Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 104, Issue 52, Pages 20914-20919Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0710152105
Keywords
prion protein; PrP(sc); thioflavin T; protease-sensitive; femtogram
Categories
Funding
- NIA NIH HHS [P01 AG010770, P01 AG021601, AG021601, AG10770, P01 AG002132, AG02132] Funding Source: Medline
Ask authors/readers for more resources
Polymerization of recombinant prion protein (recPrP), which was produced in bacteria, into amyloid fibers was accompanied by the acquisition of prion infectivity. We report here that partially purified preparations of prions seed the polymerization of recPrP into amyloid as detected by a fluorescence shift in the dye Thioflavin T. Our amyloid seeding assay (ASA) detected PrP(sc), the sole component of the prion, in brain samples from humans with sporadic Creutzfeldt-Jakob disease, as well as in rodents with experimental prion disease. The ASA detected a variety of prion strains passaged in both mice and hamsters. The sensitivity of the ASA varied with strain type; for hamster Sc237 prions, the limit of detection was approximate to 1 fg. Some prion strains consist largely of protease-sensitive PrPsc (sPrP(sc)), and these strains were readily detected by ASA. Our studies show that the ASA provides an alternative methodology for detecting both sPrP(sc) and protease-resistant PrP(sc) that does not rely on protease digestion or immunodetection.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available