Bio-chip, photoluminescent methods for identifying biological material, and apparatuses for use with such methods and bio-chips

• US 6,706,479 B2
• Filed: 06/01/2001
• Issued: 03/16/2004
• Est. Priority Date: 10/05/2000
• Status: Expired due to Fees

• Current Assignee: Virginia Tech Intellectual Properties, Inc.
• Original Assignee: Virginia Tech Intellectual Properties, Inc.
• Inventors: Saraf, Ravi F., Niu, Sanjun
• Primary Examiner(s): Zitomer, Stephanie W.
• Assistant Examiner(s): Chakrabarti, Arun K.
• Application Number:
  US09/870,986
• Publication Number:
  US 20020042070A1
• Time in Patent Office: 1,019 Days
• Field of Search: 117/68, 250/214.1, 428/209, 435/6, 435/91.1
• US Class Current: 435/6.11
• CPC Class Codes: B01J 2219/00274 Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00497 Features relating to the solid phase supports
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00578 electrophoric
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• 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 2219/00637 by coating it with another layer
• 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 2219/00707 separated from the reactor apparatus
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00729 Peptide nucleic acids [PNA]
• CPC Class Codes: B82Y 30/00 : Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12Q 1/6825 : Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6837 : using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2523/319 : Photocleavage, photolysis, photoactivation
• 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: Y10T 428/24917 : including metal layer

Abstract

A method detects binding of molecules, advantageously without tagging molecules in the sample. A sensor is used in which is included a single stranded nucleic acid sequence and a photoluminescent material in respective layers. After the sensor is exposed to a biological sample for sufficient time for its single stranded nucleic acid sequence to bind to a material of interest, photoluminescence from the sensor can be measured. An apparatus for tagging-free detection of binding of molecules also is provided. Methods of making tagging-free sensors are provided. Also, tagging-free methods to detect binding of antigens and related devices are disclosed.

Claims

1. A tagging-free method to detect the binding of untagged single stranded nucleic acid to an untagged material of interest, comprising the steps of:

(A) providing a sensor comprised of a first layer and a second layer wherein said first layer comprises an untagged single stranded nucleic acid and wherein said second layer comprises a photoluminescent material, and wherein said first layer and said second layer are separate layers, and wherein said second layer comprises material selected from the group consisting of aromatic polymers, doped or undoped metal oxides, sulfides, arsenides, tellurides, and nitride and phosphide nanocomposites;

(B) exposing said sensor to a biological sample for sufficient time for said untagged single stranded nucleic acid to bind to an untagged material of interest in said biological sample;

(C) applying light to said sensor; and

(D) measuring photoluminescence from said sensor, wherein photoluminescence measured in said step of exposing is indicative of binding of said untagged single stranded nucleic acid to said untagged material of interest.

2. The tagging-free method of claim 1, wherein the single stranded nucleic acid is selected from the group consisting of DNA, RNA and PNA.

3. The tagging-free method of claim 1 wherein said second layer comprises a matrix material, with said photoluminescent material associated with said matrix material.

4. The method of claim 3 wherein said photoluminescent material is embedded in said matrix material.

5. The tagging-free method of claim 1, wherein the second layer comprises polystyrene.

6. The tagging-free method of claim 1, wherein the second layer comprises photoluminescent particles in a polymer matrix.

7. The tagging-free method of claim 6, wherein the photoluminescent particles are doped or undoped compounds selected from the group consisting of group II and group VI.

8. The tagging-free method of claim 7, including doped or undoped zinc sulfide.

9. The tagging-free method of claim 1, wherein the second layer comprises a nanocomposite.

10. The tagging-free method of claim 1, wherein said light is applied to said first layer of said sensor, and said light is ultraviolet light with wavelength in the range of 200-700 nm.

11. The tagging-free method of claim 10, wherein the wavelength of the ultraviolet light is in the range of 260-265 nm.

12. The tagging-free method of claim 1, wherein the first layer comprises an ssDNA monolayer.

13. The tagging-free method of claim 1, wherein the second layer comprises a thin-film or a support.

14. The tagging-free method of claim 1, wherein the second layer comprises a polymer.

15. The tagging-free method of claim 1, wherein the nucleic acid sequence is between 5 and 200 base pairs.

16. The tagging-free method of claim 15, wherein the sequence is about 25 base pairs.

17. The tagging-free method of claim 1, wherein the second layer has fluorescence when excited by light with a wavelength in the 200-700 nm range.

18. The tagging-free method of claim 1, wherein the sensor comprises ssDNA as said first layer grafted onto the second layer.

19. The tagging-free method of claim 1, wherein said first layer comprises a plurality of sections each of which comprises a different untagged single stranded nucleic acid sequence.

20. The method of claim 1, wherein said first layer is positioned on a first side of said second layer, and said measuring step measures photoluminescence from a second side of said second layer.

21. The method of claim 20, wherein said second side is opposite said first side on said second layer.

22. The method of claim 1, wherein said first layer is positioned on a first side of said second layer, and said measuring step measures photoluminescence reflected from said first side of said second layer.