Surface-bound, unimolecular, double-stranded DNA

• US 5,556,752 A
• Filed: 10/24/1994
• Issued: 09/17/1996
• Est. Priority Date: 10/24/1994
• Status: Expired due to Term

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Vetter, Dirk, Lockhart, David J., Diggelmann, Martin
• Primary Examiner(s): Fleisher, Mindy
• Assistant Examiner(s): Priebe, Scott David
• Application Number:
  US08/327,687
• Time in Patent Office: 694 Days
• Field of Search: 435/6, 536/23.1, 530/413
• US Class Current: 506/16
• CPC Class Codes: B01J 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• CPC Class Codes: B01J 2219/00529 DNA chips
• 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 2219/00619 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 2219/00722 Nucleotides
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• 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: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C12N 15/1093 General methods of preparing gene libraries, not provided for in other subgroups
• CPC Class Codes: C12Q 1/6811 Selection methods for production or design of target specific oligonucleotides or binding molecules
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2525/205 Aptamer
• CPC Class Codes: C12Q 2565/525 characterised by the capture oligonucleotide being double stranded
• CPC Class Codes: C40B 40/00 Libraries per se, e.g. arrays, mixtures
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: G01N 33/5308 : for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/551 : the carrier being inorganic

Abstract

Libraries of unimolecular, double-stranded oligonucleotides on a solid support. These libraries are useful in pharmaceutical discovery for the screening of numerous biological samples for specific interactions between the double-stranded oligonucleotides, and peptides, proteins, drugs and RNA. In a related aspect, the present invention provides libraries of conformationally restricted probes on a solid support. The probes are restricted in their movement and flexibility using double-stranded oligonucleotides as scaffolding. The probes are also useful in various screening procedures associated with drug discovery and diagnosis. The present invention further provides methods for the preparation and screening of the above libraries.

Claims

1. A synthetic unimolecular, double-stranded oligonucleotide library comprising a plurality of different members, each member having the formula:

space="preserve" listing-type="equation">Y--L.sup.1 --X.sup.1 --L.sup.2 --X.sup.2

wherein,

Y is a solid support;

X¹ and X² are a pair of complementary oligonucleotides;

L¹ is a spacer;

L² is a linking group having sufficient length such that X¹ and X² form a double-stranded oligonucleotide.

2. A library in accordance with claim 1, wherein L² is a polyethylene glycol group.

3. A library in accordance with claim 1, wherein X¹ and X² are complementary oligonucleotides each comprising of from 6 to 30 nucleic acid monomers.

4. A library in accordance with claim 1, wherein said solid support is a silica support and L¹ comprises an aminoalkylsilane and from 1 to 4 hexaethyleneglycols.

5. A library in accordance with claim 1, wherein said solid support is a silica support, L¹ comprises an aminoalkylsilane and from 1 to 4 hexaethyleneglycols, L² is a polyethyleneglycol group and X¹ and X² are complementary oligonucleotides each comprising of from 6 to 30 nucleic acid monomers.

6. A synthetic unimolecular, double-stranded oligonucleotide library of claim 1, wherein a portion of said double-stranded oligonucleotides formed by X¹ and X² further comprise a loop.