Biosensor arrays and methods

• US 6,699,719 B2
• Filed: 07/22/2002
• Issued: 03/02/2004
• Est. Priority Date: 11/29/1996
• Status: Expired due to Term

• Current Assignee: Proteomic Systems Incorporated
• Original Assignee: Proteomic Systems Incorporated
• Inventors: Groves, John T., Ulman, Morrison, Boxer, Steven G., Yamazaki, Miki (Vikky), Nguyen, Luat T.
• Primary Examiner(s): SNAY, JEFFREY R
• Application Number:
  US10/200,682
• Publication Number:
  US 20030096418A1
• Time in Patent Office: 589 Days
• Field of Search: 422/68.1, 422/82.02, 422/82.06, 436/71, 436/525, 436/527, 436/528, 436/164, 436/172, 435/287.2, 435/287.1, 427/2.11, 427/2.13
• US Class Current: 506/30
• 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 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00351 Means for dispensing and evacuation of reagents
• CPC Class Codes: B01J 2219/00367 capillary
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00527 Sheets
• 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 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00689 : using computers
• CPC Class Codes: B01J 2219/00702 : Processes involving means for analysing and characterising the products
• CPC Class Codes: B01J 2219/00713 : Electrochemical synthesis
• CPC Class Codes: B01J 2219/0072 : Organic compounds
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B01J 2219/00734 : Lipids
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12N 2533/52 : Fibronectin; Laminin
• CPC Class Codes: C12N 2535/10 : Patterned coating
• CPC Class Codes: C12Q 1/002 : Electrode membranes
• 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 60/14 : for creating libraries
• CPC Class Codes: G01N 2405/04 : Phospholipids, i.e. phosphoglycerides
• CPC Class Codes: G01N 33/5432 : Liposomes or microcapsules
• CPC Class Codes: G01N 33/54373 : involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/544 : the carrier being organic
• CPC Class Codes: G01N 33/6872 : Intracellular protein regulatory factors and their receptors, e.g. including ion channels
• CPC Class Codes: G01N 33/92 : involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis

Abstract

A surface detector array device suitable for use with a biosensor is disclosed. The device is formed of a substrate having a surface defining a plurality of distinct bilayer-compatible surface regions separated by one or more bilayer barrier regions. The bilayer-compatible surface regions carry on them, separated by a film of aqueous, supported fluid bilayers. The bilayers may contain selected receptors or biomolecules. A bulk aqueous phase covers the bilayers on the substrate surface. Multiplexed assays using the surface detector array device of the present invention are disclosed, as are automated methods for making the surface detector array device that enable formation of arrays wherein the composition of the individual, addressable bilayer regions is unrestricted.

Claims

1. A method for fabricating a surface detector array device, comprising the steps of:

providing a substrate having a surface defining a plurality of distinct bilayer-compatible surface regions separated by one or more bilayer barrier regions, wherein said bilayer-compatible surface regions and said bilayer barrier regions are formed of different materials;

providing a first suspension of lipid-bilayer vesicles having a first composition;

providing a second suspension of lipid-bilayer vesicles having a second composition;

transferring an aliquot of said first suspension to a first of said plurality of distinct bilayer-compatible surface regions;

transferring an aliquot of said second suspension to a second of said plurality of distinct bilayer-compatible surface regions;

incubating said substrate with said first and said second suspension to form a first lipid bilayer expanse stably localized above said first distinct bilayer-compatible surface region, and a second lipid bilayer expanse stably localized above said second distinct bilayer-compatible surface region, wherein each of said expanses is localized above each of said surface regions in the absence of covalent linkages between each of said lipid bilayer expanses and each of said bilayer-compatible surface regions, and is separated therefrom by an aqueous film interposed between said bilayer-compatible surface regions and said corresponding lipid bilayer expanses; and

establishing a bulk aqueous phase above said lipid bilayer expanses.

2. The method of claim 1, wherein said first composition differs from said second composition.

3. The method of claim 1, wherein said substrate comprises between about 10 and about 100 distinct bilayer-compatible surface regions.

4. The method of claim 3, wherein said substrate comprises at least about 2500 distinct bilayer-compatible surface regions.

5. The method of claim 4, wherein said substrate comprises at least about 25,000 distinct bilayer-compatible surface regions.

6. The method of claim 5, wherein said substrate comprises at least about 2.5 million distinct bilayer-compatible surface regions.

7. The method of claim 1, wherein the bilayer-compatible surface regions are separated from one another by bilayer barrier regions that are between about 1 μ

m and about 10 μ

m in width.

