Using Mass Spectrometry to Detect Prions in Cell Lines
Produce Safety and Microbiology Research
2012 Annual Report
1a.Objectives (from AD-416):
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.
1b.Approach (from AD-416):
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. The ARS Scientists at Albany, CA have developed an extremely sensitive mass spectrometry-based approach that can be used to detect proteinase K sensitive prions. ARS Scientists at Albany, CA 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 amplify prions better than other clones. This iterative process will be used to obtain a suitable cell line and method of detection.
In FY12 the cooperator (DNA Solutions / Oklahoma State) sent samples to the ARS Scientists at Albany, CA, for analysis. Following completion of the analysis, a report was prepared and sent to the cooperator. It was determined that the immortal cell lines the cooperator had generated did not propagate infectious prions which was of interest to ARS researchers. The cooperator is currently evaluating the results and determining how to proceed. This research relates to the cell based objective of the parent project development of tests to confirm the presence or absence of transmissible spongiform encephalopathy disease agents in ingredients of animal origin and decontaminated environments.