Award details

Structural studies on prion proteins and related molecules

Reference	BBS/E/I/00000660
Principal Investigator / Supervisor	Dr James Hope
Co-Investigators / Co-Supervisors
Institution	The Pirbright Institute
Department	The Pirbright Institute Department
Funding type	Research
Value (£)	59,298
Status	Completed
Type	Institute Project
Start date	26/05/1997
End date	25/05/2000
Duration	36 months

Abstract

Nineteen medium-scale (~1 litre) preparations of conditioned mammalian cell culture medium containing soluble recombinant prion protein (recPrP) have been prepared, analysed and subjected to low pressure purification. Improvements in productivity have been sought throughout. The growth of cells in spinner flasks using microcarriers for support has proven to be very successful yielding > 5mg/ml recPrP. As an adjunct to the commissioning of two 7 litre bioreactor vessels, CHO cell based lines expressing recPrP are gradually being adapted to growth in low serum medium. The bioreactors and adapted cells will form the basis of the first attempts to scale up recPrP production. Along side this, gene expression constructs have been made for the expression of recPrP in myeloma cells which may ultimately be better suited to large scale production. Purified recPrP material has been shown to be correct size by mass spectrometry techniques. New stable cell lines expressing recPrP tagged with a peptide substrate for enzymatic biotinylation have been made and the product successfully biotinylated in vitro. This tagged recPrP should prove to be useful tool in both structure and function studies

Summary

unavailable

Committee	Closed Committee - Agri-food (AF)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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