Sequential analysis of biological samples

• US 8,822,147 B2
• Filed: 04/27/2012
• Issued: 09/02/2014
• Est. Priority Date: 11/16/2006
• Status: Active Grant

• Current Assignee: Leica Microsystems Gmbh (Danaher Corporation)
• Original Assignee: General Electric Company
• Inventors: Sood, Anup, Montalto, Michael Christopher, Gerdes, Michael, Seel, Maximilian Lewis, Filkins, Robert John
• Primary Examiner(s): Horlick, Kenneth R.
• Assistant Examiner(s): THOMAS, DAVID C
• Application Number:
  US13/457,531
• Publication Number:
  US 20120231967A1
• Time in Patent Office: 858 Days
• Field of Search: None
• US Class Current: 435/6.1
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

Methods for detecting multiple targets in a biological sample are provided. The methods includes contacting the sample with a first probe; physically binding the first probe to a first target; observing a first signal from the first probe; applying a chemical agent to modify the first signal; contacting the sample with a second probe; physically binding the second probe to a second target; and observing a second signal from the second probe. The methods disclosed herein also provide for multiple iterations of binding, observing, signal modification for deriving information about multiple targets in a single sample. An associated kit and device are also provided.

Claims

1. A method of detecting multiple targets in a biological sample, comprising:

(a) contacting the biological sample with a first plurality of probes capable of binding a first group of different targets in the biological sample;

(b) binding the first plurality of probes to their respective targets;

(c) detecting signals from the first plurality of probes;

(d) applying an oxidizing agent after step (c) to modify the signals;

(e) contacting the biological sample after step (d) with a second plurality of probes capable of binding a second group of different targets in the biological sample;

(f) binding the second plurality of probes to their respective targets; and

(g) detecting signals from the second plurality of probes; and

wherein each probe comprises an antibody and a fluorophore.

2. The method of claim 1, wherein the oxidizing agent is hydrogen peroxide.

3. The method of claim 1, wherein the biological sample comprises a tissue section.