Solid phase for mixed-mode chromatographic purification of proteins
AssigneeBio-Rad Laboratories, Inc.
InventorsJiali Liao, Lee Olech, Hong Chen, Xuemei He, Russell Frost
Life of this patent
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Abstract
Proteins are purified by a mixed-mode chromatography system formed by attaching a ligand with cation exchange and hydrophobic functionalities to a large-pore support matrix, the only linkage between the ligand and the support matrix being a chain having a backbone of no more than three atoms between the hydrophobic group and the support matrix.
Claims16
A method for purifying a non-antibody protein from a source solution, said method comprising:
- (a) contacting said source solution at a pH of 4.0 to 6.0 with a mixed-mode chromatography medium comprising a ligand coupled to a solid support, said ligand comprising benzamidoacetic acid, said solid support having pores of a median diameter of 0.5 micron or greater with substantially no pores of 0.1 micron or less in diameter, and said ligand coupled to said solid support at a phenyl ring through a chain of one to three atoms, to bind said non-antibody protein in said source solution to said solid support through said ligand;
- and
- (b) eluting said non-antibody protein so bound from said solid support.
A method for purifying monomeric antibodies from a source solution comprising monomeric antibodies and antibody aggregates, said method comprising:
- (a) contacting said source solution at a pH of 4.0 to 6.0 with a mixed-mode chromatography medium comprising a ligand coupled to a solid support, said ligand comprising benzamidoacetic acid, said solid support having pores of a median diameter of 0.5 micron or greater with substantially no pores of 0.1 micron or less in diameter, and said ligand coupled to said solid support at a phenyl ring through a chain of one to three atoms, to bind said monomeric antibodies and antibody aggregates in said source solution to said solid support through said ligand;
- and
- (b) eluting said monomeric antibodies so bound from said solid support while said antibody aggregates remain bound to said solid support, thereby purifying monomeric antibodies from a source solution comprising monomeric antibodies and antibody aggregates.
A method for purifying a protein from a source solution, said method comprising:
- (a) contacting said source solution at a pH of 4.0 to 6.0 with a mixed-mode chromatography medium comprising a ligand coupled to a solid support under conditions such that the mixed-mode chromatography medium has a dynamic binding capacity of at least 40 mg human IgG per mL of mixed-mode chromatography medium, said ligand comprising benzamidoacetic acid, said solid support having pores of a median diameter of 0.5 micron or greater with substantially no pores of 0.1 micron or less in diameter, and said ligand coupled to said solid support at a phenyl ring through a chain of one to three atoms, to bind said protein in said source solution to said solid support through said ligand;
- and
- (b) eluting said protein so bound from said solid support.
Dependencies are inferred from the claim text.