title: Production and characterization of rabbit polyclonal antibodies to almond (Prunus dulcis L.) major storage protein.
abstract: Rabbits were immunized with purified almond major protein (AMP), the primary storage protein in almonds. Rabbit anti-AMP polyclonal antibodies (PA) could detect AMP when as little as 1-10 ng/mL were used to coat microtiter plates in a noncompetitive enzyme linked-immunosorbent assay (ELISA). Competitive inhibition ELISA assays detected the AMP down to 300 ng/mL. PA recognized the AMP in protein extracts from all U.S. major marketing cultivars of almonds (Mission, Neplus, Peerless, Carmel, and Nonpareil) with specific reactivity of 52.6-75% as compared to that of the AMP alone. Immunoreactivity of protein extracts prepared from commercial samples of blanched almonds, roasted almonds, and almond paste was respectively reduced by 50.0%, 56.6%, and 68.4% (noncompetitive ELISA) when compared to the immunoreactivity of the AMP. Moist heat (121 degrees C, 15 min) pretreatment of the AMP reduced the PA reactivity by 87% (noncompetitive ELISA). Exposing AMP to pH extremes (12.5 and 1.5-2.5) caused a 53% and 57% reduction in PA reactivity, respectively (noncompetitive ELISA). PA showed some cross-reactivity with the cashew major globulin, and to a lesser extent, the Tepary and Great Northern bean major storage protein (7S or phaseolin). The presence of almonds in a commercial food was detected using PA in a competitive ELISA.
authors.affiliation: Department of Nutrition, College of Human Sciences, Florida State University, Tallahassee, Florida 32306-1493, USA.
authors.forename: M R
authors.initials: MR
authors.lastname: Acosta
authors.forename: K H
authors.initials: KH
authors.lastname: Roux
authors.forename: S S
authors.initials: SS
authors.lastname: Teuber
authors.forename: S K
authors.initials: SK
authors.lastname: Sathe
mesh_terms: D000485:Allergens / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D000818:Animals; D000917:Antibody Formation; D004797:Enzyme-Linked Immunosorbent Assay; D005512:Food Hypersensitivity / Q000209:etiology / Q000276:immunology*; D006801:Humans; D009754:Nuts / Q000009:adverse effects / Q000737:chemistry* / Q000276:immunology; D010936:Plant Extracts / Q000009:adverse effects / Q000032:analysis / Q000276:immunology; D010940:Plant Proteins / Q000009:adverse effects / Q000032:analysis / Q000276:immunology*; D011817:Rabbits
chemical_list: D000485:Allergens; D010936:Plant Extracts; D010940:Plant Proteins