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US 2011/0086774 A1Published application
Protein Detection Via Nanoreporters
AssigneeNanoString Technologies, Inc.
InventorDwayne L. Dunaway
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Abstract
The invention provides methods, compositions, kits and devices for the detection of proteins. In some embodiments, the invention allows for multiplexed protein detection.
Claims57
A method for determining the concentration of at least one protein in a sample comprising the steps:
- (a) providing:
- (i) at least one protein,
- (ii) a first protein probe specific for a first region of said at least one protein, wherein said first protein probe comprises a capture region,
- (iii) a second protein probe specific for a second region of said at least one protein, wherein said second protein probe comprises a nanoreporter comprising a plurality of different detectable labels, and
- (iv) a matrix having attached thereto a moiety which is capable of binding to said capture region in said first protein probe
- (b) forming at least a complex comprising said at least one protein, said first protein probe, said second protein probe and said moiety, wherein said at least one protein is bound to said first and second protein probes, and wherein said moiety is bound to said capture probe in said first protein probe;
- and
- (c) individually detecting said complex or at least part of said complex by a method comprising individually counting the presence of one or more molecules of said nanoreporter wherein the presence of said one or more molecules is indicative of the concentration of said protein in the sample.
5 more elements
A method for determining the concentration of at least one protein in a sample comprising the steps:
- (a) providing:
- (i) at least one protein,
- (ii) a first protein probe specific for a first region of said at least one protein, wherein said first protein probe is attached to a first capture region or a first matrix
- (iii) a second protein probe specific for a second region of said at least one protein, wherein said second protein probe comprises a signal oligo;
- and
- (iv) when said first probe is attached to a first capture region:
- a second matrix having attached thereto a moiety which is capable of binding to said capture region in said first protein probe.
- (b) forming at least a first complex comprising said at least one protein, said first protein probe, and said second protein probe, wherein said at least one protein is bound to said first and second protein probes, and wherein when said first probe is attached to a first capture region said capture probe is bound to said moiety in said second matrix;
- (c) releasing said signal oligo from said first complex;
- (d) forming a second complex comprising (1) at least said signal oligo and (2) at least one oligo probe comprising a signal oligo-specific region and a region comprising a nanoreporter wherein said nanoreporter comprises a plurality of different detectable labels;
- (e) individually detecting said second complex or at least part of said second complex by a method comprising individually counting the presence of one or more molecules of said nanoreporter, wherein the presence of said second one or more molecules is indicative of the concentration of said protein in said sample.
8 more elements
A method for determining the concentration of at least one protein in a sample comprising the steps:
- (a) providing:
- (i) at least one protein,
- (ii) a first protein probe specific for a first region of said at least one protein, wherein said first protein probe is attached to a first oligo
- (iii) a second protein probe specific for a second region of said at least one protein, wherein said second protein probe is attached to a second oligo;
- (b) forming a first complex comprising said at least one protein, said first protein probe and said second protein probe, wherein said at least one protein is bound to said first and second protein probes,
- (c) ligating said first and said second oligo to form a signal oligo
- (d) forming a second complex comprising (1) said first signal oligo and (2) a at least one oligo probe comprising a signal oligo-specific region and a region comprising a nanoreporter wherein said nanoreporter comprises a plurality of different detectable labels
- (e) individually detecting said second complex or at least part of said second complex by a method comprising individually counting the presence of one or more molecules of said nanoreporter, wherein the presence of said one or more molecules is indicative of the concentration of said protein in said sample.
5 more elements
A population of uniquely labeled protein probes, wherein each probe comprises
- i) a target-specific region;
- and
- ii) a region comprising a nanoreporter comprising a plurality of different detectable molecules;
- wherein said nanoreporter in each protein probe has a detectable signal that distinguishes it from other nanoreporters in said population.
1 more element
Dependencies are inferred from the claim text.