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Concentration of prion protein from biological samples to increase the limits of detection by immunoassay
Author(s) -
Davidowitz Eliot,
Eljuga Lucy,
Dover Katarzyna,
Tian Jean,
Grossman Abraham
Publication year - 2005
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20040080
Subject(s) - immunoassay , rna , chromatography , biology , adsorption , microbiology and biotechnology , antibody , chemistry , biochemistry , immunology , gene , organic chemistry
An RNA‐ligand‐based adsorbent has been shown to concentrate prion protein (PrP) from solutions in a model system. The work presented here extends the utility of the RNA‐based adsorbent to brain homogenates of cow, sheep, mule deer ( Odocoileus hemionus ) and elk ( Cervus elaphus ). Brain homogenates were diluted either in buffer, representing specimens used in post‐mortem tests, or in serum, modelling specimens used in biological‐fluid‐based tests. The RNA adsorbent was effective in binding PrP C (cellular PrP,) and PrP res (proteinase K‐resistant PrP) from the brain homogenates of all the species tested in both model systems. The three antibodies against PrP used in the experiments identified PrP in immunoblot analysis after concentrating PrP from brain homogenates with the adsorbent, indicating the general applicability of this technology for improving the detection of PrP in immunoassays. Utilization of RNA adsorbent increased the level of detection of PrP res by immunoblot over several‐hundredfold. The results obtained suggest that this RNA adsorbent can be used to increase detection in current post‐mortem immunoassays and for the development of a blood‐based ante‐mortem test.