| abstract |
A method of isolation and purification of a target protein by chromatography, in which chromatography eliminates or decreases the amount of prions (PrPSC), which comprises the steps of: - contacting a potentially contaminated sample with PrPSC comprising a protein target, with a multimodal chromatographic material, in which the multimodal chromatographic material contains a negatively charged 2- (benzoylamino) butanoic acid ligand; - establish the conditions of the buffer so that the target protein binds to the multimodal chromatographic material, while the PrPSCs do not bind the multimodal chromatographic material, in which the chromatographic conditions comprise the following steps: i) use a loading buffer and equilibrium containing tri-n-butyl phosphate and / or Triton X-100 in a concentration ranging from 0.1 to 10% (w / w); ii) use a wash buffer without solvent and / or non-ionic detergent; iii) use a second wash buffer containing ethylene glycol and / or lysine / arginine ranging from 5 to 30% (w / w) ethylene glycol and 0.2 to 1.5 M lysine / arginine; iv) use a third wash buffer containing sodium chloride in a concentration ranging from 0.5 to 4 M, in particular from 0.5 to 1.5 M; - followed by elution of the target protein. |