Quantitative massively parallel proteomics
AssigneeVerily Life Sciences LLC
InventorsIan Peikon, Gary Tong, Daniel Liu, Ci Chu
Life of this patent
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Abstract
Provided herein are compositions and methods for the identification of an expression profile in a single cell or population of cells. Kits for use with the disclosed methods are also provided, including antibodies, with a unique molecular identifier and antibody identifier, and primers for amplification of the antibody identifier sequence.
Claims20
A method of identifying an expression profile in a single cell of a population of cells, comprising:
- a) contacting a population of cells with a plurality of antibodies under conditions that promote specific binding of the antibodies to target antigens of the cells, wherein each antibody in the plurality is capable of binding to a unique target antigen in one or more cells, wherein each antibody is conjugated to an oligonucleotide, and wherein the oligonucleotide comprises a unique molecular identifier sequence and an antibody identifier sequence corresponding to the unique target antigen;
- b) separating single cells of the population into individual compartments;
- c) amplifying the unique molecular identifier sequences and antibody identifier sequences in each individual compartment;
- and
- d) analyzing the amplified unique molecular identifier sequences and antibody identifier sequences to identify the expression profile of each single cell.
A method of identifying an expression profile of a population of cells, comprising:
- a) contacting a population of cells with a plurality of antibodies under conditions that promote specific binding of the antibodies to the cells;
- wherein each antibody in the plurality is capable of binding to a unique target antigen in one or more cells, wherein each antibody is conjugated to an oligonucleotide, and wherein the oligonucleotide comprises a unique molecular identifier sequence and an antibody identifier sequence corresponding to the unique target antigen;
- b) amplifying the unique molecular identifier sequences and antibody identifier sequences;
- and
- c) analyzing the amplified unique molecular identifier sequences and antibody identifier sequences to identify the expression profile of the population of cells.
A method of identifying an expression profile of a population of cells, comprising:
- a) lysing a population of cells;
- b) labeling the cellular proteins in the cell lysate of step a) with an affinity label;
- c) contacting the affinity labeled proteins with beads comprising an agent that binds the affinity label to generate protein-coated beads;
- d) contacting the protein-coated beads with a plurality of antibodies under conditions that promote specific binding of the antibodies to the protein-coated beads;
- wherein each antibody in the plurality binds to a unique target protein, wherein each antibody is conjugated to an oligonucleotide, and wherein the oligonucleotide comprises a unique molecular identifier sequence and an antibody identifier sequence corresponding to the unique target protein;
- e) amplifying the unique molecular identifier sequences and antibody identifier sequences;
- and
- f) analyzing the amplified unique molecular identifier sequences and antibody identifier sequences to identify the expression profile of the population of cells.
Dependencies are inferred from the claim text.