Granted Patent B2

US 12,655,385 · App. 19/325,217 · Granted Jun 16, 2026

Inventors: Natarajan Vijayasankaran (South San Francisco, CA); Steven J. Meier (South San Francisco, CA); Sharat Varma (South San Francisco, CA); Yi Yang (South San Francisco, CA)

Assignee: Genentech, Inc.

C12N5/0018C07K16/00C07K2317/14C12N2500/30C12N2500/33C12N2500/44C12N2500/46

Abstract

Claims (53)

1 . A method of making an anti-CD20 antibody composition comprising:

(a) culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding the anti-CD20 antibody in a cell culture medium comprising one or more of components (i)-(v):

(i) hypotaurine,

(ii) s-carboxymethylcysteine,

(iii) butylated hydroxyanisole,

(iv) quercitrin hydrate, and

(v) aminoguanidine;

(b) producing the anti-CD20 antibody using the CHO cells;

(c) purifying the anti-CD20 antibody from the cell culture medium; and

(d) concentrating the anti-CD20 antibody of step (c) to make the anti-CD20 antibody composition.

2 . The method of claim 1 , the wherein the anti-CD20 antibody has reduced oxidation in the cell culture medium comprising one or more of components (i)-(v).

3 . The method of claim 1 , the wherein the anti-CD20 antibody composition after step (d) has reduced color intensity when the anti-CD20 antibody is produced by the CHO cells cultured in the cell culture medium.

4 . The method of claim 1 , wherein the concentration of the anti-CD20 antibody in the anti-CD20 antibody composition is at least 100 mg/mL.

5 . The method of claim 1 , wherein the concentration of the anti-CD20 antibody in the anti-CD20 antibody composition is about 150 mg/mL to about 250 mg/mL.

6 . The method of claim 1 , wherein the cell culture medium comprises hypotaurine at a concentration from about 2.0 mM to about 50.0 mM.

7 . The method of claim 1 , wherein the cell culture medium comprises s-carboxymethylcysteine at a concentration from about 8.0 mM to about 12.0 mM.

8 . The method of claim 1 , wherein the cell culture medium comprises butylated hydroxyanisole at a concentration from about 0.025 mM to about 0.040 mM.

9 . The method of claim 1 , wherein the cell culture medium comprises quercitrin hydrate at a concentration from about 0.010 mM to about 0.020 mM.

10 . The method of claim 1 , wherein the cell culture medium comprises aminoguanidine at a concentration from about 0.0003 mM to about 10 mM.

11 . The method of claim 1 , wherein the culturing comprises a fed batch cell culture process.

12 . The method of claim 1 , wherein the CHO cells are cultured using a 14-day cell culture cycle.

13 . The method of claim 1 , wherein the one or more of components (i)-(v) are included in the cell culture medium on day 0 of a 14-day cell culture cycle.

14 . The method of claim 1 , wherein the one or more of components (i)-(v) are added to the cell culture medium on one or more days of a 14-day cell culture cycle.

15 . The method of claim 1 , wherein the CHO cells are cultured within a range of 25° C. to 38° C.

16 . The method of claim 1 , wherein the CHO cells are cultured within a range of 35° C. to 40° C.

17 . The method of claim 1 , wherein the anti-CD20 antibody is an IgG1 monoclonal antibody.

18 . The method of claim 1 , wherein the anti-CD20 antibody composition is formulated for subcutaneous injection.

19 . A pharmaceutical formulation comprising the anti-CD20 antibody composition of claim 1 in a pharmaceutically acceptable carrier.

20 . The pharmaceutical formulation of claim 19 , wherein the anti-CD20 antibody composition is formulated for subcutaneous injection.

21 . The pharmaceutical formulation of claim 20 , wherein the pharmaceutical formulation is sterile.

22 . A method of making a pharmaceutical formulation comprising combining:

(1) a concentrated anti-CD20 antibody composition, and

(2) a pharmaceutically acceptable carrier;

wherein, the concentrated anti-CD20 antibody composition is made by:

(a) culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding the anti-CD20 antibody in a cell culture medium comprising one or more of components (i)-(v):

(i) hypotaurine,

(ii) s-carboxymethylcysteine,

(iii) butylated hydroxyanisole,

(iv) quercitrin hydrate, and

(v) aminoguanidine;

(b) producing the anti-CD20 antibody using the CHO cells;

(c) purifying the anti-CD20 antibody from the cell culture medium; and

(d) concentrating the anti-CD20 antibody to make the concentrated anti-CD20 antibody composition.

23 . The method of claim 22 , wherein the concentration of the anti-CD20 antibody in the concentrated anti-CD20 antibody composition is at least 100 mg/ml.

24 . The method of claim 22 , wherein the concentration of the anti-CD20 antibody in the concentrated anti-CD20 antibody composition is about 150 mg/mL to about 250 mg/mL.

25 . The method of claim 22 , wherein the cell culture medium comprises hypotaurine at a concentration from about 2.0 mM to about 50.0 mM.

26 . The method of claim 22 , wherein the cell culture medium comprises:

(i) s-carboxymethylcysteine at a concentration from about 8.0 mM to about 12.0 mM,

(ii) butylated hydroxyanisole at a concentration from about 0.025 mM to about 0.040 mM,

(iii) quercitrin hydrate at a concentration from about 0.010 mM to about 0.020 mM, or

(iv) aminoguanidine at a concentration from about 0.0003 mM to about 10 mM.

27 . The method of claim 22 , wherein the anti-CD20 antibody is an IgG1 monoclonal antibody.

28 . The method of claim 22 , wherein the concentrated anti-CD20 antibody composition is formulated for subcutaneous injection.

Continuity (8)

References Cited (268)