Development of Aptamer Beacons for Antemortem Diagnosis of Chronic Wasting Disease
Annual rept. 1 May 2005-30 Apr 2006
OKLAHOMA STATE UNIV STILLWATER
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The transmissible spongiform encephalopathies TSE chronic wasting disease CWD of elk and deer has the potential for transmission to human beings. Reliable antemortem diagnostic tests for CWD are necessary for its control. Development of a novel diagnostic probe termed aptamers to detect CWD is proposed. Aptamer selections against 1 tyrosyl-tyrosyl-arginine YYR tripeptide thought to be exposed in PrPsc but not in PrPc and 2 CWD PrPP sc using a novel electrodialysis SELEX technique were assessed by 1 reduction in aptamer pool complexity by cloning and sequence analysis and 2 direct target binding assays. Neither of these selections yielded aptamers specific for PrPsc. To overcome the difficulties of removing potentially useful aptamers to CWD PrPSC during the negative selection phase, a novel antibody displaying selected motifs of PrPSC that have been shown to be involved in prion protein mis-folding will be used as our target for aptamer selection. Two such motif grafted antibodies for PrP sequences 89-112 and 136-158 were developed by Williams laboratory at the Scripps Research Institute and have graciously been provided to us through an MTA. Experiments are underway to select aptamers to the PrPSC specific motifs displayed on these antibodies. A capillary electrophoresis aptamer selection protocol is being developed for the selection process.
- Medicine and Medical Research