Diagnosis of transmissible spongiform encephalopathies PrPSc/PrPC-specific antibodies

• Unité de recherche: PCRD EU
• Numéro de projet: 97.0440
• Titre du projet anglais: Diagnosis of transmissible spongiform encephalopathies PrPSc/PrPC-specific antibodies

Mots-clé (Anglais)

Prion; diagnostics; BSE; spongiform encephalopathy; antibody; immunological methods

Autre Numéro de projet (Anglais)

EU project number: BIO4CT986046

Programme de recherche (Anglais)

EU-programme: 4. Frame Research Programme - 4.1 Biotechnology

Description succincte (Anglais)

See abstract

Partenaires et organisations internationales (Anglais)

Coordinator: Universität Göttingen (D)

Résumé des résultats (Abstract) (Anglais)

The project aims at the development of a diagnostic test for live animals and humans using various antibodies. These antibodies fall into two categories: (1)Antibodies recognizing only the pathological, disease-specific form of the prion protein (PrPSc) and (2) antibodies recognizing both forms of the prion protein. The latter have been primary used to develop different post mortem detection systems for BSE such as a Western blot, an ELISA and a lateral flow chromatographic system.
In contrast to the post mortem assays, we attempt to use the prion-specific antibody 15B3 to immunoselect prions from body fluids. Since the 15B3 is an IgM antibody, we have fragmented the prion specific monoclonal antibody 15B3 to isolate the binding site in a smaller molecule. Splitting the molecule with trypsin or papain has now allowed us to have smaller fragments of the antibody which can now be coupled to various supports for direct selection of the pathological prion protein. Implementing this technology into a lateral flow format will further simplify the diagnosis of prion diseases.
An important step in the immunoaffinity purification of prions is the search for the appropriate conditions under which prions bind while the normal form of the prion protein is washed off. We have gone through a large number of detergents at different concentrations and have optimized the binding of PrPSc to 15B3. This allows us now to immunoselect prions from very dilute samples. We are now in the process of applying these condtions to various body fluids such as urine and blood. We have obtained such body fluids from normal or infected animals which in itself was a difficult task. Applying the previously identified optimal conditions will allow us now to answer the crucial question whether PrPSc is detectable in body fluids

Références bases de données (Anglais)

Swiss Database: Euro-DB of the
State Secretariat for Education and Research
Hallwylstrasse 4
CH-3003 Berne, Switzerland
Tel. [phone]
Swiss Project-Number: 97.0440