Method for purifying recombinant proteins expressed as...

C - Chemistry – Metallurgy – 07 – K

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C07K 1/113 (2006.01) C07K 1/36 (2006.01) C07K 14/415 (2006.01)

Patent

CA 2421438

The invention relates to a method for solubilizing and purifying recombinant proteins, which are expressed in bacterial host cells and deposited as insoluble aggregates (inclusion bodies). The purification is based on the conversion of the inclusion bodies into soluble forms while using organic denaturation reagents and on the use of chromatographic methods. To this end, inorganic, alkaline mobile solvents that contain salt are selected, which, after a purification, make it possible to provide the recombinant proteins after neutralization in a physiologically acceptable form, which can be directly employed for medical use. The method is particularly suited for purifying allergens and allergen fragments.

L'invention concerne un procédé pour solubiliser et purifier des protéines recombinées qui sont exprimées dans des cellules hôtes bactériennes et sont déposées en tant qu'agrégats non solubles ( corps d'inclusion). La purification s'effectue par transformation des corps d'inclusion en formes solides au moyen de réactifs de dénaturation organiques et par mise en oeuvre de procédés chromatographiques. A cet effet, on sélectionne des phases mobiles inorganiques, alcalines et renfermant des sels, qui permettent, une fois la purification effectuée, de mettre à disposition les protéines recombinées, après neutralisation, sous une forme physiologiquement tolérable et directement utilisable en médecine. Ce procédé est particulièrement approprié pour la purification d'allergènes et de fragments de ces derniers.

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