Method for purifying recombinant proteins expressed as...

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

Reexamination Certificate

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C530S417000, C530S419000

Reexamination Certificate

active

06900034

ABSTRACT:
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.

REFERENCES:
patent: 2002/0018779 (2002-02-01), Valenta et al.
patent: 0432419 (1991-06-01), None
patent: WO 9814467 (1998-04-01), None
Ferreira, F. et al. “Purification and Characterization of Recombinant Bel v 1, the Major Birch Pollen Allergen.” Journal of Biological Chemistry, vol. 268, No. 26, pp. 19574-19580 (1993).*
Fischer, B. “Renaturation of Recombinant Proteins Produced As Inclusion Bodies.” Biotech Adv. vol. 12, pp. 89-101 (1994).*
Lorenz A R et al., “Recombinant food allergens,” Journal of Chromatography, May 25, 2001, pp. 255-279, Amsterdam.

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