Process for preparing purified dimer of bone-derived factor

Details Process for preparing purified dimer of bone-derived factor Process for preparing purified dimer of bone-derived factor

C - Chemistry, Metallurgy

12

P

C12P 21/02 (2006.01) C07K 14/51 (2006.01)

Patent

CA 2275464

A process for preparing a purified dimer of a bone-derived factor, which comprises subjecting an inclusion body of a bone-derived factor produced by genetic engineering to the following steps a) to e) in sequence: a) the step of treating an inclusion body of a bone-derived factor with an unfolding agent to prepare a solubilized monomer; b) the step of treating the solubilized monomer with a refolding solution to prepare a dimer; c) the step of subjecting the refolded dimer to ultrafiltration and solvent replacement; d) the step of subjecting the dimer solution prepared above to isoelectric precipitation; and e) the step of subjecting the isoelectrically precipitated dimer to reversed phase chromatography.

L'invention a trait à un procédé permettant de préparer un dimère purifié à partir de facteur dérivé de l'os, lequel procédé consiste à faire passer un corps d'inclusion d'un facteur dérivé de l'os produit par génie génétique par les phases (a) à (e), décrites ci-dessous et ce, successivement: (a), traitement dudit corps d'inclusion au moyen d'un agent de dépliage en vue de la préparation d'un monomère solubilisé, (b), traitement du monomère solubilisé au moyen d'une solution de repliage en vue de l'obtention d'un dimère, (c), ultrafiltration du dimère replié et substitution de solvant, (d), soumission de la solution à base de dimère ainsi obtenue à une précipitation isoélectrique et (e), passage du dimère précipité par voie isoélectrique par une chromatographie en phase inverse.

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