End-to-end cell therapy automation

Patent metadata

Field	Value
Publication number	US-11827902-B2
Application number	US-202217751999-A
Country	US
Kind code	B2
Filing date	May 24, 2022
Priority date	Sep 1, 2017
Publication date	Nov 28, 2023
Grant date	Nov 28, 2023

Abstract

The present disclosure provides an automated method of producing genetically modified immune cells, including chimeric antigen receptor T (CAR T) cells, utilizing a fully-enclosed cell engineering system.

First claim

The invention claimed is:

1. A method for automated production of a genetically modified T cell culture, the method comprising:
(a) activating a T cell culture with an activation reagent selected from an antibody and a dendritic cell to produce an activated T cell culture in a first chamber of a fully enclosed cell engineering system;
(b) transducing within the first chamber the activated T cell culture with a viral vector encoding an ectodomain, a transmembrane domain, and an endodomain, to introduce the viral vector into the activated T cell culture and produce a transduced T cell culture;
(c) expanding within the first chamber of the cell engineering system, the transduced T cell culture;
(d) concentrating the expanded T cell culture of (c); and
(e) harvesting the concentrated T cell culture of (d) to produce a genetically modified T cell culture,
wherein (a) through (e) are performed within the fully enclosed cell engineering system,
wherein the fully enclosed cell engineering system is pre-filled with the cell culture, a culture media, the activation reagent, and/or the viral vector, prior to starting the method, and
wherein expansion of the transduced T cell culture in (c) produces at least 20% more genetically modified T cells than expansion utilizing manual cell culture with a flexible, gas permeable bag and transduction efficiency of the method is at least 20% higher than transduction efficiency utilizing manual cell culture with the flexible, gas permeable bag.

2. The method of claim 1 , wherein the cell engineering system is configured to store the culture media, the activation reagent, and/or the viral vector in a low-temperature chamber.

3. The method of claim 2 , wherein the method further includes warming the media before introducing the warmed media to the cell culture.

4. The method of claim 1 , wherein the first chamber is a chamber for growing cells, and wherein the cell engineering system further includes additional chambers for storing the cell culture, the culture media, the activation reagent, and/or the viral vector.

5. The method of claim 1 , wherein the cell engineering system includes multiple reagent bags to enable all reagents required for the process to be pre-loaded and stored.

6. The method of claim 1 , wherein the method process is a self-adjusting process and includes:
(a) monitoring with one or more of a temperature sensor, a pH sensor, a glucose sensor, an oxygen sensor, a carbon dioxide sensor, and an optical density sensor; and
(b) adjusting one or more of a temperature, a pH level, a glucose level, an oxygen level, a carbon dioxide level, and an optical density of the transduced T cell culture, based on the monitoring.

7. The method of claim 1 , wherein the method produces at least about 100 million viable genetically modified T cells.

8. The method of claim 1 , wherein T cell culture is a chimeric antigen receptor T (CAR T) cell culture.

9. The method of claim 1 , wherein the vector encodes a chimeric antigen receptor.

10. The method of claim 1 , wherein the expanding comprises feeding, washing, monitoring, and selecting of the transduced T cell culture.

11. The method of claim 1 , wherein the cell engineering system is capable of performing several rounds of feeding, washing, monitoring, and selecting of the transduced T cell culture.

12. The method of claim 1 , wherein the concentrating comprises centrifugation, supernatant removal following sedimentation, or filtration.

13. The method of claim 1 , further comprising removing the activation reagent from the activated T cell culture after step (a).

14. The method of claim 1 , further comprising removing the viral vector following the transducing in (b).

15. The method of claim 1 , wherein the cell culture comprises at least one accessory cell.

16. The method of claim 15 , wherein the accessory cell is a monocyte, an antigen-presenting cell or a dendritic cell.

17. The method of claim 15 , wherein the accessory cell comprises antigens for a T cell receptor, selected from CD28, CD40, CD2, CD40L, ICOS and combinations thereof.

Assignees

• Lonza Walkersville Inc
• Lonza Cologne Gmbh
• Octane Biotech Inc

Inventors

• Shi Yaling
• Mcafee Erika
• Bandapalle Samatha
• Siehoff Ann
• Gleissner Timo
• O'Connor Joseph
• Abraham Eytan
• Purpura Kelly
• Trainor Nuala
• Smith Timothy

Classifications

• C12M41/44

  of volume or liquid level · CPC title

• C12M41/12

  of temperature (controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00) · CPC title

• A61K40/4205

  Her-2/neu/ErbB2, Her-3/ErbB3 or Her 4/ ErbB4 · CPC title

• A61K40/31

  Chimeric antigen receptors [CAR] · CPC title

• A61K40/11

  T-cells, e.g. tumour infiltrating lymphocytes [TIL] or regulatory T [Treg] cells; Lymphokine-activated killer [LAK] cells · CPC title