Methods for isolating immune binding proteins

Inventors

Mena, Marco Antonio • Emig, Christopher J. • Haliburton, John • Shahi, Payam

Assignees

Augmenta Bioworks Inc

Abstract

Described herein are immune binding proteins and method for obtaining immune binding proteins from genomic or other sources. Also described herein are nucleic acids encoding the immune binding proteins in which the natural multimeric association of chains is maintained in the nucleic acids and the immune binding proteins made therefrom. For example, nucleic acids encoding antibodies that are amplified from a B-cell using the methods described herein maintain the natural pairing of heavy and light chains from the B-cell. This maintenance of pairing (or multimerization) produces libraries and/or repertoires of immune binding proteins that are enriched for useful binding molecules.

Core Innovation

The invention relates to methods of identifying a target cell of interest that comprises an immune binding protein encoded by a plurality of nucleic acids. A bait particle is used that includes an oligonucleotide and an assay probe of interest comprising an antigen that specifically binds to the target cell of interest, and the oligonucleotide comprises a barcode used for identifying the bait particle.

The method identifies the target cell of interest by first isolating a complex formed by the bait particle and the target cell of interest. The bait particle is identified by sequencing the barcode from the complex, the target cell of interest is then lysed from the complex, and after lysing the plurality of nucleic acids encoding the immune binding protein released from the target cell of interest are sequenced to identify the target cell of interest.

The document further discloses methods and nucleic acids for generating immune binding protein libraries while preserving native in vivo multimeric associations, particularly heavy/light chain pairing from B cells and plasma cells. Library generation and amplification are supported by spatial confinement and sequencing-based identification, including the use of barcoding and identification workflows linked to immune binding protein expression.

Claims Coverage

Independent claim clm-00001 covers a cell-identification workflow with an antigen-bearing bait particle that contains a barcode, followed by sequencing of the barcode to identify the bait and sequencing of immune-binding-protein-encoding nucleic acids after lysis. The inventive features center on antigen-specific bait capture, bait identification by barcode sequencing, and post-lysis sequencing of immune binding protein-encoding nucleic acids to identify the target cell.

Across the independent claim, identification of a target cell of interest is achieved by antigen-specific binding of a barcode-bearing bait particle, barcode sequencing from the bait–target complex, and subsequent sequencing of immune binding protein-encoding nucleic acids released after lysis.

Stated Advantages

Documented Applications

No documented applications found