Detection of biomolecules in single cells

Abstract

A method for determining the concentration of at least one target analyte in a sample comprising: a. providing at least a binding agent of a first type and a binding agent of a second type, each comprising a label that is unique for the type of binding agent, wherein the binding agent of a first type and the binding agent of a second type specifically bind to a first target analyte, b. determining the binding characteristics of the binding agent of a first type and the binding agent of a second type to the first target analyte, c. providing a sample comprising cells at a known concentration, d. bringing the binding agent of a first type and the binding agent of a second type into contact with the sample, wherein the binding agent of a first type and the binding agent of a second type bind to the first target analyte forming an analyte-binding agent complex (couplexes), e. compartmentalizing single cells of the sample into a plurality of first partitions, f. complementing each first partition comprising a single cell with a lysis buffer, thereby lysing the single cell in each of the first partitions, g. compartmentalizing single analyte-binding agent complexes (couplexes) into a plurality of second partitions, h. performing a bi-component detection method, thereby determining the number and/or absolute concentration of couplexes and binding agents in the second partition, and i. determining the absolute concentration of the first target analyte per cell by taking into account the initial concentration of the cells in the sample in step a., the binding characteristics determined in step b. and the number and/or absolute concentration of couplexes and binding agents determined in step h.

Dependencies are inferred from the claim text.