US 2025/0154494 A1 · Microfluidic Co-Encapsulation Device and System and Methods for Identifying T-Cell Receptor Ligands

Abstract

The present invention provides a platform for co-culturing T cells with APC cells expressing a library of antigenic sequences is disclosed, as well as compositions for use in the system, a co-culturing device and methods of use of the system for identifying novel T cell receptor:antigen interactions. The present invention also provides microfluidic co-encapsulation devices configured to generate longitudinal flows of individual particles (such as individual cells) from fluid particle suspensions, combining two or more individual particle flows into a single stream of individual particles, and segmenting the single stream using an isolation fluid, resulting in a suspension of co-encapsulated individual particles from each fluid particle suspension. The present invention also provides methods of using the microfluidic co-encapsulation devices and methods of modifying fluid particle suspensions to promote individual particle separation.

Claims31

Dependent claims

A method of identifying an antigenic polypeptide as a ligand for binding by a T cell receptor (TCR), the method comprising:

a) co-culturing at least one cell comprising:
i) at least one TCR and
ii) a nucleic acid molecule comprising a TCR responsive promoter operably linked to an expression cassette comprising a nucleotide sequence encoding a marker,
with one or more antigen presenting cell (APC) of an APC library comprising a plurality of APCs, wherein each APC comprises a nucleic acid molecule of a minigene library comprising a plurality of nucleic acid molecules wherein each nucleic acid molecule comprises a nucleotide sequence encoding at least one antigenic polypeptide for presentation as an antigenic polypeptide-human leukocyte antigen (HLA) complex, wherein the sequence encoding the at least one antigenic polypeptide is of a predetermined length;
wherein binding of the TCR to at least one antigenic polypeptide-HLA complex of the APC induces expression of the nucleic acid molecule encoding the marker;
b) isolating the APC cell bound to the cell comprising the TCR;
and
c) sequencing the nucleic acid molecule of the minigene library of the APC cell to identify the antigenic polypeptide as a ligand for binding by the TCR.

5 more elements

A nucleic acid molecule encoding an antibody or fragment thereof specific for binding to an antigen-presenting cell (APC) marker selected from the group consisting of CD19 and CD20, wherein the nucleic acid molecule comprises at least one selected from the group consisting of:

a) a nucleotide sequence encoding a heavy chain variable region of an anti-CD19 synthetic antibody selected from the group consisting of SEQ ID NO:4 and SEQ ID NO:12;
b) a nucleotide sequence encoding a light chain variable region of an anti-CD19 synthetic antibody selected from the group consisting of SEQ ID NO:8 and SEQ ID NO:16;
c) a nucleotide sequence encoding a heavy chain variable region of an anti-CD20 synthetic antibody selected from the group consisting of SEQ ID NO:20 and SEQ ID NO:28;
and
d) a nucleotide sequence encoding a light chain variable region of an anti-CD20 synthetic antibody selected from the group consisting of SEQ ID NO:24 and SEQ ID NO:32.

2 more elements

Dependencies are inferred from the claim text.

Microfluidic Co-Encapsulation Device and System and Methods for Identifying T-Cell Receptor Ligands

Life of this patent

Microfluidic Co-Encapsulation Device and System and Methods for Identifying T-Cell Receptor Ligands

Life of this patent

Abstract

Claims31