8. The method of claim 1, wherein said first composition further comprises a biomolecule.

9. The method of claim 8, wherein said biomolecule is selected from the group consisting of a transmembrane receptor and an ion channel.

10. The method of claim 9, wherein the biomolecule is covalently attached to a lipid molecule.

11. The method of claim 1, wherein said transferring steps are automated.

12. The method of claim 1, wherein said transferring steps comprise generating drops of said first and said second suspensions and touching said drops to said respective first and said second bilayer-compatible surface regions.

13. The method of claim 12, wherein each of said drops comprises less than 100 nl.

14. The method of claim 13, wherein each of said drops comprises less than 15 nl.

15. The method of claim 1, wherein said transferring steps comprise ejecting aliquots of said first and second suspensions across air spaces and onto said respective first and second bilayer-compatible surface regions.

16. The method of claim 15, wherein each of said ejected aliquots comprises less than 100 nl.

17. The method of claim 15, wherein each of said ejected aliquots comprises less than 15 nl.

18. An apparatus for fabricating a surface detector array device, comprising:

a platform for receiving a substrate, said substrate having a surface defining a plurality of distinct bilayer-compatible surface regions separated by one or more bilayer barrier regions, wherein said bilayer-compatible surface regions and said bilayer barrier regions are formed of different materials;

a source of a first suspension of lipid-containing vesicles or liposomes having a first composition;

a source of a second suspension of lipid-containing vesicles or liposomes having a second composition;

means for transferring an aliquot of said first suspension to a first of said plurality of distinct bilayer-compatible surface regions and for transferring an aliquot of said second suspension to a second of said plurality of distinct bilayer-compatible surface regions; and

a source of a bulk aqueous phase used to cover said fabricated surface detector array device.

19. The device of claim 18, further comprising a chamber for controlling one or more environmental parameters in the vicinity of the substrate surface.

20. The device of claim 19, wherein said parameters are selected from the group consisting of temperature, relative humidity, pressure, and illumination.

21. The device of claim 18, further comprising a microprocessor operatively programmed to control the relative movement between said stage and said transfer means.

34. The method of claim 18, wherein the bilayer-compatible surface regions are separated from one another by bilayer barrier regions that are between about 1 μ

m and about 10 μ

m in width.

22. A multiplexed assay, comprising the steps of:

providing a surface detection array device, said device comprising

a substrate having a surface defining a plurality of distinct bilayer-compatible surface regions separated by one or more bilayer barrier regions, said bilayer-compatible surface regions and said bilayer barrier regions being formed of different materials,

a first lipid bilayer expanse having a first composition and stably localized above a first of said plurality of distinct bilayer-compatible surface regions,

a second lipid bilayer expanse having a second composition different from said first composition and stably localized above a second of said plurality of distinct bilayer-compatible surface regions,

wherein each of said expanses is localized above each of said surface regions in the absence of covalent linkages between each of said lipid bilayer expanses and each of said bilayer-compatible surface regions, and is separated therefrom by an aqueous film interposed between said bilayer-compatible surface regions and said corresponding lipid bilayer expanses;

contacting said device with a bulk aqueous phase comprising a test agent; and

assaying an interaction between said test agent and said first composition and an interaction between said test agent and said second composition.

23. The method of claim 22, wherein said first composition comprises a first biomolecule, and said second composition comprises a second biomolecule.

24. The method of claim 23, wherein said biomolecules are selected from the group consisting of a transmembrane receptor and an ion channel.

25. The method of claim 23, wherein said first and said second biomolecules are different members of a receptor protein family.

26. The method of claim 22, wherein said first composition comprises a first lipid and said second composition comprises a second lipid.

27. The method of claim 22, wherein each of said first and said second lipid bilayer expanses comprises less than 5 μ

g.

28. The method of claim 27, wherein each of said first and said second lipid bilayer expanses comprises less than 1 μ

g.

29. The method of claim 28, wherein each of said first and said second lipid bilayer expanses comprises less than 0.5 μ

g.

30. The method of claim 22, wherein said substrate comprises between about 10 and about 100 distinct bilayer-compatible surface regions.

31. The method of claim 22, wherein said substrate comprises at least about 2500 distinct bilayer-compatible surface regions.

32. The method of claim 31, wherein said substrate comprises at least about 25,000 distinct bilayer-compatible surface regions.

33. The method of claim 32, wherein said substrate comprises at least about 2.5 million distinct bilayer-compatible surface regions.