Automated instrumentation for production of T-cell receptor peptide libraries

Patent metadata

Field	Value
Publication number	US-11293117-B2
Application number	US-202017016454-A
Country	US
Kind code	B2
Filing date	Sep 10, 2020
Priority date	Apr 24, 2018
Publication date	Apr 5, 2022
Grant date	Apr 5, 2022

Abstract

The present disclosure provides instrumentation and automated methods for creating cell surface display libraries, where the cells of the library display engineered peptides on their cell surfaces for identification of antigens that bind to T-cell receptors. The engineered peptides may be putative antigens or binding regions of the T-cell receptors.

First claim

We claim:

1. A method for identifying cells expressing T-cell receptors (TCR's) on their surface comprising:
providing a population of cells;
processing the population of cells using an instrument for multiplexed nuclease-directed genome editing using introduced nucleic acids and a nucleic acid-directed nuclease to create cells, wherein the introduced nucleic acids comprise nucleic acids derived from a T-cell receptor loci;
incubating the processed cells to facilitate nucleic acid editing in the cell; and
allowing the cells to express and display the engineered T-cell receptor on the surface of the cells;
screening the edited cells displaying engineered peptide antigens against a library of antigens.
identifying the edited cells expressing the engineered T-cell receptor that selectively bind to one or more antigens in the library of antigens.

2. The method of claim 1 , wherein the T-cell receptor loci is a TCRα loci.

3. The method of claim 1 , wherein the T-cell receptor loci is a TCRβ loci.

4. The method of claim 1 , wherein the T-cell receptor loci is a TCRδ loci.

5. The method of claim 1 , wherein the T-cell receptor loci is a TCRγ loci.

6. The method of claim 1 , wherein the library of antigens comprises binding regions of a target of interest.

7. The method of claim 6 , wherein computational method are used to pre-select a list of binding regions of the target of interest.

8. The method of claim 7 , wherein the target of interest is a disease related target.

9. The method of claim 1 , wherein the nuclease is an RNA-directed nuclease.

10. The method of claim 9 , wherein the RNA-directed nuclease is selected from MAD7, Cas 9, Cas12/Cpf1.

11. The method of claim 1 , wherein the cells are mammalian cells.

12. A cell library produced using the method of claim 1 .

13. The method of claim 10 , wherein the RNA-directed nuclease is MAD7.

14. The method of claim 10 , wherein the RNA-directed nuclease is Cas9.

15. The method of claim 10 , wherein the RNA-directed nuclease is Cas12/Cpf1.

Assignees

• Inscripta Inc

Inventors

• Federowicz Stephen
• Church Deanna
• Graige Michael

Classifications

• C07K14/70539 Primary

  MHC-molecules, e.g. HLA-molecules · CPC title

• A61K39/00

  Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53) · CPC title

• C40B70/00

  Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes · CPC title

• C40B50/06 Primary

  Biochemical methods, e.g. using enzymes or whole viable microorganisms · CPC title

• C40B40/10

  Libraries containing peptides or polypeptides, or derivatives thereof · CPC title