End-to-end cell therapy automation

Patent metadata

Field	Value
Publication number	US-11371018-B2
Application number	US-201916668745-A
Country	US
Kind code	B2
Filing date	Oct 30, 2019
Priority date	Sep 1, 2017
Publication date	Jun 28, 2022
Grant date	Jun 28, 2022

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 performed by a cell engineering system, comprising:
(a) activating a T cell culture with an antibody to produce an activated T cell culture in a first chamber of the cell engineering system;
(b) transfecting the activated T cell culture, the transfecting comprising:
(i) transferring via a first sterile, closed connection, the activated T cell culture from the first chamber to an electroporation unit;
(ii) electroporating the activated T cell culture with a non-viral vector encoding an ectodomain, a transmembrane domain, and an endodomain, to introduce the vector into the activated T cell culture and produce a transfected T cell culture;
(iii) transferring via a second sterile, closed connection, the transfected T cell culture to the first chamber or to a second chamber of the cell engineering system;

(c) expanding the transfected 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 expansion of the transfected 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.

2. The method of claim 1 , wherein the electroporation unit is located outside of the cell engineering system and connected to the cell engineering system by the first sterile, closed connection and the second sterile, closed connection.

3. The method of claim 1 , wherein the method produces at least about 50 million viable genetically modified T cells.

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

5. The method of claim 4 , wherein the accessory cell comprises a monocyte, an antigen-presenting cell or a dendritic cell.

6. The method of claim 5 , wherein the accessory cell comprises antigens for a T cell receptor, including CD28, CD40, CD40L and/or Inducible T Cell Co-Stimulator (ICOS).

7. The method of claim 1 , wherein the expanding comprises at least one or more of feeding, washing, monitoring, and selecting of the transfected T cell culture.

8. The method of claim 1 , wherein an oxygen level of the transfected T cell culture is optimized for the T cell culture.

9. The method of claim 1 , wherein the cell engineering system recirculates cell culture media through an oxygenation component during one or more of steps (a) to (e).

10. The method of claim 1 , wherein the cell engineering system recirculates nutrients, waste, released cytokines, and/or dissolved gasses during steps (a) to (e).

11. The method of claim 1 , wherein a carbon dioxide level provided by the cell engineering system decreases during step (c).

12. The method of claim 1 , wherein the cell engineering system is configured to perform several rounds of feeding, washing, monitoring, and selecting of the transfected T cell culture.

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

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

15. The method of claim 1 , wherein the cell engineering system contains the cell culture of (a), the antibody, the cell culture medium, and optionally the non-viral vector prior to starting the method.

16. The method of claim 1 , wherein the cell engineering system comprises a plurality of chambers, and wherein each of steps (a) to (e) is performed in a different chamber of the plurality of chambers of the cell engineering system, each of the antibody, the non-viral vector, and a cell culture medium are contained in a different chamber of the plurality of the chambers prior to starting the method, and wherein at least one of the plurality of chambers is maintained at a temperature for growing cells and at least one of the plurality of chambers is maintained at a refrigerated temperature.

17. The method of claim 1 , wherein the genetically modified T cell culture is a chimeric antigen receptor (CAR) T cell culture.

18. The method of claim 1 , wherein the antibody is an anti-CD3 and/or anti-CD28 antibody.

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

• C12N15/85

  for animal cells · CPC title

• C12N2800/107

  for mammalian · CPC title

• C12N2510/00

  Genetically modified cells · CPC title

• C12N2501/515

  CD3, T-cell receptor complex · CPC title

• C12N2501/51

  B7 molecules, e.g. CD80, CD86, CD28 (ligand), CD152 (ligand) · CPC title