rbmb.net/article-1-84-en.pdf

Background: Platanus species are widely cultured around the world and considered an important cause of allergic reactions. In the present study, we developed a sandwich ELISA to quantify Pla or 3 allergen in P. orientalis pollen extracts grown near high-traffic roads and compared it to pollen extracts collected from rural areas as control. Methods: Pollen samples were collected from three polluted and two unpolluted sites in Mashhad, northeast Iran. Recombinant Pla or 3 was expressed and used for polyclonal antibody production in rabbit. A sandwich ELISA was developed and validated to quantify Pla or 3 levels in pollen extracts from the different sites. Results: The coefficients of variation (CVs) for the intra- and inter- day assays were less than 5 and 18%, respectively. The working range of the standard curve was between 0.1 and 25 ng/ml, with the detection limit being 0.037 ng/ml. The recovery percentage was 88-106.4% at working concentrations from 0.31 to 26.5 ng/ml. Pla or 3 levels were significantly greater in pollens grown near high-traffic roads than in those grown in rural regions (p < 0.0001). Conclusions: A sandwich ELISA was developed and validated to quantify Pla or 3 in pollen extracts. Using this validated ELISA, we showed a substantial difference between the amounts of Pla or 3 in pollens grown in different environments. This finding should be considered in developing public policies to reduce traffic pollution, which leads to reduced allergic reactions in atopic subjects.