A bioinformatics approach to the development of immunoassays for specified risk material in canned meat products |
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Authors: | Paul Reece Monique Bremer Robert Stones Christopher Danks Sabine Baumgartner Victoria Tomkies Claudia Hemetsberger Nathalie Smits Walter Lubbe |
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Institution: | (1) Food and Environment Research Agency, York, YO41 1LZ, UK;(2) RIKILT-Institute of Food Safety, Wageningen, The Netherlands;(3) Foresite Diagnostics, Sand Hutton, UK;(4) IFA Tulln, Tulln, Austria;(5) R-Biopharm AG, Darmstadt, Germany |
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Abstract: | A bioinformatics approach to developing antibodies to specific proteins has been evaluated for the production of antibodies
to heat-processed specified risk tissues from ruminants (brain and eye tissue). The approach involved the identification of
proteins specific to ruminant tissues by interrogation of the annotation fields within the Swissprot database. These protein
sequences were then interrogated for peptide sequences that were unique to the protein. Peptides were selected that met these
criteria as close as possible and that were also theoretically resistant to either pepsin or trypsin. The selected peptides
were synthesised and used as immunogens to raise monoclonal antibodies. Antibodies specific for the synthetic peptides were
raised to half of the selected peptides. These antibodies have each been incorporated into a competitive enzyme-linked immunosorbent
assay (ELISA) and shown to be able to detect the heat-processed parent protein after digestion with either pepsin or trypsin.
One antibody, specific for alpha crystallin peptide (from bovine eye tissue), was able to detect the peptide in canned meat
products spiked with 10% eye tissue. These results, although preliminary in nature, show that bioinformatics in conjunction
with enzyme digestion can be used to develop ELISA for proteins in high-temperature processed foods and demonstrate that the
approach is worth further study. |
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Keywords: | Specified risk material Bioinformatics Immunoassay Heat-processed proteins Antibody |
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