Biosensors comprising a covalently attached monomolecular biological conjugate layer and a transducing device

• US 6,770,179 B1
• Filed: 03/02/1999
• Issued: 08/03/2004
• Est. Priority Date: 05/10/1993
• Status: Expired due to Fees

• Current Assignee: Universite De Montreal, L'
• Original Assignee: Universite De Montreal, L'
• Inventors: Nanci, Antonio
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US09/260,007
• Time in Patent Office: 1,981 Days
• Field of Search: 204/403, 435/817, 427/2.13
• US Class Current: 204/403.1
• CPC Class Codes: A61L 2300/252 Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
• CPC Class Codes: A61L 2300/254 Enzymes, proenzymes
• CPC Class Codes: A61L 2300/256 Antibodies, e.g. immunoglobulins, vaccines
• CPC Class Codes: A61L 2300/258 Genetic materials, DNA, RNA, genes, vectors, e.g. plasmids
• CPC Class Codes: A61L 2300/414 Growth factors
• CPC Class Codes: A61L 2300/64 Animal cells
• CPC Class Codes: A61L 27/28 Materials for coating prostheses
• CPC Class Codes: A61L 27/34 Macromolecular materials
• CPC Class Codes: A61L 27/54 Biologically active materials, e.g. therapeutic substances A61L27/227 takes precedence
• CPC Class Codes: B82Y 10/00 Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

The present invention relates to a biosensor which comprises a covalently attached monomolecular biological conjugate layer and a transducing system, wherein the biological conjugate layer has the following structural formula I:

—R—X—P  I

wherein,

R is O or S covalently attached via a first covalent bond to the transducing device surface;

X is a linker covalently attached to R via a second covalent bond and selected from a bond, linear or branched chains of 1 to 30 covalently attached atoms of at least C, N, O, Si or S, rings of at least one of C, N, O, Si or S, and a combination thereof; and

P is a biological molecule stably attached to X via a third covalent bond.

Claims

1. A biosensor comprising a covalently attached monomolecular biological conjugate layer and a transducing device, wherein the biological conjugate layer has the following structural formula I:

2. The biosensor of claim 1, wherein X is a bond, a linear alkyl C₁-C₃₀ chain, a linear chain consisting of from 1 to 20 C atoms interspersed with from 1 to 10 atoms of N, O or S, a ring composed of C and/or N, or a ring composed of C and/or N connected to a linear chain of C, N, O or S atoms, and X is terminated by a functional group which permits covalent linking to P.

3. The biosensor of claim 2, wherein the functional group is selected from the group consisting of COOH, NH₂, OH and SH.

4. The biosensor of claim 1, wherein said biological conjugate forms a self-assembling monomolecular layer.

5. The biosensor of claim 1, wherein the transducing device is a physiochemical transducer.

6. The biosensor of claim 1, wherein the transducing device is selected from the group consisting of optical transducers, electrochemical transducers, thermoelectric transducers, piezoelectric transducers, and magnetic transducers.

7. The biosensor of claim 1, wherein the transducing device is a microelectrode.

8. The biosensor of claim 7, further comprising a thin layer of metal coating the microelectrode, wherein said metal is selected from the group consisting of titanium, stainless steel, tantalum, Vitallium™

and an alloy thereof.

9. The biosensor of claim 1, wherein P is selected from the group consisting of biological tissue, microorganisms, whole cells, enzymes, receptors, antibodies and nucleic acids.

10. The biosensor of claim 1, wherein P is selected from the group consisting of osteopontin derivatized osteopontin, bone sialoprotein, bone acidic glycoprotein-75, osteocalcin, osteonectin, bone morphogenetic proteins, transforming growth factors, laminin, type IV collagen type VIII collagen, enamel proteins (amelogenins and non-amelogenins), α

₂HS-glycoprotein, fibronectin, cell adhesion peptides, prostaglandin, serum proteins, glucocorticosteroids (dexamethasone), phosphate, phosphoserine, pyrophosphates, phosphothreonine, phosvitin, phosphophoryn, biphosphonates, phosphonates, phosphatases, sulfonates, sulfates, carboxylates, bone and epithelial proteoglycans, mineral and cell binding peptide sequences such as Arginine-Glycine-Aspartic acid (Arg-Gly-Asp), polyaspartate, and other biological molecules capable of interacting with a biological moiety to be sensed or measured.