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AssigneeMerck Patent GmbH
InventorsRoland Suck, Oliver Cromwell, Helmug Fiebig
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The invention relates to a method for solubilising and purifying recombinant proteins which are expressed in bacterial host cells and are deposited as insoluble aggregates (inclusion bodies). The purification is based on conversion of the inclusion bodies into soluble forms using organic denaturing reagents and using chromatographic methods. Inorganic, alkaline, salt-containing eluents are selected here which, after purification is complete, enable the recombinant proteins, after neutralisation, to be made available in a form which can be employed directly for medical use and is physiologically acceptable. The method is particularly suitable for purifying allergens and allergen fragments.
Method for the preparation of purified, recombinant proteins in soluble, biologically active form as a physiological medium which is directly suitable for medical use from protein aggregates (“inclusion bodies”) obtained after bacterial expression which are insoluble in the expression medium,
Method according to one of claims 3 to 7, characterised in that at least one further chromatography purification step is carried out.
Method according to claim 8, characterised in that the further chromatography step carried out is hydrophobic interaction chromatography and/or gel filtration.
Dependencies are inferred from the claim text.