Method for purifying a gene-expression product produced by recombinant DNA technique
AssigneeThe Research Foundation for Microbial Diseases of Osaka University
InventorsHideo Goda, Toshiyuki Akiyama, Akihisa Takamizawa, Iwao Yoshida, Takeo Konobe, Keisuke Takaku
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Abstract
There is disclosed a method for purifying a gene-expression product produced by recombinant DNA technique which comprises a specific sequence of steps including adsorption treatment with silica gel, adsorption treatment with activated carbon, at least twice density gradient centrifugation and at least twice equilibrium density gradient centrifugation. The method of the present invention is very effective to remove allergen from gene-expression products contaminated therewith, enabling highly purified gene-expression products to be produced on a large scale.
Claims5
A method for purifying a gene-expression product produced by recombinant DNA technique which consists essentially of the steps of:
- (a) subjecting a crude aqueous solution containing a gene-expression product which solution is obtained from a culture of a transformant prepared by recombinant DNA technique to adsorption treatment with silica gel, thereby causing the gene-expression product to be adsorbed on said silica gel,
- (b) eluting said gene-expression product adsorbed on said silica gel to obtain an eluate containing the gene-expression product,
- (c) subjecting said eluate to adsorption treatment with active carbon, thereby causing impurities in said eluate to be adsorbed on said activated carbon, followed by recovery of the resulting eluate,
- (d) subjecting said resulting eluate treated in step (c) to density gradient centrifugation at least two times to obtain a preliminary fraction containing the gene-expression product, and
- (e) subjecting said preliminary fraction obtained in step (d) to equilibrium density gradient centrifugation at least two times to obtain a final fraction containing the gene-expression product;
- said gene-expression product being free of allergens detectable by a passive cutaneous anaphylaxis reaction test.
Dependencies are inferred from the claim text.