Biomolecules having multiple attachment moieties for binding to a substrate surface

• US 7,833,715 B1
• Filed: 01/18/2007
• Issued: 11/16/2010
• Est. Priority Date: 01/11/2000
• Status: Expired due to Fees

• Current Assignee: Sanofi-Aventis SA (Sanofi )
• Original Assignee: Nanogen Recognomics GmbH
• Inventors: Windhab, Norbert, Schweitzer, Markus, Greef, Charles, Havens, John R., Wang, Daguang, Onofrey, Thomas J.
• Primary Examiner(s): Riley; Jezia
• Application Number:
  US11/655,009
• Publication Number:
  US 20100286377A1
• Time in Patent Office: 1,398 Days
• Field of Search: 435/6, 536/23.1, 536/24.3, 536/25.3
• US Class Current: 435/6.1
• CPC Class Codes: C07H 19/04 Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
• 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 1/68 involving nucleic acids
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393

Abstract

Methods of binding biomolecules to a substrate are provided that include contacting the biomolecule with a branched linking moiety to form a branched linking structure. The branched linking structure is then contacted with a binding moiety on the substrate to form a coupled substrate binding structure, thereby binding the biomolecule to the substrate. The biomolecule may contain a Lewis base or a nucleophile to react with a Lewis acid or electrophile in the branched linking moiety. Alternatively, the biomolecule may contain a Lewis acid or electrophile that can react with a Lewis base or nucleophile in the branched linking moiety. Additionally, the biomolecule can be bound to the substrate through a covalent or non-covalent bond.

Claims

1. A method of binding a biomolecule to a substrate comprising the steps of:

providing a biomolecule;

contacting the biomolecule with a branched linking moiety to form a branched linking structure that is adapted to couple with the substrate, wherein the branched linking structure has multiple reactive groups; and

contacting the branched linking structure with the substrate to form a coupled substrate binding structure, where the biomolecule is bound to the substrate through multiple linkages.

2. The method of claim 1, further comprising the step of activating the branched linking structure before contacting the branched linking structure with the binding moiety.

3. The method of claim 1, wherein the substrate is a polymer.

4. The method of claim 1, wherein the substrate is an activated polymer.

5. The method of claim 1, wherein the substrate is in contact with an electronically addressable microchip.

6. The method of claim 1, wherein the substrate is an activated glass slide.

7. The method of claim 1, wherein the substrate is a glass slide.

8. The method of claim 1, wherein the biomolecule comprises a Lewis base or a nucleophile.

9. The method of claim 1, wherein the biomolecule comprises a Lewis acid or an electrophile.

10. The method of claim 8, wherein the Lewis base or the nucleophile is selected from the group consisting of alcohol, amine, hydrazine, hydrazide, salicylic hydroxamic acid, and sulfhydryl.

11. The method of claim 9, wherein the Lewis acid or the electrophile is selected from the group consisting of an epoxide, aziridine, vinyl, aldehyde, ketone, acetal, disulfide, carboxylic acid, amide, bromo or iodoacetamide, N-hydroxysuccinimidyl ester, sulfo-N-hydroxysuccinimidyl ester, azlactone, isocyanate, thioisocyanate, phenyl boronic acid, and carbonate.

12. The method of claim 1, wherein the branched linking moiety comprises a phosphoramidite.

13. The method of claim 12, wherein the phosphoramidite is selected from the group consisting of Diethyl 3-[(2-cyanoethoxy)(diisopropylamino)phosphanyloxy]glutarate;

Diethyl 5-{[(2-cyanoethoxy)(diisopropylamino)phosphanyloxy]methyl}isophthalate;

Dimethyl 3,3′

-(2-{[(2-cyanoethoxy)(diisopropylamino)phosphanyloxy]methyl}-2-{[2-(methoxycarbonyl)ethoxy]methyl}propane-1,3-diylbisoxy)dipropionate;

Ethyl 6-[(2-cyanoethoxy)(diisopropylamino)phosphanyloxy]hexanoate; and

6-[(2-Cyanoethoxy)(diisopropylamino)phosphanyloxy)-N′

-tritylhexanohydrazide.

14. The method of claim 1, wherein multiple linkages are covalent bonds.

15. The method of claim 1, wherein the multiple linkages are non-covalent bonds.