67C-17 |
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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., 328 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 Bovine spongiform encephalopathy (BSE) has been a tremendous threat to both animal and human health since the first case of BSE was reported in the U.K. in 1986. It is believed that the rendered feed ingredient, meat and bone meal (MBM), contaminated with a transmissible spongiform encephalopathy agent served as the common source of infection. In 1988, the EU banned the inclusion of ruminant derived protein in animal feed. The U.S. Food and Drug Administration in 1997also banned use of MBM in animal feed to prevent the spread of BSE. Numerous efforts have been exerted to develop a method to detect MBM contaminated in animal feed. The difficulties have been encountered because of the hydrolysis of the rendered proteins in MBM resulting from high temperature and pressure treatments. In this study, a unique heat-stable biomarker was identified and used to develop an immunochemical assay system for detection of MBM residues in animal feed. The biomarker protein was purified from bovine MBM and used as an antigen to develop monoclonal antibodies. The presence of biomarker in different species of smooth and skeletal muscles, MBM, and autoclaved bovine smooth muscle was detected by SDS-PAGE and Western blot. The specific reaction of monoclonal antibodies with the biomarker was detected using Western blot and ELISA. The presence of the biomarker in MBM obtained from different manufacturers was identified by SDS-PAGE, and its molecular weight analyzed was 150 kDa. The biomarker in MBM and smooth muscle extracts was also immunostained with tested monoclonal antibodies. The antibodies could detect the biomarker in smooth muscle autoclaved at 135 degree C for up to 1 hour. The MBM biomarker identified in this study was heat-stable and suitable to raise monoclonal antibodies for the detection of MBM in animal feed.
Session 67C, Food Chemistry: Food analysis, irradiation and toxicology
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