Concentration of prion protein from biological samples to increase the limits of detection by immunoassay.
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 PrPC (cellular PrP,) and PrPres (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 PrPres 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.
Davidowitz
Eliot
E
Q-RNA, Inc., 3960 Broadway, New York, NY 10032, USA. edavidowitz@q-rna.com
Eljuga
Lucy
L
Dover
Katarzyna
K
Tian
Jean
J
Grossman
Abraham
A
D000818:Animals; D000829:Animals, Domestic; D000835:Animals, Wild; D001923:Brain Chemistry; D002417:Cattle; D003670:Deer; D005260:Female; D007118:Immunoassay; D015151:Immunoblotting; D018620:PrPSc Proteins / Q000032:analysis* / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism; D034561:Sheep, Domestic
D018620:PrPSc Proteins; D011494:Protein Kinases