Porous substrates for DNA arrays

• US 6,994,972 B2
• Filed: 03/18/2002
• Issued: 02/07/2006
• Est. Priority Date: 09/02/1999
• Status: Expired due to Fees

• Current Assignee: Corning Incorporated
• Original Assignee: Corning Incorporated
• Inventors: Lahiri, Joydeep, Bookbinder, Dana C., Tepesch, Patrick D., Bardhan, Pronob, Wusirika, Raja R., Tanner, Cameron W.
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US10/101,144
• Publication Number:
  US 20020142339A1
• Time in Patent Office: 1,422 Days
• Field of Search: 435/6, 435/7.1, 435/174, 435/283.1, 435/287.2, 422/68.1, 536/23.1
• US Class Current: 435/6.11
• CPC Class Codes: B01J 20/28033 Membrane, sheet, cloth, pad, lamellar or mat
• CPC Class Codes: B01J 20/28057 Surface area, e.g. B.E.T specific surface area
• CPC Class Codes: B01J 20/28083 being in the range 2-50 nm, i.e. mesopores
• CPC Class Codes: B01J 20/28097 being coated, filled or plugged with specific compounds
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00533 essentially rectangular
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/0063 Other, e.g. van der Waals forces, hydrogen bonding
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00641 the porous medium being continuous, e.g. porous oxide substrates
• 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 2219/00722 Nucleotides
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C03C 11/005 obtained by leaching after a phase separation step
• CPC Class Codes: C03C 17/30 with silicon-containing compounds
• CPC Class Codes: C03C 2218/111 by dipping, immersion
• CPC Class Codes: C40B 40/06 Libraries containing nucleotides or polynucleotides, or derivatives thereof

Abstract

A planar, rigid substrate made from a porous, inorganic material coated with cationic polymer molecules for attachment of an array of biomolecules, such as DNA, RNA, oligonucleotides, peptides, and proteins. The substrate has a top surface with about at least 200 to about 200,000 times greater surface area than that of a comparable, non-porous substrate. The cationic polymer molecules are anchored on the top surface and in the pores of the porous material. In high-density applications, an array of polynucleotides of a known, predetermined sequence is attached to this cationic polymer layer, such that each of the polynucleotide is attached to a different localized area on the top surface. The top surface has a surface area for attaching biomolecules of approximately 387,500 cm²/cm² of area (˜7.5 million cm²/1×3 inch piece of substrate). Each pore of the plurality of pores in the top surface of the substrate has a pore radius of between about 40 Å to about 75 Å. Not only does the cationic coating in and over the pores of the substrate greatly increase the overall positive charge on the substrate surface, but also given the size of the pores provides binding sites to which biomolecules can better attach.

Claims

1. A method for preparing a planar, rigid porous glass substrate for attachment of biomolecules, the method comprising:

treating a porous glass substrate with a cationic polymer wherein the step of treating said porous glass substrate further comprises;

coating said porous glass substrate by applying a layer of cationic polymer, the cationic polymer having at least one amino group and dissolved in a buffer at a pH above the pKa of the amino group;

washing said porous glass substrate such that said cationic polymer electrostatically interacts with pores and surface of said porous glass substrate;

rinsing said porous glass substrate with water; and

drying said porous glass substrate.

2. The method according to claim 1 wherein drying comprises the step of heating said porous glass substrate to about 140–

1750 C to cross-link free silanol groups.

3. The method according to claim 1, wherein said pores of said porous glass substrate ranges in size of between 40–

200 |.

4. The method according to claim 2, wherein the temperature to which said porous glass substrate is heated is about 150–

170°

C.

5. The method according to claim 1, wherein said cationic polymer is applied by means of dip-coating.

6. The method according to claim 1, wherein said cationic polymer includes at least one of the following:

polylysine, polyethylene-imine, aminosilanes, cationic dendrimers, star polymers, and polyvinylamine.

7. The method according to claim 1, wherein said cationic polymer is either polylysine or polyethylene-imine.

8. The method according to claim 1, wherein said cationic polymer is y-amino-propyltriethoxysilane.

9. The method according to claim 1, further comprising the step of attaching biomolecules to said porous glass substrate, said biomolecules are selected from the group consisting of:

DNA, RNA, oligonucleotides, peptides, and proteins.