18C-10

S.-H. KIM¹, T.-S. Huang², T. A. Seymour², C.-I. Wei¹, S. C. Kempf³, C. R. Bridgman³, D. Momcilovic⁴, R. A. Clemens⁵, and H. An⁵. (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) Center of Veterinary Medicine, U.S. Food and Drug Administration, Rockville, MD 20204, (5) School of Pharmacy, Univ. of Southern California, 1985 Zonal Ave., Los Angeles, CA 90089-9121

Meat and bone meal (MBM) contaminated with a transmissible spongiform encephalopathy agent was identified as the common source of bovine spongiform encephalopathy (BSE, mad cow disease) transmission. To prevent establishment and spread of BSE, the U.S. FDA introduced the feed ban, which prohibits the use of proteins derived from mammalian tissues in feeding ruminants. However, protein degradation due to the stringent rendering conditions for MBM production caused difficulties in developing adequate assay systems. The objective of this project was to develop an immunoassay for efficient detection of prohibited MBM residues in animal feed. MAb was raised against bovine smooth muscle autoclaved at 130 ºC for 20 min. The affinity of produced antibodies against autoclaved smooth muscles and MBM was tested. The enzyme-linked immunosorbent assay (ELISA) for detection of MBM in animal feed was optimized with the best performed MAb 3E10. Specificity and sensitivity of the assay was evaluated on ELISA by testing MAb 3E10 against different species of MBM and smooth muscle extracts, commercial ingredients used for animal feed, and animal feeds mixed with several different amounts of MBM. The produced MAbs showed stronger affinity against MBM than heat-treated smooth muscles at 90 ºC for 10 min. However, these antibodies showed gradual increases of reactivity as smooth muscles were further autoclaved at 130 ºC up to 1 hr. MAb 3E10 could differentiate bovine MBM from other species of MBM and ingredients used for commercial animal feeds, and detect down to 0.05% MBM mixed in animal feed. The MAbs produced against autoclaved smooth muscle extract were useful in detecting MBM. MAb 3E10 showed a great possibility to be used for effective detection of MBM in animal feed. Our study demonstrated that immunoassay could be developed as a routine analytical method for the detection of low levels of MBM in animal feed.

Session 18C, Food Chemistry: Food composition, analysis and volatiles
2:00 PM - 5:30 PM, Sunday PM Room Hall I-2

2005 IFT Annual Meeting, July 15-20 - New Orleans, Louisiana