NR ATJZ
AU Cecillon,S.; Moussa,A.; Dupin,M.; Perron,H.; Coleman,A.W.
TI The Use of para-Sulphonato-calix[n]arenes to Amplify Detection of PrPsc in an ELISA assay
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Diagnosis DIA-11
PT Konferenz-Poster
AB
The calix[n]arenes are a class of synthetic macrocyclic receptors widely studied in Supramolecular chemistry for their ability to complex a wide range of neutral and charged small organic molecules and also inorganic ions. The biological properties of these molecules have been much slower to come to the fore, however since the 1990s there has been a growing recognition of their potential to interact with proteins.
In the previous communication we demonstrated the use of para-Sulphonato-calix[n]arenas in a Western Blot assay of PrPsc. However, such an assay is not conclusive to use a widely applicable diagnostic test for PrPsc, such a test requires the transposition to an ELISA format. Such a transposition required the development of a highly reproducible chemistry for coupling the para-Sulphonato-calix[n]arene molecule used in the ELISA test the 96 well titre plates with retention of the amplification effect of the para-Sulphonato-calix[n]arenes.
Here we present the methodology developed to covalent couple the para-Sulphonato-calix[n]arenes to ELISA plates having N-Hydroxysuccinimde surface activation. We will demonstrate with regard to PrP detection:
1. The excellent sensitivity of the para-Sulphonato-calix[n]arene derived ELISA assay
2. The application of the para-Sulphonato-calix[n]arene derived ELISA assay to different physiological fluids
3. The high degree of reproducibility of the chemistry used
The studies described have led to the integration of the para-Sulphonato-calix[n]arene derived ELISA assay into a ante-mortem diagnostic test for PrPsc which can be applied to the detection of PrPsc in the blood (see other communications).
AD S.Cecillon, A.W.Coleman, Institut de Biologie et de Chimie des Protéines (CNRS), Lyon, France.; A.Moussa, Agence Française pour la Sécurité Sanitaire des Aliments, Lyon, France; M.Dupin, H.Perron, BioMérieux R&D Immunoassay and Proteomics Dept., Marcy L'Etoile, France
SP englisch
PO Deutschland