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Quantification of major 2S allergen protein of yellow mustard using anti-Sin a 1 epitope antibody

Marambe, H.K., McIntosh, T.C., Cheng, B., Wanasundara, J.P.D. (2014). Quantification of major 2S allergen protein of yellow mustard using anti-Sin a 1 epitope antibody, 44 233-241. http://dx.doi.org/10.1016/j.foodcont.2014.03.053

Abstract

The major allergenic protein in yellow mustard (YM, Sinapis alba L.) is the 2S napin Sin a 1. The level of Sin a 1 in YM seeds was quantified using anti-epitope antibody (AE-Ab) generated against the epitope sequence QGPHVISRIYQTAT as the capture antibody in a non-competitive enzyme linked immunosorbent assay (NCI-ELISA), and Sin a 1 containing napin purified from YM (var Andante) as the reference standard. The AE-Ab showed high specificity towards Sin a 1 epitope-containing napin long chain and showed no cross reactivity with other proteins of YM or other Brassicaceae proteins of similar genetic homology. The assay quantified Sin a 1 with a limit of detection and quantitation (LOD and LOQ) of 3.08ppm and 3.23ppm, respectively with acceptable recoveries and precision. The Sin a 1 content in YM varieties produced in 2006, 2010, 2011 was in the range of 2.65-4.68g (AC Base); 3.81-5.98g (AC Pennant) and 3.11-4.92g (Andante) per 100g of seeds. Sin a 1 composed more than 50% of napin protein fraction of YM seed. A trend of increased Sin a 1 level with the increased contents of total seed protein and napin was observed from this data. © 2014 .

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