Abvitro LLC

Adrian Wrangham Briggs

Francois Vigneault

Stephen Jacob Goldfless

Brian J Belmont

Sonia Timberlake

Christopher Ryan Clouser

Yue Jiang

Rajagopal Chari

Ronald Hause

Stephen J Goldfless

François Vigneault

Provided herein are methods and composition for immune repertoire sequencing and single cell barcoding. In some aspects, such methods may comprise steps of: (a) forming a plurality of first vessels each comprising: (i) a single cell, and (ii) a single solid support; (b) copying onto the single solid support: (i) a first copy of a first cell polynucleotide from the single cell, and (ii) a second copy of a second cell polynucleotide from the single cell; (c) forming a plurality of second vessels each comprising (i) a single solid support from the plurality of first vessels, and (ii) a barcoded polynucleotide; and (d) amplifying (i) the first copy and the second copy with a first primer set, and (ii) the barcode with a second primer set, wherein a primer of the first primer set is complementary to a primer of the second set; and (e) forming first and second single cell barcoded sequences.

Single cell bar-coding for antibody discovery

Provided herein are methods for target gene sequencing and single cell barcoding in conjunction with analysis of gene expression in single cells. In some embodiments, the target gene is an immune molecule, such as an antibody or TCR. In some embodiments, the methods can be used to carry out transcriptome sequencing, e.g., RNA sequencing, to capture transcriptome of single cells paired with full receptor immune receptor sequences such that information about the immune repertoire and transcriptome of a cell can be determined. Also provided are polynucleotide libraries for use in carrying out transcriptome analysis and immune molecule, e.g., antibody or TCR, sequencing.

High-throughput polynucleotide library sequencing and transcriptome analysis

This invention relates to novel anti-HIV antibodies that can be used in the treatment and detection of human immunodeficiency virus (HIV). These antibodies exhibit a high degree of sensitivity and can provide a broad range of specificity.

Broadly neutralizing anti-HIV-1 antibodies that bind to an N-glycan epitope on the envelope

Provided herein are methods and compositions for single cell characterization using affinity-oligonucleotide conjugates. In some aspects, such methods may comprise attaching a first vessel barcoded polynucleotide to an oligonucleotide portion of an affinity-oligonucleotide conjugate, which binds to a target antigen expressed by a single cell that is isolated in a single vessel. In some aspects, the oligonucleotide portion of the affinity-oligonucleotide conjugate may comprise an antigen identification sequence (AID). In some aspects, the oligonucleotide portion of the affinity-oligonucleotide conjugate may further comprise an affinity molecular barcode (AMB) sequence. In some aspects, such methods may further comprise lysing the single cell and attaching a second vessel barcoded polynucleotide to a cell polynucleotide from the single cell.

Single cell characterization using affinity-oligonucleotide conjugates and vessel barcoded polynucleotides

Provided herein are methods and compositions for single cell characterization using affinity-oligonucleotide conjugates. Provided herein are methods and compositions for single cell charaterization using tetramer-oligonucleotide conjugates.

Affinity-oligonucleotide conjugates and uses thereof

Provided herein are methods and composition for immune repertoire sequencing and single cell barcoding. The methods and compositions can be used to pair any two sequences originating from a single cell, such as heavy and light chain antibody sequences, for antibody discovery, disease and immune diagnostics, and low error sequencing.

Methods of sequencing, determining, pairing, and validating therapeutic agents and disease specific antigens

Provided herein are methods and composition for high-throughput T-cell receptor target identification of natively paired T-cell receptor sequences.

High throughput process for T cell receptor target identification of natively-paired T cell receptor sequences

Provided herein are methods and composition for immune repertoire sequencing and single cell barcoding. The methods and compositions can be used to heavy and light chain antibody sequences originating from a single cell, antibody discovery, disease and immune diagnostics, and low error sequencing.

Provided herein are methods and composition for immune repertoire sequencing and single cell barcoding. The methods and compositions can be used to pair any two sequences originating from a single cell, such as heavy and light chain antibody sequences, alpha and beta chain T-cell receptor sequences, or gamma and delta chain T-cell receptor sequences, for antibody and T-cell receptor discovery, disease and immune diagnostics, and low error sequencing.

High-throughput nucleotide library sequencing

Provided herein are methods and compositions for single cell characterization using affinity-oligonucleotide conjugates, including tetramer-oligonucleotide conjugates (e.g., pMHC-oligonucleotide). In some aspects, such methods may comprise the steps of: forming a vessel comprising an affinity-oligonucleotide conjugate and a T cell; attaching a vessel barcoded polynucleotide or complement thereof to: (a) an oligonucleotide portion of a pMHC-oligonucleotide conjugate, and (b) a T cell receptor (TCR) polynucleotide originating from the T cell; and sequencing the oligonucleotide portion or a complement thereof and the TCR polynucleotide or a complement thereof. Additional methods provided herein include determining the binding affinity of a T cell receptor (TCR) to a pMHC-oligonucleotide. Compositions of the present invention encompass a plurality of vessels, wherein a vessel of the plurality of vessels may comprise a single T cell; and a vessel barcoded polynucleotide, wherein the vessel may further comprise a pMHC-oligonucleotide conjugate.

Methods of selecting T cell receptors using affinity oligonucleotide conjugates

Provided herein are methods for immune repertoire sequencing and single cell barcoding. In some aspects, such methods may comprise producing a plurality of cDNAs from a plurality of polynucleotides, wherein at least one polynucleotide encodes a heavy and/or light chain, from a plurality of immune cells from a biological sample. Producing a plurality of cDNAs may comprise reverse transcription and template switching, such as to form a plurality of uniquely barcoded cDNAs. Methods described herein further comprise amplifying the plurality of uniquely barcoded cDNAs, thereby forming a library, and sequencing one or more of the sequences of the library.

Methods of single-cell barcoding and sequencing

Provided herein are methods and composition for immune repertoire sequencing and single cell barcoding. In some aspects, such methods may comprise steps of: (a) forming a plurality of vessels each comprising a first cell cDNA of a first cell polynucleotide from a single cell and a second cell cDNA of a second cell polynucleotide from the single cell; (b) extending the first cell cDNA that is hybridized to a barcoded polynucleotide comprising a primer binding site sequence and a barcode sequence and a cDNA annealing sequence, or an amplicon thereof; and extending the second cell cDNA that is hybridized to a barcoded polynucleotide comprising a primer binding site sequence and the barcode sequence and a cDNA annealing sequence, or an amplicon thereof; and (c) amplifying the extended first cell cDNA and the extended second cell cDNA with a primer set.

Growing community of inventors

Seattle, WA, United States of America

Adrian Wrangham Briggs