Purification of Nuclear RNP For Clinical and Diagnostic Application in a Autoimmune Experimental Model

VALDEZ, J.C.; LEBLANC, J.G.; AJMAT, C.; ABBAS, L.; FIAD, L.; LORENTE, C.; VALVERDE, M.; HARO, M.I.

BIOCELL

IHEM-CONICET

Lugar: Mendoza, Argentina; Año: 2006 vol. 30 p. 205 - 205

0327-9545

Mixed connective tissue disease (MCTD) is an autoimmune disorder associated with anti-ribonuclearprotein (U1-snRNP) antibodies. The aim of this study was to obtain RNP to induce an experimental model (in mice) and for use in ELISA assays. Materials and methods: Purification of RNP: Blobel and Potter´s methods was used for the collection of the nuclei of rat liver. The RNP was purified by ultracentrifugation and by fractionated precipitation. The molecular weight of the isolated protein was determined by SDS-PAGE. ELISA assays: 96 well plates were sensitized with 1 g RNP in 100 l per well. We used MCTD patients´ sera as positive controls and normal individuals´ sera as negative controls. Sera of all induced animals were tested. Results and conclusions: the molecular weight of purified RNP was compatible with one fraction of U-1 snRNP (45 kDa) and was reactive with the MCTD patients´ sera. Most animals inoculated with the RNP antigen were highly reactive, others only somewhat. Non-inoculated mice were all negative.