Award details

The development of antibodies specific for conformational isomers of recombinant ovine PrP and their evaluation as diagnostics tools

Reference	BSD17727
Principal Investigator / Supervisor	Professor Garry Whitelam
Co-Investigators / Co-Supervisors	Professor Kevin Gough, Mr R Jackman
Institution	University of Leicester
Department	Biology
Funding type	Research
Value (£)	183,412
Status	Completed
Type	Research Grant
Start date	01/07/2002
End date	30/09/2005
Duration	39 months

Abstract

Currently, all TSE diagnostics are post mortem immunoassays applied to central nervous system (CNS) tissues and are able only to identify symptomatic animals, or in some cases those nearing clinical disease. The limitation of such tests is primarily one of analytical sensitivity inherent in the need for sample pretreatment involving digestion with protein kinase K or differential solubilisation in chaotropic agents to eliminate the normal cellular form of the prion protein (PrPc). These procedures significantly reduce the levels of the disease isoform (PrPsc) in the extract applied to the endpoint immunoassay and may also only target a percentage of the PrPsc present that is in an aggregated form. To extend the use of CNS as the analytical matrix in animals of earlier age and to apply immunoassays to lymphoreticular system tissues (post mortem) and body fluids (pre-mortem/live animal), destructive sample preparation systems must be eliminated by the development of immunoreagents capable of binding to PrPsc specifically in the presence of PrPc. This proposal seeks to produce such antibodies by phage display through the use of the beta-form of recombinant PrP. This beta-sheet rich conformation will be used as a pure panning target for the selection of binders from both naive phage display antibody libraries and from immune libraries created from rabbits immunised with the beta-form of PrP. Resultant soluble antibodies will be evaluated in a range of immunoassays including ELISA and immunoblots following their demonstration of specificity by immunoprecipitation of PrPsc. Appropriate immunoassays will then be applied to a range of clinical and pre-clinical tissues and body fluids from TSE infected animals.

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	Biology Spongiform Encephalopathies - Diagnostics (BSD) [2000]
Funding Scheme	X – not Funded via a specific Funding Scheme

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