Method and apparatus for high density format screening for bioactive molecules

• US 6,790,652 B1
• Filed: 01/08/1999
• Issued: 09/14/2004
• Est. Priority Date: 01/08/1998
• Status: Expired due to Term

• Current Assignee: Fisher Bioimage APS
• Original Assignee: BioImage A/S (Fisher Bioimage APS)
• Inventors: Thastrup, Ole, Terry, Bernard Robert, Scudder, Kurt Marshall, Arkhammer, Per Olaf Gunnar
• Primary Examiner(s): Chin, Christopher
• Assistant Examiner(s): GABEL, GAILENE
• Application Number:
  US09/227,518
• Time in Patent Office: 2,076 Days
• Field of Search: 435/4, 435/6, 435/7.1, 435/7.21, 435/7.72, 435/7.92, 435/7.8, 435/177-180, 435/70.1, 435/182, 435/242, 435/325, 435/395, 435/401, 435/286.1, 435/287.1, 435/287.7, 435/7.2, 436/501, 436/528, 436/529-531, 436/546, 436/164, 436/169, 436/170, 436/174, 436/177, 436/800, 436/809, 436/815, 436/34, 436/55, 436/147, 436/162, 436/165, 436/172, 422/51, 422/52, 422/50, 422/55, 422/56, 422/58, 422/60, 422/69, 422/81, 422/82.05, 422/82.07, 422/68.1, 422/101
• US Class Current: 435/287.7
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5079 Mitochondria
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/815 Test for named compound or class of compounds
• CPC Class Codes: Y10T 436/12 Condition responsive control
• CPC Class Codes: Y10T 436/25 including sample preparation

Abstract

A method and apparatus for screening an array of test compounds for bioactivity by contacting an array of test compounds with a detector layer capable of detecting bioactivity, and detecting a detector layer response. The detector layer is comprised of physiologically viable cells. The method and apparatus allow a large number of test compounds to be simultaneously assayed in parallel without the need for complex fluidic devices.

Claims

1. A method for simultaneously exposing an array of test compounds to a detector layer of physiologically viable cells, comprising:

(a) providing an array of test compounds, wherein the test compounds are disposed on a support;

(b) providing a porous membrane, wherein the porous membrane is constructed of a non-absorbent material with pores of regular and defined diameter which traverse the membrane directly from the upper to the lower side;

(c) bringing the array of test compounds in close apposition with the detector layer so that the porous membrane is in contact with a liquid layer surrounding the detector layer and the porous membrane is in contact with the array of test compounds thereby allowing diffusion of the test compounds through the porous membrane to the detector layer.

2. The method according to claim 1, wherein the support is a non-porous substrate.

3. The method according to claim 1, wherein the physiologically viable cells form a monolayer.

4. The method according to claim 1, wherein the physiologically viable cells are supported by an optically clear substrate.

5. The method according to claim 1, wherein the detector layer is held stationary in the field of view of an optical detector and the array of test compounds is moved into contact with said detector layer during the course of measurement.

6. The method according to claim 1, wherein the array of test compounds is held stationary in the field of view of an optical detector and the detector layer is moved into contact with said array of test compounds during the course of measurement.

7. The method according to claim 1, wherein the array of test compounds is generated on the support by combinatorial chemistry.

8. A method for screening test compounds for bioactivity by simultaneously exposing an array of test compounds to a detector layer of physiologically viable cells, comprising:

(a) providing an array of test compounds, wherein each compound is disposed on a support;

(b) providing a porous membrane, wherein the porous membrane is constructed of a non-absorbent material with pores of regular and defined diameter which traverse the membrane directly from the upper to the lower side;

(c) bringing the array of test compounds in close apposition with the detector layer so that the porous membrane is in contact with a liquid layer surrounding the detector layer and the porous membrane is in contact with the array of test compounds thereby allowing diffusion of the test compounds through the porous membrane to the detector layer; and

(d) detecting a response of the detector layer to the test compound.

9. The method according to claim 8, wherein the response is recorded by a sequence of images.

10. The method according to claim 8, wherein the detected response is a change in a luminescence property of the physiologically viable cells in the detector layer.

11. The method according to claim 8, wherein the detected response is a change in a fluorescence property of the physiologically viable cells in the detector layer.

12. The method according to claim 8, wherein the support is a non-porous substrate.

13. The method according to claim 8, wherein the physiologically viable cells form a monolayer.

14. The method according to claim 8, wherein the physiologically viable cells are supported by an optically clear substrate.

15. The method according to claim 8, wherein the detector layer is held stationary in the field of view of an optical detector and the array of test compounds is moved into contact with said detector layer during the course of measurement.

16. The method according to claim 8, wherein the array of test compounds is held stationary in the field of view of an optical detector and the detector layer is moved into contact with said array of test compounds during the course of measurement.

17. The method according to claim 8, wherein the array of test compounds is generated on the support by combinatorial chemistry.