Spatially addressable, cleavable reflective signal elements, assay device and method

• US 6,312,901 B2
• Filed: 09/10/1999
• Issued: 11/06/2001
• Est. Priority Date: 07/08/1996
• Status: Expired due to Fees

• Current Assignee: Vindur Technologies, Inc.
• Original Assignee: Burstein Technologies, Inc.
• Inventors: Virtanen, Jorma
• Primary Examiner(s): Riley, Jezia
• Application Number:
  US09/394,137
• Publication Number:
  US 20010016316A1
• Time in Patent Office: 788 Days
• Field of Search: 435/6, 435/4, 435/5, 435/7.1, 435/7.21, 435/7.93, 435/7.94, 435/7.95, 435/810, 536/22.1, 436/501, 436/512
• 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 2219/00536 in the shape of disks
• CPC Class Codes: B01J 2219/0054 Means for coding or tagging the apparatus or the reagents
• 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 2219/00619 using hydrophilic or hydrophobic regions
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• 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: B01J 2219/00722 Nucleotides
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 70/00 Tags or labels specially adapted for combinatorial chemistry or libraries, e.g. fluorescent tags or bar codes
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54373 : involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/56988 : HIV or HTLV
• CPC Class Codes: G01N 33/5767 : non-A, non-B hepatitis
• CPC Class Codes: G01N 35/00069 : whereby the sample substrate is of the bio-disk type, i.e. having the format of an optical disk
• CPC Class Codes: Y10S 435/81 : Packaged device or kit

Abstract

A cleavable signal element for use in quantitative and qualitative assay devices and methods is described. Binding of the chosen analyte simultaneously to a first and a second analyte-specific side member of the cleavable signal element tethers the signal-responsive moiety to the signal element'"'"'s substrate-attaching end, despite subsequent cleavage at the cleavage site that lies intermediate the first and second side members. Assay devices comprising the cleavable signal elements are described, as are analytic methods adapted to their use. The analytic devices of the present invention may be adapted to detection using conventional CD-ROM and DVD readers.

Claims

1. A cleavable signal element, comprising:

a cleavable spacer, said cleavable spacer having a substratae-attaching end, a signal-responsive end, and a cleavage site intermediate said substrate-attaching end and said signal responsive end;

a signal responsive moiety;

a first side member adapted to bind a first site on a chosen analyte;

a second side member adapted to bind a second site of said chosen analyte;

said signal responsive moiety being attached to said cleavable spacer at said signal responsive end, said first side member being attached to said cleavable spacer intermediate said signal responsive end and said cleavage site, and said second side member being attached to said cleavable spacer intermediate said cleavage site and said substrate attaching end; and

said signal responsive moiety remaining bound to the substrate after cleavage at the cleavage site only when said first and second side members are bound to said chosen analyte.

2. The cleavable signal element of claim 1, wherein said signal responsive moiety is adapted to reflect or scatter incident light.

3. The cleavable signal element of claim 2, wherein said signal responsive moiety is a metal microsphere.

4. The cleavable signal element of claim 3, wherein said metal microsphere is essentially a metal selected from the group of gold, silver, nickel, platinum, chromium and copper.

5. The cleavable signal element of claim 4, wherein said metal microsphere is essentially of gold.

6. The cleavable signal element of claim 5, wherein said gold microsphere has a diameter between 1 nm-10 μ

m.

7. The cleavable signal element of claim 6, wherein said gold microsphere has a diameter between 0.5-5 μ

m.

8. The cleavable signal element of claim 7, wherein said gold microsphere has a diameter between 1-3 μ

m.

9. The cleavable signal element of claim 1, wherein said cleavage site is susceptible to chemical cleavage.

10. The cleavable signal element of claim 9, wherein said chemically susceptible cleavage site includes at least one siloxane group.

11. The cleavable signal element of claim 1, wherein said first side member and said second side member include oligonucleotides.

12. The cleavable signal element of claim 11, wherein said first and second side member oligonucleotides are 5 mers-20 mers.

13. The cleavable signal element of claim 12, wherein said first and second side member oligonucleotides are 8 mers-17 mers.

14. The cleavable signal element of claim 12, wherein said first and second side member oligonucleotides are 8 mers-12 mers.

15. The cleavable signal element of claim 1, wherein

said first side member includes a first member of a first specific binding pair,

said second side member includes a first member of a second specific binding pair, and

said second member of said first specific binding pair and said second member of said second specific binding pair are each present on the surface of a single analyte.

16. The cleavable signal element of claim 15, wherein said first member of said first specific binding pair includes a first antibody or antibody fragment and said first member of said second specific binding pair includes a second antibody, antibody fragment, or antibody derivative.

17. The cleavable signal element of claim 15, wherein

said first side member includes a first side member oligonucleotide,

said second side member includes a second side member oligonucleotide,

said first member of said first specific binding pair includes a first binding pair oligonucleotide,

said first member of said second specific binding pair includes a second binding pair oligonucleotide, and

said first side member oligonucleotide includes sequence complementary to sequence included in said first binding pair oligonucleotide, said second side member oligonucleotide includes sequence complementary to sequence included in said second binding pair oligonucleotide, and said complementary sequences are noncovalently associated.