Methods and systems for detecting an analyte in a sample

• US 9,523,682 B2
• Filed: 11/16/2012
• Issued: 12/20/2016
• Est. Priority Date: 11/16/2011
• Status: Active Grant

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Huang, Wei, Bornheimer, Scott
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US14/237,858
• Publication Number:
  US 20140170642A1
• Time in Patent Office: 1,495 Days
• US Class Current: 1/1
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2300/0838 Capillaries
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 2021/0346 Capillary cells; Microcells
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 21/05 Flow-through cuvettes G01N21/09 takes precedence; handling fluid samples G01N1/10
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing

Abstract

The present disclosure provides methods for the detection of one or more analytes in a sample. Aspects of the methods include flowing a sample (e.g., a biological sample, such as blood) through a channel comprising an analyte specific capture domain stably associated with a surface thereof, wherein the analyte specific capture domain comprises particles displaying a specific binding member for an analyte; and imaging the analyte specific capture domain to detect whether the analyte is present in the sample. Also provided are systems, devices, and kits that may be used in practicing the subject methods. Methods and compositions as described herein find use in a variety of different applications, including diagnostic applications.

Claims

1. A method of detecting whether an analyte is present in a sample, the method comprising:

(a) contacting a sample with a fluorescent label that specifically binds to the analyte to produce a fluorescently labelled sample;

(b) flowing the fluorescently labelled sample through a capillary channel of a cartridge device comprising a first analyte specific capture domain on an inner surface of the capillary channel at a known location, wherein the first analyte specific capture domain comprises;

(i) particles adhered to the surface of the capillary channel; and

(ii) a binding member that specifically binds to fluorescently labeled analyte of the fluorescently labeled sample, wherein the binding member is contained in and/or on a surface of the particles and wherein the capillary channel has a defined height to particle ratio; and

(c) imaging the first analyte specific capture domain comprising the adhered particles in the capillary channel to detect whether fluorescently labeled analyte is bound by the binding member of the particles and therefore present in the sample, wherein the imaging is performed without washing to remove unbound sample components prior to imaging.

2. The method according to claim 1, wherein the detecting is quantitative.

3. The method according to claim 1, wherein the detecting is qualitative.

4. The method according to claim 1, wherein the capillary channel has a height that is 50 fold greater or less than an average diameter of the particles.

5. The method according to claim 1, wherein the particles are beads.

6. The method according to claim 1, wherein the particles are cells.

7. The method according to claim 1, wherein the sample is a biological sample.

8. The method according to claim 1, wherein the biological sample is from a human.

9. The method according to claim 1, further comprising generating a diagnostic report based on whether the analyte is present in the sample.

10. The method according to claim 1, wherein the particles are associated with an upper surface of the capillary channel and the imaging is conducted from above the capillary channel.

11. The method according to claim 1, wherein the capillary channel comprises a second analyte specific capture domain on an inner surface of the capillary channel at a known location, wherein the second analyte specific capture domain comprises:

(a) particles adhered to the surface of the capillary channel; and

(b) a second binding member that specifically binds to the second analyte that is fluorescently labelled and present in the fluorescently labelled sample, wherein the second binding member is contained in and/or on a surface of the particles; and

wherein the method further comprises imaging the second analyte specific capture domain comprising the adhered particles in the capillary channel to detect whether the second analyte that is fluorescently labelled is bound by the second binding member of the particles and therefore present in the sample.

12. The method according to claim 11, wherein the first and second analyte specific capture domains are located at different positions in the capillary channel.

13. The method according to claim 1, wherein the particles are adhered to the surface of the capillary channel by covalent bonding.

14. The method according to claim 1, wherein the particles are adhered to the surface of the capillary channel by treating the surface of the capillary channel with oxygen plasma to produce a plasma etched surface and depositing the particles on the plasma etched surface in a manner sufficient to stably associate the particles with the inner surface of the capillary channel to produce capture beads.

15. A kit comprising:

a cartridge device having a capillary channel that comprises an analyte specific capture domain on an inner surface of the capillary channel at a known location, wherein the analyte specific capture domain comprises;

(a) particles adhered to the surface of the capillary channel; and

(b) a binding member that specifically binds to the analyte, wherein the binding member is contained in and/or on a surface of the particles and wherein the capillary channel has a defined height to particle ratio; and

a fluorescent label that specifically binds to the analyte when the analyte is bound by the binding member.