Systems and methods for the detection of biomarkers

• US 8,318,641 B2
• Filed: 10/18/2010
• Issued: 11/27/2012
• Est. Priority Date: 02/10/2010
• Status: Expired due to Fees

• Current Assignee: Companion Diagnostics, Inc.
• Original Assignee: Companion Diagnostics, Inc.
• Inventors: Selinfreund, Richard H., Vig, Rakesh, Gill, Richard P.
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): COLE, MONIQUE T
• Application Number:
  US12/906,235
• Publication Number:
  US 20110195873A1
• Time in Patent Office: 771 Days
• Field of Search: 422/400, 422/69
• US Class Current: 506/39
• CPC Class Codes: A61B 10/0045 Devices for taking samples of body liquids devices for taking blood samples A61B5/15
• CPC Class Codes: C12Q 1/40 involving amylase
• CPC Class Codes: G01N 33/50 Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/105831 Protein or peptide standard or control [e.g., hemoglobin, etc.]
• CPC Class Codes: Y10T 436/2525 Stabilizing or preserving

Abstract

Systems and methods for the detection of biomarkers. In at least one embodiment of a microarray system of the present disclosure, the microarray system comprises a microarray product comprising at least 100 diagnostic markers/cm², a microarray identifier, and a stabilizing agent, a control microarray product comprising a first specific binding pair member that binds to a first detectable label, and a processor for providing information regarding the identification and concentration of markers on the microarray based on the identity of the array provided by the microarray identifier.

Claims

1. A microarray system for diagnosing a disease in a bodily fluid, comprising:

a microarray product comprising at least 100 diagnostic protein markers/cm²,

a labeled microarray identifier, and

a plurality of a stabilizing agent present in an amount or concentration sufficient for stabilizing said at least 100 diagnostic protein markers/cm²;

a control microarray product, located on at least one microarray site bound to said microarray product, and comprising a first specific binding pair member that binds to a first detectable label; and

operably connected to a computer processor;

wherein said processor is programmed for providing information to a computer data base recording the identification and concentration of protein markers on the microarray based on the identity of the array provided by the labeled microarray identifier.

2. The microarray system of claim 1, wherein the microarray product is comprised of a compound selected from the group consisting of gel, nitrocellulose, nylon, quartz, glass, metal, silica based materials, silica, resins, polymers, or combinations thereof.

3. The microarray system of claim 1, wherein the diagnostic protein marker comprises a peptide fragment having a length of at least about 50 amino acids.

4. The microarray system of claim 1, wherein the stabilizing agent is a protease inhibitor.

5. The microarray system of claim 1, wherein the diagnostic marker is selected from the group consisting of a protein, a glycoprotein, an enzyme, an enzyme inhibitor, and a metabolite.

6. The microarray system of claim 1, wherein said plurality of the stabilizing agent is present in an amount or concentration sufficient to completely or substantially inactivate or reduce the activity of an enzyme selected from the group consisting of an amylase, a lysozyme, a peroxidase, a glycosidase, an esterase, a protease, and a peptidase.

7. The microarray system of claim 1, wherein the microarray system is capable of analyzing a body fluid is selected from the group consisting of saliva, a mucous secretion, tears, sweat, semen, urine, a vaginal secretion, exhalate, blood, serum, and an anal secretion.

8. The microarray system of claim 1, wherein the stabilizing agent is selected from the group consisting of citric acid sodium hydroxide buffer pH 6 (Fixanal®

Buffer 6.0 (Sigma-Aldrich Co.)), acetic acid, aluminum hydroxide bentonite, aluminum sulfate hydrate, aluminum potassium sulfate dodecahydrate, benzoic acid, caffeine, and 3-tert-butyl-hydroxyanisole, or a combination thereof.

9. The microarray system of claim 1, wherein the plurality of stabilizing agents each present in approximately the same concentration.

10. The microarray system of claim 1, wherein said plurality of the stabilizing agent is capable of inhibiting degradation of the diagnostic protein marker to an inhibitory degree, wherein the inhibitory degree is selected from the group consisting of at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, and at least about 99%.

11. The microarray system of claim 1, wherein said plurality of the stabilizing agent has a concentration selected from the group consisting of about 200 parts per million (ppm) to about 2000 ppm, about 400 ppm to about 1600 ppm, about 600 ppm to about 1400 ppm, about 800 ppm to about 1200 ppm, and about 400 ppm to about 600 ppm.

12. The microarray system of claim 1, wherein said plurality of the stabilizing agent is able to inhibit the degradation or inactivation of the diagnostic marker for an inhibitory period selected from the group consisting of at least one minute, at least about five minutes, at least about ten minutes, at least about fifteen minutes, at least about thirty minutes, at least about one hour, at least about two hours, at least about four hours, and at least about eight hours.

14. The microarray system of claim 1 for diagnosing a disease in a bodily fluid, comprising:

a filter membrane cover to prevent body fluid particulates from contaminating said microarray product.

13. A microarray system for diagnosing a disease in a bodily fluid, comprising:

a microarray product comprising at least 100 diagnostic markers/cm²,

a filter membrane cover effectively preventing body fluid particulates from contaminating said microarray product,

a labeled microarray identifier, and

a plurality of a stabilizing agent present in an amount or concentration sufficient for stabilizing said at least 100 diagnostic protein markers/cm², selected from the group consisting of citric acid sodium hydroxide buffer pH 6 (Fixanal®

Buffer 6.0 (Sigma-Aldrich Co.)), acetic acid, aluminum hydroxide bentonite, aluminum sulfate hydrate, aluminum potassium sulfate dodecahydrate, benzoic acid, caffeine, and 3-tert-butyl-hydroxyanisole, or a combination thereof;

a control microarray product, located on at least one microarray site bound to said microarray product, and comprising a first specific binding pair member that binds to a first detectable label; and

operably connected to a computer processor;

wherein said

processor is programmed for providing information to a computer data base recording an identification and concentration of protein markers on the microarray based on the identity of the array provided by the labeled microarray identifier; and

the database in communication with the processor containing at least one correlation between the diagnostic protein marker and a disease state.

15. The microarray system of claim 13 for diagnosing a disease in a bodily fluid, wherein the labeled microarray identifier comprises a specific protein marker specific probe selected from a high affinity antibody, an antibody fragment or a mixture thereof.