A bioinformatics approach to the development of immunoassays for specified risk material in canned meat products

Reece, Paul; Bremer, Monique; Stones, Robert; Danks, Christopher; Baumgartner, Sabine; Tomkies, Victoria; Hemetsberger, Claudia; Smits, Nathalie; Lubbe, Walter
August 2009
Analytical & Bioanalytical Chemistry;Aug2009, Vol. 394 Issue 7, p1845
Academic Journal
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.


Related Articles

  • Identification and Characterization of the Rat DVL2 Gene Using Bioinformatic Tools. Varişli, Lokman; Çen, Osman // Turkish Journal of Biology;2007, Vol. 31 Issue 2, p81 

    We identified and characterized the rat DVL2 gene using bioinformatics. In addition to the structure and chromosomal localization of the rat DVL2 gene, the transcribed and translated protein product of the gene was analyzed in silico. Results showed that the rat DVL2 gene consists of 15 exons...

  • Classification of protein sequences by means of irredundant patterns. Comin, Matteo; Verzotto, Davide // BMC Bioinformatics;2010 Supplement 1, Vol. 11, Special section p1 

    Background: The classification of protein sequences using string algorithms provides valuable insights for protein function prediction. Several methods, based on a variety of different patterns, have been previously proposed. Almost all string-based approaches discover patterns that are not...

  • Efficient algorithms for accurate hierarchical clustering of huge datasets: tackling the entire protein space. Yaniv Loewenstein; Elon Portugaly; Menachem Fromer; Michal Linial // Bioinformatics;Jul2008, Vol. 24 Issue 13, pi41 

    Motivation: UPGMA (average linking) is probably the most popular algorithm for hierarchical data clustering, especially in computational biology. However, UPGMA requires the entire dissimilarity matrix in memory. Due to this prohibitive requirement, UPGMA is not scalable to very large datasets....

  • DIALIGN-T: An improved algorithm for segment-based multiple sequence alignment. Subramanian, Amarendran R.; Weyer-Menkhoff, Jan; l Kaufmann, Michae; Morgenstern, Burkhard // BMC Bioinformatics;2005, Vol. 6, p66 

    Background: We present a complete re-implementation of the segment-based approach to multiple protein alignment that contains a number of improvements compared to the previous version 2.2 of DIALIGN. This previous version is superior to Needleman-Wunsch-based multi-alignment programs on locally...

  • Comparative mapping of sequence-based and structure-based protein domains. Ya Zhang; Chandonia, John-Marc; Chris6 Ding; Holbrook, Stephen R. // BMC Bioinformatics;2005, Vol. 6, p77 

    Background: Protein domains have long been an ill-defined concept in biology. They are generally described as autonomous folding units with evolutionary and functional independence. Both structure-based and sequence-based domain definitions have been widely used. But whether these types of...

  • Computational structural and functional characterization of protein family: Key for the hidden mystery. Umang, Malhotra; Astha, Jaiswal; Aastha, Chhabra; Neha, Atale; Vibha, Rani // Journal of Pharmacy Research;2012, Vol. 5 Issue 7, p3643 

    With the accretion of raw biological data in the form of unannotated protein sequences, biologists are seeking solutions to unravel the functions of proteins which have not yet been characterized experimentally. The numerous bioinformatics tools available can be used to gain an insight into...

  • NNAlign: A Web-Based Prediction Method Allowing Non-Expert End-User Discovery of Sequence Motifs in Quantitative Peptide Data. Andreatta, Massimo; Schafer-Nielsen, Claus; Lund, Ole; Buus, Søren; Nielsen, Morten // PLoS ONE;2011, Vol. 6 Issue 11, p1 

    Recent advances in high-throughput technologies have made it possible to generate both gene and protein sequence data at an unprecedented rate and scale thereby enabling entirely new ''omics''-based approaches towards the analysis of complex biological processes. However, the amount and...

  • Protein secondary structure appears to be robust under in silico evolution while protein disorder appears not to be. Schaefer, Christian; Schlessinger, Avner; Rost, Burkhard // Bioinformatics;Mar2010, Vol. 26 Issue 5, p625 

    Motivation: The mutation of amino acids often impacts protein function and structure. Mutations without negative effect sustain evolutionary pressure. We study a particular aspect of structural robustness with respect to mutations: regular protein secondary structure and natively unstructured...

  • HangOut: generating clean PSI-BLAST profiles for domains with long insertions. Kim, Bong-Hyun; Cong, Qian; Grishin, Nick V. // Bioinformatics;Jun2010, Vol. 26 Issue 12, p1564 

    Summary: Profile-based similarity search is an essential step in structure-function studies of proteins. However, inclusion of non-homologous sequence segments into a profile causes its corruption and results in false positives. Profile corruption is common in multidomain proteins, and single...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics