Granted Patent B2

US 12,553,031 · App. 18/668,092 · Granted Feb 17, 2026

Inventors: Michelle R. Simpson-Abelson (Lithia, FL); Matthew J. Frigault (Dorchester, MA); Wayne P. Rothbaum (New York, NY); Michael Weiser (Miami, FL)

Assignee: Iovance Biotherapeutics, Inc.

C12N5/0638A61K39/0011A61K40/11A61K40/42G01N33/5008G01N33/505G01N33/6866A61K2039/5158A61K2039/545A61K2039/55533A61K2239/38A61K2239/48A61K2239/55C12N2501/2302C12N2502/1114C12N2502/30C12N2503/00G01N2333/57G01N2500/10

Abstract

Claims (27)

1 . A method for determining the potency and/or functionality of a tumor infiltrating lymphocyte (TIL) product comprising TIL cells, the method comprising:

(a) measuring interferon gamma (IFN-γ) release from the TIL product by a co-culture assay, wherein the co-culture assay comprises (i) co-culturing TIL cells from the TIL product with U937 target cells, and (ii) detecting IFN-γ released by the TIL cells, wherein the U937 target cells express a major histocompatibility complex (MHC) and the TIL cells express a T cell receptor (TCR), wherein the TIL cells are activated via allogeneic MHC-TCR engagement between the MHC of the U937 target cells and the TCR of the TIL cells during the co-culture assay;

(b) measuring IFN-γ release from the TIL product by a bead-based assay; and

(c) measuring, using a flow cytometry assay, LAG3 + cell content, KLRG1 + cell content, the sum of CD4 + cell content and CD8 + cell content, total viable TCM and TEM cell population, and CD45 + /CD3 + cell content of the TIL product.

2 . The method of claim 1 , wherein the bead-based assay comprises stimulating TIL cells from the TIL product with an anti-CD3 antibody, an anti-CD28 antibody, and/or an anti-CD137 antibody, and detecting IFN-γ released by the TIL cells.

3 . The method of claim 1 , wherein the co-culture assay for IFN-γ release comprises the steps of:

a) co-culturing the U937 target cells with the TIL cells to provide a co-culture;

b) extracting supernatant from the co-culture; and

c) assessing the supernatant for IFN-γ.

4 . The method of claim 3 , wherein the co-culturing is performed for a time period selected from the group consisting of about 6 hours, about 12 hours, about 18 hours, about 24 hours, about 30 hours, about 36 hours, about 42 hours, and about 48 hours.

5 . The method of claim 3 , wherein the ratio of TIL cell:U937 target cell is about 15:4, about 15:2, about 15:1, or about 30:1.

6 . The method of claim 3 , wherein the co-culturing is performed in a cell culture medium comprising IL-2.

7 . The method of claim 6 , wherein the cell culture medium comprises IL-2 at a concentration of about 300 IU/mL.

8 . The method of claim 3 , wherein an HLA blocking antibody is added in the co-culture as a negative control.

9 . The method of claim 8 , wherein the HLA blocking antibody is used at a concentration of about 1-20 μg/mL.

10 . The method of claim 1 , wherein the TIL product is produced by a method comprising:

(i) performing a first expansion by culturing a first population of TILs in a cell culture medium comprising IL-2 to produce a second population of TILs, wherein the first expansion is performed for about 3-14 days to obtain the second population of TILs;

(ii) performing a second expansion by supplementing the cell culture medium of the second population of TILs with additional IL-2, OKT-3, and antigen presenting cells (APCs), to produce a third population of TILs, wherein the second expansion is performed for about 7-14 days to obtain the third population of TILs, wherein the third population of TILs is a therapeutic population of TILs;

(iii) harvesting the therapeutic population of TILs obtained from step (ii);

(iv) transferring the therapeutic population of TILs from step (iii) to an infusion bag; and

(v) cryopreserving the infusion bag.

11 . The method of claim 10 , wherein steps (a)-(c) are performed after cryopreservation.

12 . The method of claim 1 , wherein the TIL product comprises CD8+ TILs and CD4+ TILs.

13 . The method of claim 1 , wherein the co-culture assay for IFN-γ release comprises the steps of:

a) performing at least three different co-cultures of U937 target cells with the TIL cells at different U937 target cell concentrations;

b) extracting supernatants from each of the co-cultures; and

c) assessing the supernatants from each of the co-cultures for IFN-γ secreted from the TIL cells to determine the potency of the TIL product.

Assignments (1)

Continuity (10)

References Cited (400)