The susceptibility of cell lines used in the production of human biologics to prion infection

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Title: The susceptibility of cell lines used in the production of human biologics to prion infection
Authors: LeBrun, Matthew P
Date: 2009
Abstract: Transmissible spongiform encephalopathies (TSEs) are a group of neurodegenerative human and animal disorders associated with the accumulation of pathogenic or abnormal prion protein (PrPSc) in the brain. Examples include scrapie in sheep and bovine spongiform encephalopathy (BSE) in cattle. Identifying and studying pathogenic prions in vitro has been hampered by a limited capacity to detect abnormal prion proteins and incomplete knowledge of cell lines capable of sustaining a prion infection. This study focused on determining the susceptibility of cells commonly used in the manufacturing process of biologics and biotherapeutics to prion infection and on development of an enrichment procedure that used trichloroacetic acid (TCA) precipitation to increase and confirm detection of PrPSc by Western blot. The newly optimized TCA precipitation protocol demonstrated a two- to four-fold increase in sensitivity compared to other precipitating techniques when used for Western blotting. Work indicated that Sf21, N2a, RmcB, CHO, HFF, HFF-MitC, PBDC, NAMALWA, MRC-5, HEK-293, U937 and Cos-7 cell lines were not susceptible to a PrPSc infection with BSE as the inoculant strain. N2a cells were susceptible to PrPSc when a liposome-based delivery system was used. The NIH/3T3 cell line was susceptible to a PrPSc infection. This work highlighted the need to test a variety of cell lines used in the production of human biologics for their susceptibility to prion infections. Although most of the cell lines tested did not demonstrate a sustained infection, the finding that NIH/3T3 cells were susceptible indicates that cell line origin and endogenous PrPSc amounts must not be the only factors that decide if a cell lines should be screened or tested.
URL: http://hdl.handle.net/10393/28095
http://dx.doi.org/10.20381/ruor-19084
CollectionTh├Ęses, 1910 - 2010 // Theses, 1910 - 2010
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