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Production of pure protein antibodies and development of immunoassays to detect Ara h 3 levels in peanut varieties

Hurlburt, Barry K., Schmitt, David, Isleib, Thomas G., Cheng, Hsiaopo, Garvey, Cathryn, Koenig, Robbin L., Maleki, Soheila J.
International journal of food science & technology 2011 v.460 pp. 1477
Arachis hypogaea, allergenicity, antibodies, cultivars, epitopes, food allergies, foods, immunoassays, peanuts, protein folding, proteins, trypsin inhibitors
Peanuts are one of the most allergenic foods and are widespread in western food products. Therefore, there has been intense research into the allergic nature of the proteins involved. Ara h 3 is one of three immunodominant allergenic proteins. It is a 60-kDa protein, which forms following cleavage of the preprotein and association of the resultant 40- and 20-kDa subunits. The large subunit has been shown to harbour most of the reactive epitopes and has the protein fold likely responsible for its trypsin inhibitor activity. In this work, we have developed a method for the high-level expression and purification of recombinant Ara h 3 40-kDa subunit. Specific antibodies have been produced and applied to the secondary and tertiary screens of hundreds of peanut cultivars. Several of these cultivars were identified that have significantly reduced accumulation of Ara h 3.