Microfabricated, flowthrough porous apparatus for discrete detection of binding reactions

• US 6,893,816 B1
• Filed: 04/21/1998
• Issued: 05/17/2005
• Est. Priority Date: 10/28/1993
• Status: Expired due to Fees

• Current Assignee: UT-Battelle LLC
• Original Assignee: Houston Advanced Research Center
• Inventors: Beattie, Kenneth L.
• Primary Examiner(s): SAUNDERS, DAVID A
• Application Number:
  US09/063,356
• Time in Patent Office: 2,583 Days
• Field of Search: 435/6, 435/7.1, 435/7.2, 435/7.8, 435/970, 435/973, 435/287.1, 435/287.2, 435/288.4, 436/518, 436/527, 436/535, 436/805, 436/807, 436/808, 436/809, 436/501, 422/68.1, 422/69
• US Class Current: 435/6.12
• 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 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00414 using suction
• CPC Class Codes: B01J 2219/00418 using pressure
• CPC Class Codes: B01J 2219/00423 using filtration, e.g. through porous frits
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00574 radioactive
• CPC Class Codes: B01J 2219/00576 fluorophore
• 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 2219/00691 using robots
• CPC Class Codes: B01J 2219/00704 integrated with the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2300/0819 Microarrays; Biochips
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/50255 : Multi-well filtration
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6874 : involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: C12Q 2565/631 : being a biochannel or pore
• CPC Class Codes: C40B 60/08 : Integrated apparatus specially adapted for both creating and screening libraries
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: Y10S 435/97 : Test strip or test slide
• CPC Class Codes: Y10S 435/973 : Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10S 436/807 : Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/808 : Automated or kit
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays

Abstract

An improved microfabricated apparatus for conducting a multiplicity of individual and simultaneous binding reactions is described. The apparatus comprises a substrate on which are located discrete and isolated sites for binding reactions. The apparatus is characterized by discrete and isolated regions that extend through said substrate and terminate on a second surface thereof such that when a test sample is applied to the substrate, it is capable of penetrating through each such region during the course of said binding reaction. The apparatus is especially useful for sequencing by hybridization of DNA molecules.

Claims

1. A method for detecting a binding reaction between a binding reagent and a binding target, comprising:

(A) contacting a sample suspected of containing a binding target with a substrate comprised of;

(i) oppositely facing first and second major surfaces, (ii) a multiplicity of discrete channels extending through said substrate from said first major surface to said second major surface, (iii) at least a first binding reagent immobilized on the walls of at least a first group of said channels and (iv) at least a second binding reagent immobilized on the walls of at least a second group of said channels; and

(B) detecting binding between a binding target in the sample and at least one binding reagent on the walls of at least one group of discrete channels in the substrate, thereby detecting said binding reaction.

2. A method according to claim 1, wherein the first and second binding reagents differ from one another.

3. A method according to claim 1, wherein the first and second binding reagents bind different binding targets.

4. A method according to claim 1, wherein said substrate comprises groups of channels having areas of between about 20 μ

m² to about 3×

10⁶ μ

m².

5. A method according to claim 1, wherein there are between 400 and 4400 of said groups of discrete channels per cm² of cross-sectional area of the substrate.

6. A method according to claim 1 wherein a detectable label is used to detect the reaction.

7. A method according to claim 6, wherein said detectable label is selected from the group consisting of fluorescent, chemiluminescent and radioactive labels.

8. A method according to claim 7, wherein said detectable label is attached to said binding target.