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Title: MASS SPECTROMETRIC DETECTION OF ATTOMOLE AMOUNTS OF THE PRION PROTEIN, PRP 27-30, BY NANOLC-MS-MS

Author
item Onisko, Bruce
item REQUENA, JESUS - UNIV OF SANTIAGO SPAIN
item Silva, Christopher - Chris
item Dynin, Irina
item Carter, John

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 5/31/2006
Publication Date: 6/21/2006
Citation: Onisko, B.C., Requena, J., Silva, C.J., Dynin, I.A., Carter, J.M. 2006. Mass spectrometric detection of attomole amounts of the prion protein, prp 27-30, by nanolc-ms-ms. [Abstract]. 232nd ACS National Meeting and Exposition. Poster AGFD 200

Interpretive Summary:

Technical Abstract: At present there are no methods to diagnose Bovine Spongiform Encephalopathy (BSE) in live animals, or to assure a prion-free blood supply; and the result of prion infection is initiation of a neurodegenerative disease, invariably fatal after onset of symptoms. Prions have been shown to be present in blood by transfusion experiments, but based on infectivity found, the amount of misfolded prion protein in blood is estimated to be only 30-600 amol/mL. Quantitation of the prion protein was studied by use of nano-scale liquid chromatography coupled to a tandem mass spectrometer using multiple reaction monitoring. We successfully developed a method based on the detection of VVEQMCTTQYQK obtained by reduction, alkylation and digestion with trypsin of the proteinase K-resistant core known as PrP 27-30. The method exhibits a Limit of Detection of 20 amol, and has been used to quantitate the amount of PrP 27-30 in the brain of terminally ill Syrian hamsters.