44G-9 |
The production of polyclonal antibodies against sesame seed proteins |
G. SOYLEMEZ and S. L. Hefle. Food Science and Technology, Food Allergy Research and Resource Program, University of Nebraska, Lincoln, NE 68583-0955 Sesame seed is an important allergen and can cause severe, life-threatening allergic reactions in sensitive individuals. Rapid, sensitive methods are needed to detect undeclared allergenic residues such as sesame seed, for use by the food industry to detect inadvertent contamination and assess cleaning processes. Some of the most versatile, specific, and sensitive methods for these purposes are enzyme-linked immunosorbent assays (ELISAs). The aim of the study was production of high-titer, sesame seed-specific polyclonal antibodies for the development of a sandwich ELISA for detecting potentially hazardous sesame residues in food. Polyclonal antibodies were produced at Covance Research Products, Inc.(Denver, PA). Goat, sheep and New Zealand white rabbits were immunized with defatted sesame seed powder using a standard immunization protocol. The immunogen consisted of one part defatted black (India Black) and two parts defatted white (Mexico White Hulled) sesame powder. Polyclonal antibodies were partially purified from animal serum using ammonium sulfate precipitation. Antibody production from each animal was routinely monitored using a non-competitive ELISA. Titers were defined as the log reciprocal of the midlinear portion of the resulting titration curves. The sheep and goats generated good immune responses to the sesame seed immunogen, with high titers (>10,000) and their response continued during the bleed schedule (16 months). However, rabbits gave a weak immune response to the sesame seed immunogen, with low titers (<10,000). The immunogenicity of sesame seed proteins varies between goats and sheep, and rabbits. The production of assorted polyclonal antibodies against sesame seed proteins is necessary for the development of a sandwich ELISA. An ELISA method for sesame seed would be a useful technique for the food industry to utilize to detect undeclared sesame residues in food products and on equipment, for the protection of sesame-allergic consumers.
Session 44G, Toxicology & Safety Evaluation
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