Quantitative detection of allergenic protein Sin a 1 from yellow mustard (Sinapis alba L.) seeds using enzyme-linked immunosorbent assay.
Allergy to yellow mustard (YM; Sinapis alba L.) seed proteins has been reported and is currently seen as a constraint that hampers expansion of YM protein utilization. The most predominant allergenic protein of YM seed has been recognized as Sin a 1. In this study, Sin a 1 was purified ( S. alba var. Andante), rabbit polyclonal antibodies (pAb) specific to Sin a 1 were generated, and a sandwich enzyme-linked immunosorbent assay (S-ELISA) was developed to detect and quantify Sin a 1 from YM. The S-ELISA method using Sin a 1-pAb and its horseradish peroxidase conjugate resulted in a detection limit of 0.3 microg/mL for purified Sin a 1. The Sin a 1 contents of six YM lines were in the range of 0.82-2.94 mg/g when assayed by the developed S-ELISA method. The results showed that S-ELISA could distinguish Sin a 1 in YM seed-derived extracts rapidly and could be applied in controlling and/or monitoring of YM allergenic proteins.
Shim
Youn-Young
YY
Saskatoon Research Centre, Agriculture and Agri-Food Canada, Saskatchewan, Canada.
Wanasundara
Janitha P D
JP
D000485:Allergens / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D052179:Antigens, Plant; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005504:Food Analysis / Q000379:methods; D005506:Food Contamination / Q000032:analysis*; D005512:Food Hypersensitivity / Q000517:prevention & control*; D006801:Humans; D010940:Plant Proteins / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D031226:Sinapis / Q000737:chemistry* / Q000276:immunology
D000485:Allergens; D052179:Antigens, Plant; D010940:Plant Proteins; C079479:Sin a I protein, Sinapis alba