76D-9 |
Monoclonal antibodies against troponin I for the detection of muscle tissues after high-temperature rendering |
F. C. Chen1, Y. H. P. HSIEH1, and R. C. Bridgman2. (1) Department of Nutrition and Food Science, Auburn University, 328 Spidle Hall, Auburn, AL 36849, (2) Hybridoma Facility, Auburn University, 3410 Skyway Drive, Auburn, AL 36830
Bovine Spongiform Encephalopathy, known as "Mad Cow Disease", has had a considerable impact on the livestock industry worldwide. Regulatory controls to prevent this epidemic have prohibited the use of meat and bone meals from certain species to feed livestock. The high temperature/pressure process used in rendering animal materials presents a significant challenge in the development of analytical methods for the detection of these prohibited substances. Our objectives were (1) to produce monoclonal antibodies (MAbs) to the selected thermal-stable marker protein, troponin I (TnI), and (2) to design enzyme-linked immunosorbent assays (ELISAs) utilizing these MAbs for the detection of rendered animal muscles. Bovine skeletal TnI was used to immunize mice for the production of MAbs. The specificity of MAbs was characterized by immunoblot. Indirect ELISAs were devised to detect the presence of muscle tissues from particular species in feed samples. Six IgG class MAbs with unique specificity to TnI were selected from two consecutive fusions. The MAbs were categorized into three groups according to their reactivities to heterologous TnI. The first group contains three MAbs (1F9, 2G3, and 7F7) which display a broad reactivity to all species tested including porcine, bovine, ovine, equine, deer, chicken, turkey, duck, goose, ostrich, and catfish. The second group consists of two MAbs (7A12 and 8A12) which only react to mammalian species (porcine, bovine, ovine, equine, deer); while the third group comprises of a MAb (2A8) which recognizes specifically ruminant species (bovine, ovine, deer). Three indirect ELISAs were developed using MAbs from each groups. These assays enable the detection of heat-treated muscles (132 oC/27psi, 2h) of all animal, mammalian or ruminant species in feedstuffs at the level of 0.3, 1, and 2%, respectively. These ELISAs are valuable tools for government and animal industry to ensure the compliance with regulations and the safe production of livestock for consumer protection.
Session 76D, Muscle Foods II
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