Antibody purification
AssigneeSmith Kline Beecham Corporation
InventorsPaula J. Shadle, John C. Erickson, Robert G. Scott, Thomas M. Smith
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Abstract
This invention relates to the application of hydrophobic interaction chromatography combination chromatography to the purification of antibody molecule proteins.
Claims41
A method for purifying monomeric IgG antibody from a mixture comprising said monomeric antibody and at least one of immunoglobulin aggregates, misfolded species, host cell protein or protein A comprising contacting said mixture with a hydrophobic interaction chromatographic support and selectively eluting the monomer from the support.
A method for the purification of an IgG antibody from conditioned cell culture medium containing same comprising sequentially subjecting the medium to (a) Protein A affinity chromatography, (b) ion exchange chromatography, and (c) hydrophobic interaction chromatography.
A method for purifying antibody from a conditioned cell medium comprising:
- (a) adsorbing the antibody onto a Protein A chromatographic support;
- (b) washing the adsorbed antibody with at least one buffer;
- (c) eluting the antibody from step (b);
- (d) adsorbing the antibody from step (c) onto an ion exchange chromatographic support;
- (e) washing the absorbed antibody with at least one buffer;
- (f) selectively eluting the antibody from step (e);
- (g) adsorbing the eluate of step (f) onto a hydrophobic interaction chromatographic support;
- (h) washing the adsorbed antibody with at least one buffer;
- (i) eluting the adsorbed antibody;
- and
- (j) recovering the antibody.
A method of removing Protein A from a mixture comprising Protein A and IgG antibodies comprising contacting the mixture with a hydrophobic interaction chromatography support and selectively eluting the antibody from the support.
Dependencies are inferred from the claim text.