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Protein
Markers for Evaluation of Prion Inactivation
Fur-Chi
Chen, Agnes Kilonzo-Nthenge, Dwyan Young, and Abdullah Abdullah
Institute of
Agricultural and Environmental Research
Seminar Series
Tennessee State University, Nashville, TN
February 16, 2005
Bovine
Spongiform Encephalopathy (BSE) is thought to enter the human food chain
through beef products contaminated with infectious prion proteins.
Developing strategies for the inactivation of BSE infectivity in
processed meat products would provide further safeguards for the
Nation’s food supply. Studies have demonstrated the potential
applications of emerging inactivation technology in meat processing.
However, lacking of a rapid verification measurement to facilitate the
evaluation of such technology has hampered its further developments. The
overall goals of this project are to identify indigenous protein markers
to serve as surrogate agents for prion proteins, and to study
denaturation of the protein markers using monoclonal antibody (MAb)
immunoassay for verifying efficacy of the inactivation procedure.
Protein markers from muscle tissues were selected based on their heat
and protease resistance. A panel of ten MAbs to tropomyosin, the protein
marker for muscle model, has been produced. These MAbs were
characterized into three groups by indirect enzyme-linked immunosorbent
assay (ELISA) and Western blotting according to their reactivity to
heat-treated tropomyosin. Epitope mapping by competitive ELISA was
performed to determine topology of the binding of MAbs to tropomyosin.
These MAbs were categorized into five distinct epitopes and the
potential combinations of the paired-antibodies to be used in sandwich
ELISA were identified. The sandwich ELISA employing these MAbs has been
constructed to quantify the extent of denaturation of tropomyosin under
various heat treatments. The results indicated that tropomyosin could be
a useful marker for monitoring the inactivation of BSE agent in meat
products.
Funding Agency: FDA-CFSAN
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