67C-18 |
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S.-H. KIM1, T.-S. Huang2, T. A. Seymour2, C.-I. Wei1, S. C. Kempf3, C. R. Bridgman3, R. A. Clemens4, and H. An4. (1) Dept. of Nutritional Sciences, Oklahoma State Univ., 301 HES, Stillwater, OK 74078, (2) Dept. of Nutrition & Food Science, Auburn Univ., 210 Spidle Hall, Auburn, AL 36849, (3) Hybridoma Facility, Auburn Univ., Dept. of Biological Sciences, 113 Cary Hall, Auburn, AL 36849, (4) School of Pharmacy, Univ. of Southern California, 1985 Zonal Ave., Los Angeles, CA 90089-9121 Meat and bone meal (MBM) is utilized to supplement nitrogen need in animal feed. However, MBM contaminated with a transmissible spongiform encephalopathy (TSE) agent was identified as the common source of bovine spongiform encephalopathy transmission. The strategy of the U.S. Food and Drug Administration to prevent the establishment and spread of BSE in the U.S. is based on the control of animal feed. In 1997, the FDA introduced the feed ban of MBM, which prohibits the use of proteins derived from mammalian tissues in feeding ruminants. For efficient control of the feed ban, it is necessary to develop an assay method to detect the presence of MBM in animal feed. In this study, an immunological assay was developed for the detection of MBM residues in animal feed. Antibodies were raised against a heat-stable protein purified from bovine smooth muscle. Hybridoma technique was applied to produce monoclonal antibodies. Approximately, 1,000 hybridoma cells were screened for each of the fusions, and selected cells were cloned. The specificity of the assay was evaluated on ELISA by testing the antibodies against different species of MBM and smooth muscle extracts and commercial ingredients used for animal feed. The sensitivity of the assay was determined by testing the monoclonal antibodies against animal feeds mixed with several different amounts of MBM. Obtained antibodies reacted with all the MBM extracts tested. All of the antibodies showed good interactions with bovine smooth muscle extract treated at 135 degree C for up to 1 hr. The antibodies could differentiate MBM from most of the ingredients used for animal feed. The antibodies were able to detect 0.1% of MBM mixed in animal feed. The hybridoma technique was suitable to provide MBM-specific antibodies. The tested antibodies showed the potential possibility for sensitive detection of prohibited MBM residues in animal feed.
Session 67C, Food Chemistry: Food analysis, irradiation and toxicology
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