Triggered optical biosensor

• US 6,297,059 B1
• Filed: 06/22/1999
• Issued: 10/02/2001
• Est. Priority Date: 06/22/1998
• Status: Expired due to Fees

• Current Assignee: Los Alamos National Security LLC (Government of the United States of America)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Song, Xuedong, Swanson, Basil I.
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): PHAM, MINH QUAN K
• Application Number:
  US09/338,457
• Time in Patent Office: 833 Days
• Field of Search: 436/501, 436/518, 436/514, 436/2, 436/172, 435/4, 435/7.1, 435/287.1, 435/287.2, 435/288.7
• US Class Current: 436/501
• CPC Class Codes: G01N 2405/04 Phospholipids, i.e. phosphoglycerides
• CPC Class Codes: G01N 2405/10 Glycosphingolipids, e.g. cerebrosides, gangliosides
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/5436 with ligand physically entrapped within the solid phase liposomes G01N33/5432; immunological test elements G01N33/54386
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

An optical biosensor is provided for the detection of a multivalent target biomolecule, the biosensor including a substrate having a bilayer membrane thereon, a recognition molecule situated at the surface, the recognition molecule capable of binding with the multivalent target biomolecule, the recognition molecule further characterized as including a fluorescence label thereon and as being movable at the surface and a device for measuring a fluorescence change in response to binding between the recognition molecule and the multivalent target biomolecule.

Claims

1. An optical biosensor for detection of a multivalent target biomolecule comprising:

a substrate having a fluid membrane thereon;

recognition molecules situated at a surface of said fluid membrane, said recognition molecule capable of binding with said multivalent target biomolecule and said recognition molecule linked to a single fluorescence molecule and as being movable upon said surface of said fluid membrane; and

,

a means for measuring a change in fluorescent properties in response to binding between multiple recognition molecules and said multivalent target biomolecule.

2. A method of detecting a multivalent target biomolecule comprising:

contacting a sample with a sensor including a substrate having a fluid membrane thereon, recognition molecules situated at a surface of said fluid membrane, said recognition molecule capable of binding with said multivalent target biomolecule and said recognition molecule linked to a single fluorescence molecule and as being movable upon said surface of said fluid membrane; and

measuring a change in fluorescent properties in response to binding between multiple recognition molecules and said multivalent target biomolecule.

3. An optical biosensor for detection of a multivalent target biomolecule comprising:

a substrate having a fluid membrane thereon;

at least two different recognition molecules situated movably upon a surface of said fluid membrane, said recognition molecules capable of binding with said multivalent target biomolecule wherein at least one of said recognition molecules is linked to a single fluorescence donor molecule and at least another of said recognition molecules is linked to a single fluorescence acceptor molecule; and

,

a means for measuring a change in fluorescent properties in response to binding between said two different recognition molecules and said multivalent target biomolecule.

4. The optical biosensor of claim 3 wherein the recognition molecule is contained within an upper layer of a bilayer surface.

5. A method of detecting a multivalent target biomolecule comprising:

contacting a sample with a sensor including a substrate having a fluid membrane thereon and at least two different recognition molecules situated movably upon a surface of said fluid membrane, said recognition molecules capable of multivalent binding with said multivalent target biomolecule wherein at least one of said recognition molecules is linked to a single fluorescence donor molecule and at least another of said recognition molecules is linked to a single fluorescence acceptor molecule; and

measuring a change in fluorescence in response to binding between said two different recognition molecules and said multivalent target biomolecule.

6. An optical biosensor for detection of a multivalent target biomolecule comprising:

a recognition molecule capable of binding with said multivalent target biomolecule wherein said recognition molecule is linked to a single fluorescence molecule selected from the group consisting of a fluorescence donor molecule and a fluorescence acceptor molecule thereon;

a supporting surface for said recognition molecule, wherein said supporting surface includes a fluorescence label selected from the group consisting of a fluorescence donor label and a fluorescence acceptor label thereon wherein said supporting surface including the fluorescence donor label when said recognition molecule includes the single fluorescence acceptor molecule and said supporting surface including the fluorescence acceptor label when said recognition molecule includes the single fluorescence donor molecule; and

,

a means for measuring a change in fluorescent properties in response to binding between multiple recognition molecules and said multivalent target biomolecule.

7. The optical biosensor of claim 6 wherein said recognition molecule is labeled with a single fluorescence donor molecule and said supporting surface is a phospholipid bilayer labeled with a fluorescence acceptor label.

8. The optical biosensor of claim 7 wherein said multivalent target biomolecule is cholera, said recognition molecule is ganglioside GM1 labeled with a singe fluorescence donor molecule selected from the group consisting of DABCY and BODIPY-FL and said phospholipid bilayer supporting surface is beta-palmitoyl-gamma-oleoyl-L-alpha-phosphatidylcholine labeled with a fluorescence acceptor label of pyrene.

9.The optical biosensor of claim 6 wherein the supporting surface is a phospholipid bilayer and the recognition molecule is contained within an upper layer of the bilayer surface.

9. The optical biosensor of claim 6 wherein the fluorescence label upon the recognition molecule has the fluorescent properties of self-quenching.

10. An optical biosensor for the detection of a target biomolecule comprising:

a substrate having a fluid membrane thereon;

recognition molecules situated at a surface of said fluid membrane, said recognition molecule bound with a selected multivalent biomolecule, said recognition molecules, each linked to a single fluorescence molecule; and

,

a means for measuring a change in fluorescent properties in response to competitive binding with said selected multivalent biomolecule by said recognition molecules and said target biomolecule.