Methods for Assessing the Suitability of Transduced T Cells for Administration

• US 20150190428A1
• Filed: 07/12/2013
• Published: 07/09/2015
• Est. Priority Date: 07/13/2012
• Status: Active Grant

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: June, Carl H., Levine, Bruce L., Kalos, Michael D.
• Granted Patent: US 9,572,836 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2239/31 characterized by the route of administration
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464412 CD19 or B4
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 35/02 specific for leukemia
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C12N 5/0636 T lymphocytes
• CPC Class Codes: C12Q 1/04 Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor C12Q1/6897 takes precedence
• CPC Class Codes: C12Q 1/701 Specific hybridization probes
• CPC Class Codes: C12Q 2600/158 Expression markers
• CPC Class Codes: G01N 2333/195 from bacteria
• CPC Class Codes: G01N 2333/37 from fungi
• CPC Class Codes: G01N 2400/50 Lipopolysaccharides; LPS
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses
• CPC Class Codes: G01N 33/56933 Mycoplasma
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743

Abstract

The invention relates to of analyzing vector supernatants useful for transducing T cells destined for administration to a human subject. The invention also related to methods of analyzing transduced T cells destined for administration to a human subject.

Claims

1. A method of analyzing a genetically modified T cell to detect a contaminant, wherein the genetically modified T cell comprises a nucleic acid encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a signaling domain.

2. The method of claim 1, wherein the genetically modified T cell was genetically modified by transduction with a lentiviral vector.

3. The method of claim 1, wherein the contaminant is at least one selected from the group consisting of endotoxin, mycoplasma, replication competent lentivirus (RCL), p24, VSV-G nucleic acid, HIV gag, residual anti-CD3/anti-CD28 coated beads, mouse antibodies, pooled human serum, bovine serum albumin, bovine serum, culture media components, vector packaging cell or plasmid components, a bacterium and a fungus.

4. The method of claim 3, wherein the bacterium is at least one selected from the group consisting of Alcaligenes faecalis, Candida albicans, Escherichia coli, Haemophilus influenza, Neisseria meningitides, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pneumonia, and Streptococcus pyogenes group A.

5. The method of claim 1, wherein the signaling domain is a CD3 zeta signaling domain.

6. The method of claim 1, wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof.

7. The method of claim 6, wherein the antigen-binding fragment is a Fab or a scFv.

8. The method of claim 1, wherein the antigen binding domain binds to a tumor antigen.

9. The method of claim 8, wherein the tumor antigen is associated with a hematologic malignancy.

10. The method of claim 8, wherein the tumor antigen is associated with a solid tumor.

11. The method of claim 8, wherein the tumor antigen is selected from the group consisting of CD19, CD20, CD22, ROR1, mesothelin, CD33/IL3Ra, c-Met, PSMA, Glycolipid F77, EGFRvIII, GD-2, NY-ESO-1 TCR, MAGE A3 TCR, and any combination thereof.

12. The method of claim 1, wherein the costimulatory signaling region comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof.

13. The method of claim 1, wherein the genetically modified T cell is not administered to a human subject because of the identification and quantification of the contaminant.

14. The method of claim 1, wherein the genetically modified T cell is not administered to a human subject because the quantity of the contaminant is excessive as compared with a corresponding control level.