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United States Department of Agriculture

Agricultural Research Service

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Research Project: Using Mass Spectrometry to Detect Prions in Cell Lines

Location: Produce Safety and Microbiology Research

Project Number: 5325-32000-009-02
Project Type: Nonfunded Cooperative Agreement

Start Date: May 01, 2010
End Date: Apr 30, 2013

Objective:
The objective of this project is to develop a sensitive method of detecting the presence of prions in sample and apply this method to the development of a commercial cell-based means of detecting prions in deer and elk.

Approach:
DNA Solutions has, in collaboration with the Oklahoma State University, developed immortal deer cell lines that are capable of amplifying prions. These cells lines are among the few known to amplify CWD prions. Unfortunately, these cell lines may not produce prions that are readily detectable by Western blot. This phenomenon has been observed in other non-deer cell lines and is due to the production of proteinase K sensitive prions. Agricultural Research Service (ARS) has developed an extremely sensitive mass spectrometry-based approach that can be used to detect proteinase K sensitive prions. ARS will use this approach to quantitate the prions amplified by these cell lines. DNA Solutions or Oklahoma State may decide to sub clone these lines. The ARS approach can be used to determine if one of these clones amplifies prions better than other clones. This iterative process will be used to obtain a suitable cell line and method of detection.

Last Modified: 8/2/2014
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