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Protein expression profiling

  • US 20060166227A1
  • Filed: 07/22/2005
  • Published: 07/27/2006
  • Est. Priority Date: 06/20/2000
  • Status: Abandoned Application
First Claim
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1. A method for detecting one or more analytes, the method comprising (a) bringing into contact one or more analyte samples and one or more arrays, wherein each array comprises a set of analyte capture agents, wherein each analyte capture agent is immobilized on a solid support in a different predefined region of the solid support, wherein each analyte capture agent interacts with an analyte directly or indirectly, (b) prior to, simultaneous with, or following step (a), bringing into contact at least one of the analyte samples and one or more reporter binding primers, wherein each reporter binding primer comprises a specific binding molecule and a rolling circle replication primer, wherein each specific binding molecule interacts with an analyte directly or indirectly, (c) simultaneous with, or following, either or both steps (a) and (b), incubating the analyte samples, the arrays, and the reporter binding primers under conditions that promote interaction of the specific binding molecules, analytes, and analyte capture agents, (d) prior to, simultaneous with, or following step (b), bringing into contact the reporter binding primers, one or more first-stage amplification target circles, one or more second-stage primers, and one or more second-stage amplification target circles, wherein the first-stage amplification target circles each comprise a single-stranded, circular DNA molecule comprising a primer complement portion, wherein the primer complement portion is complementary to at least one of the rolling circle replication primers, wherein the second-stage primers each comprise a first portion and a second portion, wherein the second-stage amplification target circles each comprise a single-stranded, circular DNA molecule comprising a primer complement portion, wherein the primer complement portion is complementary to the second portion of at least one of the second-stage primers, and incubating the reporter binding primers, second-stage primers, and amplification target circles under conditions that promote hybridization between the first-stage amplification target circles and the rolling circle replication primers and between the second-stage primers and the second-stage amplification target circles, (e) following step (d) and prior to, simultaneous with, or following steps (a), (b), or (c), incubating the reporter binding primers, second-stage primers, and amplification target circles under conditions that promote replication of the amplification target circles, wherein replication of the amplification target circles results in the formation of tandem sequence DNA, wherein each second-stage primer can interact with tandem sequence DNA produced from at least one of the first-stage amplification target circles, wherein detection of tandem sequence DNA indicates the presence of the corresponding analytes.

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