Granted Patent B2

US 12,584,113 · App. 16/722,541 · Granted Mar 24, 2026

Inventors: Gregory Walter Hiller (Wakefield, MA); Bhanu Chandra Mulukutla (Lawrence, MA)

Assignee: Pfizer Inc.

C12N5/0682C12N5/0018C12N2500/32C12N2500/33C12N2500/34C12N2501/80C12N2501/999

Abstract

Claims (15)

1 . A method of cell culture comprising

(i) providing Chinese hamster ovary (CHO) cells in a cell culture medium and starting a fed-batch cell culture process with the cells in the cell culture medium,

wherein the cell culture medium comprises histidine at a concentration above 2 mM,

wherein the concentration of serine is at a concentration below 1 mM at the start of the culture process, and

wherein the fed-batch cell culture process comprises adding more serine to the cell culture medium at one or more times after starting the cell culture process and,

(ii) maintaining serine below a concentration of 1 mM in the cell culture medium and

wherein step (ii) comprises a step of measuring the concentration of serine in the cell culture medium prior to adding more serine to the cell culture medium and based on the measured concentration of serine in the cell culture medium, adding an amount of serine to the cell culture medium such that after adding the amount of serine, the concentration of serine is below 1 mM in the cell culture medium.

2 . The method of claim 1 , wherein the concentration of phenylalanine, tyrosine, tryptophan, methionine, and glycine is maintained below 1 mM in the cell culture medium.

3 . The method of claim 1 , wherein the cell culture medium comprises one or more of valine, isoleucine, proline, lysine, arginine, aspartate, glutamate and asparagine at a concentration above 1 mM, 1.5 mM, 2 mM, 3 mM or 5 mM.

4 . The method of claim 1 , wherein the cells express a recombinant protein.

5 . The method of claim 1 , wherein the cell culture method comprises a growth phase and a production phase and step (ii) is applied during the growth phase.

6 . The method of claim 1 , wherein the step of measuring the concentration of serine is performed every 30 minutes, every hour, or every two hours.

7 . The method of claim 1 , wherein measuring the concentration of serine is performed using a Raman spectroscopy method.

8 . The method of claim 7 , wherein the Raman spectroscopy method uses an empirical model built by employing a training set of spectral data generated using known concentrations of serine in a mix of spent media samples taken from different time points of different cell culture processes.

9 . The method of claim 1 , wherein the cell culture medium comprises histidine at a concentration above 3 mM or 5 mM.

Assignments (1)

Continuity (3)

References Cited (41)