C - Chemistry – Metallurgy – 07 – K
Patent
C - Chemistry, Metallurgy
07
K
C07K 14/47 (2006.01) G01N 33/68 (2006.01)
Patent
CA 2319687
The invention concerns a method for purifying PrPres from a biological sample to be used for qualitative and/or quantitative determination of the PrPres in said sample. The method essentially consists in: (1) incubating, during 30 seconds to 2 hours, at a temperature less than 80 ·C, said biological sample with a buffer solution A comprising at least a surfactant in an amount ranging between a quarter and four times the weight of the biological sample and optionally a protease, to form a suspension S1; (2) adding to said suspension S1 resulting from (1) a buffer solution B in an amount sufficient for thinning said suspension, which buffer solution B consists of a solvent or mixture of solvents, which does not solubilize the PrPres and has a constant dielectric ranging between 10 and 25; (3) centrifuging the suspension S2 resulting from step (2); and (4) solubilizing said pellet in a buffer solution C comprising at least a surfactant and/or at least a chaotropic agent, at a temperature ranging between room temperature and 100 ·C.
Procédé de purification de la PrPres à partir d'un échantillon biologique en vue de son utilisation pour la détection qualitative et/ou quantitative de la PrPres dans ledit échantillon. Ledit procédé comprend essentiellement: (1) l'incubation, pendant 30 secondes à 2 heures, à une température inférieure à 80 ~C, dudit échantillon biologique avec un tampon A comprenant au moins un agent tensioactif en quantité comprise entre le quart et quatre fois le poids de l'échantillon biologique et éventuellement une protéase, pour former une suspension S1; (2) l'addition, à ladite suspension S1 obtenue en (1) d'un tampon B en quantité apte à éclaircir ladite suspension, lequel tampon B est constitué d'un solvant ou d'un mélange de solvants, qui ne solubilise pas la PrPres et présente une constante diélectrique comprise entre 10 et 25; (3) la centrifugation de la suspension S2 obtenue à l'étape (2); et (4) la solubilisation dudit culot dans un tampon C comprenant au moins un agent tensioactif et/ou au moins un agent chaotrope, à une température comprise entre la température ambiante et 100 ~C.
Commissariat A. L'energie Atomique
Robic
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