Treatment of substrates for immobilizing biomolecules

• US 6,787,312 B2
• Filed: 08/09/2001
• Issued: 09/07/2004
• Est. Priority Date: 08/09/2001
• Status: Expired due to Fees

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: He, Mingqian, Li, Guangshan, Raghavachari, Nalini, Xie, Xinying, Bao, Yijia, Xie, Yuming
• Primary Examiner(s): WHISENANT, ETHAN C
• Application Number:
  US09/925,808
• Publication Number:
  US 20030036062A1
• Time in Patent Office: 1,125 Days
• Field of Search: 435/6, 536/23.1, 536/24.3
• US Class Current: 506/39
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Abstract

A method of treating a substrate for immobilizing a biomolecule and substrates produced by the method are disclosed. The method includes contacting at least a portion of a substrate with a reducing agent such as a hydride. Treatment with an appropriate reducing agent substantially eliminates autofluorescence on substrates.

Claims

1. A method for reducing auto-fluorescence on a substrate, the method comprising:

providing a substrate with at least a first surface, said first surface being either with or without either a biological or a synthetic molecule immobilized thereon, or having either a coated or uncoated surface with a residual fluorescence; and

treating at least a portion of the first surface of the substrate with a reducing agent for longer than about 5 minutes to reduce auto-fluorescence on the substrate, wherein said autofluorescence is reduced by at least an order of magnitude RFU.

2. The method of claim 1, wherein the reducing agent includes hydrogen.

3. The method of claim 1, wherein the reducing agent includes a hydride.

4. The method of claim 3, wherein the reducing agent includes a borohydride.

5. The method of claim 1, wherein the step of treating the substrate with a reducing agent includes contacting at least a portion of the first surface of the substrate with an aqueous solution containing between 0.1 and 1% sodium borohydride by volume.

7. The method of claim 1, wherein said substrate is treated for at least 10 minutes with said reducing agent.

8. The method of claim 1, further comprising a step of scanning the substrate.

9. The method of claim 8, wherein said canning step includes scanning the substrate for a fluorescent label.

10. The method of claim 1, wherein said substrate is made from a material selected from the group consisting of inorganic materials, glass, ceramic materials, metals, and semiconductor materials.

11. The method of claim 1, wherein said substrate is made from a material selected from the group consisting of organic materials, polyesters, polybutylene terephthalate, polyvinylchloride, polyvinylidene fluoride, polytetrafluoroethylene, polycarbonate, polyamide, poly(meth)acrylate, polystryrene, polyethylene or theylene/vinyl acetate copolymer.

12. The method of claim 1, wherein said biological or synthetic molecule includes at least one of the following species:

ribonucleic acids (RNA), deoxyribonucleic acids (DNA), synthetic oligonucleotides, antibodies, proteins, peptides, lectins, modified polysaccharides, cells, synthetic composite macromolecules, functionalized nanostructures, synthetic polymers, modified/blocked nucleotides/nucleosides, modified/blocked amino acids, fluorophores, chromophores, ligands, chelates, and haptens.

13. A substrate having an array of biomolecules non-covalently attached thereto produced by the method of claim 1.

14. The substrate of claim 1, wherein the biomolecules are nucleic acids or oligonucleotides.

15. The substrate of claim 1, wherein the substrate is contains an array of nucleic acids or oligonucleotides.

6. The method of 5 wherein the aqueous solution contains between 0.2% and 0.3% sodium borohydride by volume.