Three-dimensional bioreactor for T-cell activation and expansion for immunotherapy

Patent metadata

Field	Value
Publication number	US-11149244-B2
Application number	US-201815945000-A
Country	US
Kind code	B2
Filing date	Apr 4, 2018
Priority date	Apr 4, 2018
Publication date	Oct 19, 2021
Grant date	Oct 19, 2021

Abstract

The present invention relates to the application of a three-dimensional (3D) bioreactor for T-cell expansion for immunotherapy.

First claim

What is claimed is:

1. A method for T-cell expansion comprising:
supplying a 3D bioreactor of biocompatible polymeric material comprising a plurality of voids having a surface area for cell expansion, said plurality of voids having a diameter D, a plurality of pore openings between said voids having a diameter d, such that D>d and wherein: (a) 90% or more of said voids have a selected void volume (V) that does not vary by more than +/−10.0%; and (b) 90% or more of said pore openings between said voids have a value of d that does not vary by more than +/−10.0%;
providing a polydopamine coating on said bioreactor surface area for cell expansion;
providing a tetrameric protein attached to said polydopamine coating;
providing one or more biotinylated antibodies immobilized on said tetrameric protein;
flowing T-cells through said bioreactor having T-cell receptors where said T-cell receptors bind to said one or more biotinylated antibodies and are activated;
exposing the activated T-cells to a perfusion media containing a signaling molecule to promote T-cell expansion.

2. The method of claim 1 wherein said tetrameric protein comprises avidin or streptavidin.

3. The method of claim 1 wherein said biotinylated antibody comprises anti-CD3 antibody.

4. The method of claim 1 wherein said biotinylated antibody comprises anti-CD3 antibody and anti-CD28 antibody.

5. The method of claim 1 wherein said biotinylated antibody comprises anti-CD3 antibody, anti-CD28 antibody, and anti-CD2 antibody.

6. The method of claim 1 wherein said signaling molecule comprises a cytokine signaling molecule.

7. The method of claim 1 wherein said voids have a diameter (D) of 0.4 mm to 100.0 mm.

8. The method of claim 1 wherein said voids have a diameter (D) of 0.4 mm to 25.0 mm.

9. The method of claim 1 wherein said pores have a diameter (d) in the range of 0.2 mm to 10.0 mm.

10. The method of claim 1 wherein 95.0% or more of said voids indicate a void volume (V) that does not vary by more than +/−10.0%.

11. The method of claim 1 wherein 99.0% to 100% of said voids indicate a void volume (V) that does not vary by more than +/−10.0%.

12. The method of claim 1 wherein 95.0% or more of said pore openings between said voids have a value of d that does not vary by more than +/−10.0%.

13. The method of claim 1 wherein 99.0 to 100% or more of said pore openings between said voids have a value of d that does not vary by more than +/−10.0%.

14. The method of claim 1 wherein at least 90.0% of the voids present have 2 pore openings per void.

15. The method of claim 1 wherein at least 90.0% of the voids present have 8 to 12 pore openings per void.

16. The method of claim 1 wherein said voids have an internal concave surface.

17. The method of claim 1 wherein said voids comprise spherical voids.

18. The method of claim 17 wherein said spherical voids have a packing efficiency of greater than 64.0% in a 3D cylindrical space.

19. The method of claim 1 wherein said 3D bioreactor is formed from a material that has a Tensile Modulus of at least 0.01 GPa.

20. The method of claim 1 wherein said 3D bioreactor is formed from a material not susceptible to hydrolysis during cell cultivation such that the amount of hydrolysis does not exceed 5.0% by weight of the material present.

21. The method of claim 1 wherein said bioreactor has a diameter Φ and a height H and the ratio Φ:H is in the range of greater than 1:1 to 100:1.

Assignees

• Southwest Res Inst

Inventors

• Ling Jian

Classifications

• C12N5/0636

  T lymphocytes · CPC title

• C07K2317/75

  Agonist effect on antigen · CPC title

• C12M1/14

  with means providing thin layers or with multi-level trays · CPC title

• C12M25/02

  Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00) · CPC title

• C12M1/005

  Incubators · CPC title