Target molecule attachment to surfaces

• US 6,465,178 B2
• Filed: 01/08/1999
• Issued: 10/15/2002
• Est. Priority Date: 09/30/1997
• Status: Expired due to Term

• Current Assignee: Surmodics Incorporated
• Original Assignee: Surmodics Incorporated
• Inventors: Chappa, Ralph A., Swan, Dale G., Swanson, Melvin J., Guire, Patrick E., Hu, Sheau-Ping
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): FORMAN, BETTY J
• Application Number:
  US09/227,913
• Publication Number:
  US 20010014448A1
• Time in Patent Office: 1,376 Days
• Field of Search: 435/6, 435/174, 435/287.2, 435/287.9, 422/68.1, 536/23.1, 536/24.3
• US Class Current: 435/6.12
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00531 essentially square
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• 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 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00716 Heat activated synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• CPC Class Codes: C07C 235/84 with the carbon atom of at least one of the carboxamide groups bound to a carbon atom of a six-membered aromatic ring
• CPC Class Codes: C07C 323/42 Y being a carbon atom of a six-membered aromatic ring
• CPC Class Codes: C07C 45/63 by introduction of halogen; by substitution of halogen atoms by other halogen atoms
• CPC Class Codes: C07C 49/813 : polycyclic
• CPC Class Codes: C07H 21/00 : Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• 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 60/14 : for creating libraries

Abstract

Method and reagent composition for covalent attachment of target molecules, such as nucleic acids, onto the surface of a substrate. The reagent composition includes groups capable of covalently binding to the target molecule. Optionally, the composition can contain photoreactive groups for use in attaching the reagent composition to the surface. The reagent composition can be used to provide activated slides for use in preparing microarrays of nucleic acids.

Claims

1. An activated slide comprising a support surface coated with a bound composition of a reagent composition, the bound composition comprising a polymeric backbone having more than one pendent thermochemically amine-reactive or sulfhydryl-reactive groups configured and arranged to form covalent bonds with fundamental groups on a target molecule, the reagent composition being coated and immobilized on the surface in a manner that permits:

a) small sample volume of a solution containing the target molecule to be applied in the form of a discrete spot on the reagent composition-coated surface, b) target molecule present in the sample volume to become attached to the bound composition by reaction between the functional groups of the target molecule and the thermochemically amine-reactive or sulfhydryl-reactive groups of the bound composition, and c) substantially all unattached target molecule to be washed from the sport without undue detectable amounts of target molecule in the area surrounding the spot, and wherein the bound composition is configured and arranged to form covalent bonds with functional groups on the target molecule without the use of attracting groups to attract the target molecule to the bound composition.

2. An activated slide according to claim 1 wherein the activated slide is adapted for fabricating a microarray wherein the target molecule comprises a nucleic acid and the surface comprises the surface of a plastic, silicon hydride, or organosilane-pretreated glass or silicone slide.

3. An activated slide according to claim 2 wherein the nucleic acid comprises one or more functional groups selected from the group consisting of amine and sulfhydryl groups.

4. An activated slide according to claim 1 wherein the activated slide is configured and arranged to receive a sample in an amount of twenty nanoliters or less.

5. An activated slide according to claim 1 wherein the reagent composition comprises one or more latent reactive groups comprising photoreactive groups for attaching the reagent composition to the surface of the slide upon application of energy from a suitable source, wherein the latent reactive groups immobilize the composition onto the slide.

6. An activated slide according to claim 1 wherein the reagent composition comprises thermochemically reactive groups and photoreactive groups, wherein the thermochemically reactive groups and the photoreactive groups are pendent from the polymeric backbone and the photoreactive groups are selected from the group consisting of photoreactive aryl ketones.

7. An activated slide according to claim 6 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and heterocyclic analogs of anthrone.

8. An activated slide according to claim 1 wherein the polymeric backbone is selected from the group consisting of acrylics, vinyls, nylons, polyurethanes and polyethers, the pendent thermochemically reactive groups are selected from the group consisting of activated esters, epoxides, azlactones, activated hydroxyls, aldehydes, isocyanates, thioisocyanates, carboxylic acid chlorides, alkyl halides, maleimide, and α

-iodoacetamide, and the backbone further comprises one or more pendent photoreactive groups selected from the group consisting of aryl ketones.

9. An activated slide for binding target molecule in a sample, the slide comprising:

a. a support surface; and

b. a bound composition attached to the support surface,

wherein the bound composition comprises a polymeric backbone having more than one thermochemically amine-reactive or sulfhydryl-reactive groups attache thereto, and wherein the bound composition is configured and arranged to form covalent bonds with functional groups on the target molecule without use of attracting groups to attract the target molecule to the bound composition.

10. An activated slide according to claim 9 wherein the activated slide is adapted for fabricating a microarray wherein the target molecule comprises a nucleic acid and the surface comprises the surface of a plastic, silicon hydride, or organosilane-pretreated glass or silicone slide.

11. An activated slide according to claim 10 wherein the nucleic acid comprises one or more functional groups selected from the group consisting of amine and sulfhydryl groups.

12. An activated slide according to claim 9 wherein the activated slide is configured and arranged to receive a sample in an amount of twenty nanoliters or less.

13. An activated slide according to claim 9 wherein the bound composition comprises a reaction product of a reagent composition with the surface, wherein the reagent composition comprises one or more photoreactive groups for attaching the reagent composition to the surface of the slide upon application of energy from a suitable source, wherein the photoreactive groups immobilize the reagent composition onto the slide to form the bound composition.

14. An activated slide according to claim 13 wherein the reagent composition comprises thermochemically reactive groups and photoreactive groups, wherein the thermochemically reactive and photoreactive groups are pendent from one or more hydrophilic polymeric backbones and the photoreactive groups are photoreactive aryl ketones.

15. An activated slide according to claim 14 wherein the photoreactive aryl ketones are each, independently, selected from the group consisting of acetophenone, benzophenone, anthraquinone, anthrone, and heterocyclic analogs of anthrone.

16. An activated slide according to claim 9 wherein the polymeric backbone is selected from the group consisting acrylics, vinyls, nylons, polyurethanes and polyethers, the pendent thermochemically reactive groups are selected from the group consisting of activated esters, epoxides, azlactones, activated hydroxyls, aldehydes, isocyanates, thioisocyanates, carboxylic acid chlorides, alkyl halides, maleimide, and α

-iodoacetamide, and the backbone further comprises one or more pendent photoreactive groups selected from the group consisting of aryl ketones.