Integrated approach to the isolation and purification of antibodies
AssigneeAbbott Laboratories
InventorsDiane D. Dong, Stephen M. Lu, Natarajan Ramasubramanyan, Wen-Chung Lim
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Abstract
Disclosed herein is an integrated approach to purification process development and execution, including processes comprising particular capture and fine purification steps; processes that employ of a minimal number of buffer systems, and processes that make use of minimally-corrosive buffer systems, as well as combinations thereof.
Claims20
A method for producing a host cell protein-reduced antibody preparation from a sample mixture comprising an antibody and at least one host cell protein, said method comprising:
- (a) contacting said sample mixture with a loading buffer and contacting said loading buffer and sample mixture to a capture separation chromatographic support under conditions where said antibody is retained on said chromatographic support;
- (b) washing said capture separation chromatographic support with a wash buffer to remove the sample mixture components that are not retained on said capture separation chromatographic support;
- and
- (c) contacting said capture separation chromatographic support with an elution buffer to thereby produce a capture separation eluate;
- wherein said loading, wash, and elution buffers consist of water and essentially the same anion and cation components;
- and
- wherein said capture separation eluate comprises said host cell protein-reduced antibody preparation.
A method for developing an integrated purification protocol for producing host cell-reduced preparations from two sample mixtures where each sample mixture comprises a distinct antibody and at least one host cell protein:
- (a) selecting a capture separation chromatographic support capable of retaining the distinct antibodies of said sample mixtures;
- and
- (b) selecting load, wash, and elution buffers for, respectively, loading, washing, and production of a capture separation eluate;
- wherein said loading, wash, and elution buffers consist of water and essentially the same anion and cation components;
- and
- wherein said capture separation eluate comprises said host cell protein-reduced antibody preparation.
A method for producing a host cell protein-reduced antibody preparation from a sample mixture comprising an antibody and at least one host cell protein, said method comprising;
- (a) contacting said sample mixture with a loading buffer and contacting said loading buffer and sample mixture to a capture separation chromatographic support under conditions where said antibody is retained on said chromatographic support;
- (b) washing said capture separation chromatographic support with a wash buffer to remove the sample mixture components that are not retained on said capture separation chromatographic support;
- (b) contacting said capture separation chromatographic support with an elution buffer to thereby produce a capture separation eluate comprising said antibody;
- (c) contacting said capture separation eluate to a second capture separation chromatographic support under conditions where said antibody is retained on said second capture separation chromatographic support, wherein said second capture separation chromatographic support is selected from the group consisting of an ion exchange matrix, a mixed mode resin and a hydrophobic interaction resin;
- and
- (d) contacting said second capture separation chromatographic support with an elution buffer to thereby produce a second capture separation eluate,
- wherein said loading, wash, and elution buffers consist of water and essentially the same anion and cation components;
- and
- wherein said second capture separation eluate comprises said host cell protein-reduced antibody preparation.
Dependencies are inferred from the claim text.