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Concentration of Disease-Associated Prion Protein with Silicon Dioxide

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Abstract

Reagents that can precipitate the disease-associated prion protein (PrPSc) are vital for the development of high sensitivity tests to detect low levels of this disease marker in biological material. Here, a range of minerals are shown to precipitate both ovine cellular prion protein (PrPC) and ovine scrapie PrPSc. The precipitation of prion protein with silicon dioxide is unaffected by PrPSc strain or host species and the method can be used to precipitate bovine BSE. This method can reliably concentrate protease-resistant ovine PrPSc (PrPres) derived from 1.69 μg of brain protein from a clinically infected animal diluted into either 50 ml of buffer or 15 ml of plasma. The introduction of a SiO2 precipitation step into the immunological detection of PrPres increased detection sensitivity by over 1,500-fold. Minerals such as SiO2 are readily available, low cost reagents with generic application to the concentration of diseases-associated prion proteins.

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Acknowledgements

The work was funded by the Department for Environment, Food and Rural Affairs; project SE2008. We thank J. Grassi (CEA Saclay, Gif/Yvette, France) for the kind gift of SAF32 and SHA31 monoclonal antibodies and the TSE-Archive at the VLA (Addlestone, Surrey, UK) for the provision of brain material. We also thank H. Simmons (VLA, Addlestone, Surrey, UK) and G. Povey (ADAS UK., Arthur Rickwood, Cambridge, UK) for the provision of ovine blood.

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Authors and Affiliations

  1. Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH, UK

    Helen C. Rees & Garry C. Whitelam

  2. ADAS UK, Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH, UK

    Ben C. Maddison & Jonathan P. Owen

  3. School of Veterinary Medicine and Science, The University of Nottingham, Sutton Bonington Campus, College Road, Sutton Bonington, Leicestershire, LE12 5RD, UK

    Kevin C. Gough

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  1. Helen C. Rees
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Correspondence to Kevin C. Gough.

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Rees, H.C., Maddison, B.C., Owen, J.P. et al. Concentration of Disease-Associated Prion Protein with Silicon Dioxide. Mol Biotechnol 41, 254–262 (2009). https://doi.org/10.1007/s12033-008-9129-5

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