Linked Life Data

18788938

Source:http://linkedlifedata.com/resource/pubmed/id/18788938

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
2008-9-15
pubmed:abstractText
There are few reports on the isolation, quantitative recovery, and relative purification of infectious particles that cause scrapie, Creutzfeldt-Jakob disease (CJD) and epidemic bovine spongiform encephalopathy (BSE). Because pure prion protein (PrP) has failed to show significant infectivity, it is critical to find other molecules that are integral agent components. Only complex diseased tissues such as degenerating brain have been fractionated, and agent recoveries have been quite low in concentrated abnormal prion protein (PrP-res) preparations. To simplify the purification of infectious particles, we evaluated a monotypic cell line that continuously produced high levels of the 22L scrapie agent (N2a-22L). A new rapid and accurate GT1 culture assay was used to titrate infectivity in six representative sucrose gradients. We developed a streamlined approximately 3-h procedure that yielded full recovery of starting infectivity in fractions with only a few selected protein bands (representing <1% of starting protein). Infectious particles reproducibly sedimented through >30% sucrose steps, whereas PrP and PrP-res sedimentation varied depending on the conditions used. Both normal and abnormal PrP could be largely separated from infectivity in a single short centrifugation. Because no foreign enzymes were added to achieve reasonably purified infectious particles, these preparations may be used to elicit diagnostic antibodies to foreign agent proteins.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
1557-8976
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
21
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
293-302
pubmed:dateRevised
2010-10-21
pubmed:meshHeading
pubmed:year
2008
pubmed:articleTitle
Quantitative recovery of scrapie agent with minimal protein from highly infectious cultures.
pubmed:affiliation
Section of Neuropathology, Yale Medical School, New Haven, Connecticut 06510, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't, Research Support, N.I.H., Extramural