Manufacturing methods to control c-terminal lysine, galactose and sialic acid content in recombinant proteins
AssigneesJanssen Biologics B.V., Janssen Biotech, Inc.
InventorsMarcel Flikweert, Charles Goochee, Francis Maslanka, Franciscus Johannes Ignatius Nagel, James Ryland, Eugene Schaefer
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Abstract
Provided herein is a method for producing an antibody, such as an anti-TNFα antibody (e.g., infliximab) having a C-terminal lysine content of about 20% to about 70%, and a sialic acid content of about 1% to about 20%, comprising culturing a zinc-responsive host cell transfected with DNA encoding the antibody in a culture medium comprising at least 0.5 μM zinc; and controlling the concentration of zinc in the culture medium, thereby producing the antibody.
Claims40
A method for producing an antibody having a C-terminal lysine content of about 20% to about 70%, and a sialic acid content of about 1% to about 20%, the method comprising:
- culturing a zinc-responsive host cell transfected with DNA encoding the antibody in a culture medium comprising at least 0.5 μM zinc;
- and
- controlling the concentration of zinc in the culture medium, thereby producing the antibody.
A method for controlling C-terminal lysine content of an antibody having a C-terminal lysine content of about 20% to about 70%, in a process for biosynthesizing the antibody in a culture medium, the method comprising:
- monitoring a level of zinc in the culture medium during biosynthesis of the antibody;
- and
- regulating the level of zinc in the culture medium during biosynthesis of the antibody.
A method for controlling sialic acid content of an antibody having a sialic acid content of about 1% to about 20% in a process for biosynthesizing the antibody in a culture medium, the method comprising:
- monitoring a level of zinc in the culture medium during biosynthesis of the antibody;
- and
- regulating the level of zinc in the culture medium during biosynthesis of the antibody.
A method for controlling galactose content of an antibody having a galactose content of about 50% to about 90% in a process for biosynthesizing the antibody in a culture medium, the method comprising:
- monitoring a level of zinc in the culture medium during biosynthesis of the antibody;
- and
- regulating the level of zinc in the culture medium during biosynthesis of the antibody.
A method for controlling the ratio of sialic acid to galactose in an antibody having a ratio of sialic acid to galactose of about 0.05 to about 0.20 in a process for biosynthesizing the antibody in a culture medium, the method comprising:
- monitoring a level of zinc in the culture medium during biosynthesis of the antibody;
- and
- regulating the level of zinc in the culture medium during biosynthesis of the antibody.
Dependencies are inferred from the claim text.