Filtration-based microarray chip

• US 20030108949A1
• Filed: 07/03/2002
• Published: 06/12/2003
• Est. Priority Date: 07/03/2001
• Status: Abandoned Application

• Current Assignee: Georgia Tech Research Corporation (University System of Georgia)
• Original Assignee: Georgia Tech Research Corporation (University System of Georgia)
• Inventors: Bao, Gang, Xu, Yangqing
• Application Number:
  US10/189,923
• Publication Number:
  US 20030108949A1
• Time in Patent Office: Days
• US Class Current: 435/7.1
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00668 Two-dimensional arrays within three-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00731 Saccharides
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 3/50255 Multi-well filtration
• CPC Class Codes: B01L 9/523 for multisample carriers, e.g. used for microtitration plates
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 40/12 : Libraries containing saccharides or polysaccharides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G01N 1/40 : Concentrating samples
• CPC Class Codes: G01N 1/4077 : by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54386 : Analytical elements
• CPC Class Codes: G01N 33/6803 : General methods of protein analysis not limited to specific proteins or families of proteins

Abstract

Filtration-based microarray chips are provided. The microarray chips comprise a random oriented microporous filtration substrate of charged cellulose esters, such as nitrocellulose, and a plurality of different analyte-specific capture molecules attached to the substrate in a microarray. The capture molecules can be proteins, antigens, monoclonal or polyclonal antibodies, or apatomers. In particular, apparati and methods for the detection of a subject analyte comprising multiple stacked filtration-based microarray chips are also provided. The filtration-based microarray chips permits a sensitive, specific, fast and high throughput assay that can be used to detect analytes associated with a disease.

Claims

1. A filtration-based microarray chip comprising,

a. a random oriented microporous filtration substrate having a planar aspect and comprising charged cellulose esters, and

b. a plurality of different analyte-specific capture molecules attached to the substrate in a microarray, wherein the filtration-based microarray chip permits a fluid solution to flow therethrough and at least a portion of analytes to be captured thereto.

2. The microarray chip of claim 1, wherein the charged cellulose esters are cellulose nitrate, cellulose actetate or mixtures thereof.

3. The microarray chip of claim 1, wherein the charged cellulose esters are nitrocellulose.

4. The microarray chip of claim 1, wherein the filtration substrate micropores are between about 0.05 and 10 microns in diameter.

5. The microarray chip of claim 1, wherein the analyte is a protein.

6. The microarray chip of claim 1, wherein the analyte is an antibody.

7. The microarray chip of claim 1, wherein the analyte is obtained from a sample of bodily fluid.

8. The microarray chip of claim 1, wherein the analyte is obtained from a sample of cell lysate.

9. The microarray chip of claim 1, comprising more than ten different analyte-specific capture molecules attached to the substrate in a microarray.

10. The microarray chip of claim 1, wherein the analyte-specific capture molecules are proteins.

11. The microarray chip of claim 1, wherein the analyte-specific capture molecules are antibodies.

12. The microarray chip of claim 1, wherein the analyte-specific capture molecules are apatomers.

13. An apparatus for analyte detection comprising a plurality of filtration-based microarray chips of any of claims 1-12 aligned with planar aspects in parallel such that a solution flows through the plurality of chips.

14. The apparatus of claim 13, further comprising a holder for the chips and a means for washing a solution of analytes repeatedly through the plurality of chips.

15. The apparatus of claim 14, wherein the means for washing a solution of analytes repeatedly through the plurality of microarray chips is a syringe in fluid communication with a chamber housing the chip holder and the aligned chips.

16. A method of detecting a subject analyte, comprising combining the micrarray chip of claim 1 with a sample suspected of containing the subject analyte, and detecting capture of the analyte on the substrate to determine the presence of the subject protein in the sample.

17. The method of claim 16, wherein the detection of the subject analyte indicates the presence of a disease marker in the sample.

18. The method of claim 16, wherein the analyte is obtained from a sample of bodily fluid.

19. The method of claim 16, wherein the analyte is obtained from a sample of cell lysate.

20. The method of claim 16, wherein detection of the analyte is achieved by labeling the analyte before capture.

21. The method of claim 16, wherein the analyte is labeled with a fluorescent dye.

22. The method of claim 21, wherein detection of the analyte is achieved by labeling the analyte after capture with a labeled antibody.

23. The method of claim 21, wherein the antibody is labeled with a fluorescent dye.

24. The method of claim 16, wherein the detection is performed with single- or multiple-photon microscopy, or time-resolved fluorescence microscopy.