Manufacturing Methods To Control C-Terminal Lysine, Galactose And Sialic Acid Content In Recombinant Proteins
AssigneesJanssen Biotech, Inc., Janssen Biologics B.V.
InventorsMarcel Flikweert, Charles Goochee, Francis C. Maslanka, Franciscus Johannes Ignatius Nagel, James R. Ryland, Eugene Schaefer
Life of this patent
Estimating the term…
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.
Claims29
-40. (canceled)
A method for producing an antibody sample 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 deoxyribonucleic acid (DNA) encoding an antibody in a culture medium comprising 0.74 to 2.5 μM zinc;
- and
- controlling the concentration of zinc in the culture medium, and producing the antibody sample;
- wherein the antibody is an anti-tumor necrosis factor alpha (anti-TNFα) antibody;
- and wherein
- a) the light chain of the anti-TNFα antibody comprises the complementarity determining regions (CDRs):
- CDR1, CDR2 and CDR3 sequences corresponding to amino acid residues 24-34, 50-56 and 89-97 of amino acid sequence SEQ ID NO:
- 3;
- and
- b) the heavy chain of the anti-TNFα antibody comprises the CDRs:
- CDR1, CDR2 and CDR3 corresponding to amino acid residues 31-35, 50-68 and 101-109 of amino acid sequence SEQ ID NO:
- 5;
- and
- wherein said anti-TNFα antibody is of immunoglobulin class IgG.
A method for controlling C-terminal lysine content of an antibody sample having a C-terminal lysine content of about 20% to about 70%, in a process for biosynthesizing an 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 such that the culture medium comprises 0.74 to 2.5 μM zinc, and producing the antibody sample;
- wherein the antibody is an anti-tumor necrosis factor alpha (anti-TNFα) antibody;
- and wherein
- a) the light chain of the anti-TNFα antibody comprises the complementarity determining regions (CDRs):
- CDR1, CDR2 and CDR3 sequences corresponding to amino acid residues 24-34, 50-56 and 89-97 of amino acid sequence SEQ ID NO:
- 3;
- and
- b) the heavy chain of the anti-TNFα antibody comprises the CDRs:
- CDR1, CDR2 and CDR3 corresponding to amino acid residues 31-35, 50-68 and 101-109 of amino acid sequence SEQ ID NO:
- 5;
- and
- wherein said anti-TNFα antibody is of immunoglobulin class IgG.
A method for controlling sialic acid content of an antibody sample having a sialic acid content of about 1% to about 20% in a process for biosynthesizing an 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 such that the culture medium comprises 0.74 to 2.5 μM zinc, and producing the antibody sample;
- wherein the antibody is an anti-tumor necrosis factor alpha (anti-TNFα) antibody;
- and wherein
- a) the light chain of the anti-TNFα antibody comprises the complementarity determining regions (CDRs):
- CDR1, CDR2 and CDR3 sequences corresponding to amino acid residues 24-34, 50-56 and 89-97 of amino acid sequence SEQ ID NO:
- 3;
- and
- b) the heavy chain of the anti-TNFα antibody comprises the CDRs:
- CDR1, CDR2 and CDR3 corresponding to amino acid residues 31-35, 50-68 and 101-109 of amino acid sequence SEQ ID NO:
- 5;
- and
- wherein said anti-TNFα antibody is of immunoglobulin class IgG.
A method for controlling galactose content of an antibody sample having a galactose content of about 50% to about 90% in a process for biosynthesizing an 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 such that the culture medium comprises 0.74 to 2.5 μM zinc, and producing the antibody sample;
- wherein the antibody is an anti-tumor necrosis factor alpha (anti-TNFα) antibody;
- and wherein
- a) the light chain of the anti-TNFα antibody comprises the complementarity determining regions (CDRs):
- CDR1, CDR2 and CDR3 sequences corresponding to amino acid residues 24-34, 50-56 and 89-97 of amino acid sequence SEQ ID NO:
- 3;
- and
- b) the heavy chain of the anti-TNFα antibody comprises the CDRs:
- CDR1, CDR2 and CDR3 corresponding to amino acid residues 31-35, 50-68 and 101-109 of amino acid sequence SEQ ID NO:
- 5;
- and
- wherein said anti-TNFα antibody is of immunoglobulin class IgG.
A method for controlling the ratio of sialic acid to galactose in an antibody sample 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 an antibody;
- and
- regulating the level of zinc in the culture medium during biosynthesis of the antibody such that the culture medium comprises 0.74 to 2.5 μM zinc, and producing the antibody sample;
- wherein the antibody is an anti-tumor necrosis factor alpha (anti-TNFα) antibody;
- and wherein
- a) the light chain of the anti-TNFα antibody comprises the complementarity determining regions (CDRs):
- CDR1, CDR2 and CDR3 sequences corresponding to amino acid residues 24-34, 50-56 and 89-97 of amino acid sequence SEQ ID NO:
- 3;
- and
- b) the heavy chain of the anti-TNFα antibody comprises the CDRs:
- CDR1, CDR2 and CDR3 corresponding to amino acid residues 31-35, 50-68 and 101-109 of amino acid sequence SEQ ID NO:
- 5;
- and
- wherein said anti-TNFα antibody is of immunoglobulin class IgG.
Dependencies are inferred from the claim text.