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Immunoaffinity reactors for prion protein qualitative analysis

Authors: Bílková Zuzana | Castagna Annalisa | Zanusso Gianluigi | Farinazzo Alessia | Monaco Salvatore | Damoc Eugen | Przybylski Michael | Beneš Milan | Lenfeld Jiří | Viovy jean-Louis | Righetti Pier Giorgo

Year: 2005

Type of publication: článek v odborném periodiku

Name of source: Proteomics

Publisher name: Wiley-Blackwell

Page from-to: 639-647

Titles:

Language	Name	Abstract	Keywords
cze	Imunoafinitní reaktory v kvalitativní analýze prionového proteinu	Imunoafinitní reaktory v kvalitativní analýze prionového proteinu	mozkový homogenát; imunosorbent; magnetické mikročástice; prionový protein
eng	Immunoaffinity reactors for prion protein qualitative analysis	The cellular prion protein (PrPc) represents the substrate for generation of conformational aberrant PrP isoforms which occur in human and animal prion diseases. The published two-dimensional maps of human PrPc show a vast microheterogeneity of this glycoprotein. The main goal of this project was to develop a highly specific immunoaffinity reactor for qualitative analysis of PrP cellular isoforms isolated from brain homogenate, cerebrospinal fluid and other tissues. New techniques for affinity proteomics, carriers and immobilization chemistry were applied. The choice of matrix (chemical and magnetic properties, particle size and distribution, porosity) was the key factor that influenced the quality of the reactor and the nature of final applications. Mouse anti-prion IgGs directed to N-terminal and C-terminal epitopes (residues 23-40 and 147-165) were grafted in different manners to magnetic micro- and nanoparticles particularly developed for micro-CHIP application. High operational and storage stability of affinity reactors with minimized nonspecific absorption were achieved. The quality of the immunoreactors was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by immunoblotting and by two-dimensional electrophoresis.	brain homogenat; immunosorbent; magnetic microspheres; prion protein