Automated control of cell culture using Raman spectroscopy

Patent metadata

Field	Value
Publication number	US-12352625-B2
Application number	US-202318171438-A
Country	US
Kind code	B2
Filing date	Feb 20, 2023
Priority date	Oct 6, 2017
Publication date	Jul 8, 2025
Grant date	Jul 8, 2025

Abstract

The monitoring and control of bioprocesses is provided. The present disclosure provides the ability to generate generic calibration models, independent of cell line, using inline Raman probes to monitor changes in glucose, lactate, glutamate, ammonium, viable cell concentration (VCC), total cell concentration (TCC) and product concentration. Calibration models were developed from cell culture using two different CHOK1SV GS-KO™ cell lines producing different monoclonal antibodies (mAbs). Developed predictive models, qualified using an independent CHOK1SV GS-KO™ cell line not used in calibration, measured changes in glucose, lactate, ammonium, VCC, and TCC with minor prediction errors over the course of cell culture with minimal cell line dependence. The development of these generic models allows the application of spectroscopic PAT techniques in a clinical manufacturing environment, where processes are typically run once or twice in GMP manufacturing based on a common platform process.

First claim

What is claimed:

1. A method of controlling a bioreactor process, comprising:
supplying a cell culture in a perfusion bioreactor,
wherein conditions in the perfusion bioreactor allow for propagation of the cell culture to produce a product;
measuring one or more parameters within the perfusion bioreactor by a Raman probe to generate Raman spectra,
wherein one or more parameters are selected from the group consisting of nutrient concentration, waste product concentration, glucose concentration, lactate concentration, glutamate concentration, ammonium concentration, viable cell concentration, total cell concentration, product concentration, process conditions, and product characteristics;
measuring a mass of the cell culture within the perfusion bioreactor;
withdrawing one or more of parameter influencing substances from the perfusion bioreactor using one or more ports
wherein the one or more of parameter influencing substances comprises one or more nutrient medias;

adding one or more of parameter influencing substances to the perfusion bioreactor using one or more ports; and
adjusting a flow of one or more of parameter influencing substances to and/or from the perfusion bioreactor in order to maintain one or more parameters within preset limits,
wherein the flow of one or more of parameter influencing substances into and out of the perfusion bioreactor are adjusted based on the Raman spectra and/or the mass of the cell culture of the perfusion bioreactor.

2. A method as defined in claim 1 , wherein the product produced by the cell culture is a protein.

3. A method as defined in claim 1 , wherein one or more parameters are measured periodically or continuously.

4. A method as defined in claim 1 , wherein one or more of parameter influencing substances are withdrawn from the bioreactor continuously or periodically.

5. A method as defined in claim 1 , wherein prior to adjusting the flow of one or more parameter influencing substances, the method comprises:
determining a loop control, based on one or more parameters measured by the Raman probe, that selectively increases or decreases flow of one or more parameter influencing substances to and/or from the bioreactor,
wherein one or more parameters are measured at least every hour.

6. A method as defined in claim 1 , wherein the flow of one or more of parameter influencing substances to and/or from the perfusion bioreactor are adjusted automatically.

7. A method as defined in claim 1 , wherein the method is configured to maintain the cell culture at an average viable cell concentration between about 0.5 million cells per mL to about 35 million cells per mL.

8. A method as defined in claim 1 , wherein the perfusion bioreactor has a volume of about 1 L to about 20 L, and the method is configured to measure the mass of the cell culture within the perfusion bioreactor in association with a load cell.

9. A method as defined in claim 1 , wherein, when one or more parameters deviate from the preset limit, one or more of parameter influencing substances are maintained so as to minimize fluctuations in another one or more of parameter influencing substances.

10. A method as defined in claim 1 , wherein at least one parameter is an amino acid concentration.

11. A method as defined in claim 1 , wherein the method is configured to maintain the glucose concentration between about 1 g/L to about 10 g/L.

12. A method as defined in claim 1 , wherein the method is configured to maintain the lactate concentration less than about 4.5 g/L.

13. A method as defined in claim 1 , wherein the method is configured to maintain the ammonium concentration less than about 0.25 g/L.

14. A method as defined in claim 1 , wherein one or more parameters include the process conditions selected from the group consisting of pH, oxygen tension, dissolved carbon dioxide, temperature, agitation conditions, alkali condition, pressure, and foam levels.

15. A method as defined in claim 1 , wherein one or more parameter influencing substances include a nutrient media comprising a carbohydrate source, an amino acid, a vitamin, a lipid, a protein, a peptide, or mixtures thereof.

16. A method as defined in claim 1 , wherein the product characteristics comprise glycosylation profile and/or charge variant profile.

17. A method as defined in claim 1 , wherein the withdrawing of one or more of parameter influencing substances form the perfusion bioreactor and/or the adding of one or more of parameter influencing substances to the perfusion bioreactor is controlled by a controller that regulates a respective pump.

18. A method as defined in claim 1 , wherein the withdrawing of one or more of parameter influencing substances form the perfusion bioreactor is effectuated using a dip tube.

19. A method as defined in claim 1 , wherein the perfusion bioreactor has a volume of at least 10 L and wherein the glucose concentration is maintained at a desired set point by way of the cell culture fluids being continuously removed and replenished based on measurements by the Raman probe.

20. A method as defined in claim 1 , wherein the method is configured to maintain the cell culture at an average viable cell concentration of at least 30 million cells per mL and wherein one or more parameters deviate from a desired set point within ±2.0 RMSEP.

Assignees

• Lonza Ag

Inventors

• Webster Thaddaeus
• Hadley Brian
• Mason Carrie
• Jaques Colin
• Tummala Seshu
• Rowland-Jones Ruth Christine
• Levinson Yonatan
• Uth Nicholas
• Sinha Pankaj
• Abraham Eytan

Classifications

• G06F16/25

  Integrating or interfacing systems involving database management systems · CPC title

• G01N21/65

  Raman scattering · CPC title

• C12M31/08 Primary

  by conducting or reflecting elements located inside the reactor or in its structure · CPC title

• C12N5/0636

  T lymphocytes · CPC title

• C12N5/0606

  Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696) · CPC title