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Sensitive, preclinical detection of prions in brain by nanospray liquid chromatography/tandem mass spectrometry


  • This article is a U.S. Government work and is in the public domain in the U.S.A.


More sensitive detection of prions in brain is important because it would allow early detection of disease in young animals and assure a safer food supply. We have quantitated the amount of proteinase K-resistant prion protein (PrP 27–30) by use of nano-scale liquid chromatography coupled to tandem mass spectrometry using the multiple reaction monitoring mode of operation. We used a method based on the detection of VVEQMCTTQYQK (residues 209–220) obtained by reduction, alkylation and digestion with trypsin. Quantitation of the amount of PrP 27–30 in the brains of Syrian hamsters was possible as early as 24 h after inoculation. Our results show sensitive detection of 180 fmol of PrP 27–30 per g brain (wet weight) as early as 24 h after inoculation. Clinical symptoms are not observed until 9 weeks after inoculation. Published in 2007 by John Wiley & Sons, Ltd.

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