Protein purification by Protein A chromatography
AssigneeGenentech, Inc.
InventorGregory S. Blank
Life of this patent
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Abstract
A method for purifying proteins by Protein A chromatography is described which comprises the steps of: (a) adsorbing the protein to Protein A immobilized on a solid phase comprising silica or glass; (b) removing contaminants bound to the solid phase by washing the solid phase with a hydrophobic electrolyte solvent; and (c) recovering the protein from the solid phase.
Claims17
A method for purifying a protein, which comprises a C.sub.H 2/C.sub.H 3 region, from a contaminated solution thereof by Protein A chromatography comprising:
- (a) adsorbing the protein to Protein A immobilized on a solid phase comprising silica or:
- glass;
- (b) removing contaminants bound to the solid phase by washing the solid phase with an electrolyte solvent having a pH in the range from about 5 to about 7 and comprising an electrolyte selected from the group consisting of tetramethylammonium chloride (TMAC), tetraethylammonium chloride (TEAC), tetrapropylammonium chloride and tetrabutylammonium chloride;
- and
- (c) recovering the protein from the solid.
A method for purifying a protein, which comprises a C.sub.H 2/C.sub.H 3 region, from a contaminated solution thereof by Protein A chromatography, wherein the protein has been produced by a Chinese Hamster Ovary (CHO) cell, the method comprising:
- (a) adsorbing the protein to Protein A immobilized on a solid phase comprising silica or glass;
- (b) removing Chinese Hamster Ovary Protein (CHOP) contaminants bound to the solid phase by washing the solid phase with an electrolyte solvent comprising an electrolyte selected from the group consisting of tetramethylammonium chloride (TMAC), tetraethylammonium chloride (TEAC), tetrapropylammonium chloride and tetrabutylammonium chloride;
- and
- (c) recovering the protein from the solid phase.
A method for purifying a protein, which comprises a C.sub.H 2/C.sub.H 3 region, from a contaminated solution thereof by Protein A chromatography, wherein the protein has been produced by a Chinese Hamster Ovary (CHO) cell, the method comprising:
- (a) adsorbing the protein to Protein A immobilized on a solid phase comprising silica or glass;
- (b) removing Chinese Hamster Ovary Protein (CHOP) contaminants bound to the solid phase by washing the solid phase with an electrolyte solvent having a pH in the range from about 5 to about 7 and comprising an electrolyte solvent selected from the group consisting of tetramethylammonium chloride (TMAC), tetraethylammonium chloride (TEAC), tetrapropylammonium chloride and tetrabutylammonium chloride, wherein the concentration of the electrolyte in the electrolyte solvent is in the range from about 0.25 to about 0.5 M;
- and
- (c) recovering the protein from the solid phase.
Dependencies are inferred from the claim text.