Nanoparticles biosensor

• US 5,922,537 A
• Filed: 11/08/1996
• Issued: 07/13/1999
• Est. Priority Date: 11/08/1996
• Status: Expired due to Fees

• Current Assignee: CareStream Health Incorporated
• Original Assignee: No Slashed AB Immunoassay Incorporated
• Inventors: Ewart, Thomas G., Bogle, Gavin T.
• Primary Examiner(s): Zitomer, Stephanie W.
• Application Number:
  US08/746,420
• Time in Patent Office: 977 Days
• Field of Search: 435/6, 435/5, 435/7.1, 435/7.92, 435/7.93, 422/68.1, 436/501, 436/536, 436/518, 436/524, 436/527, 436/532, 436/526, 436/525
• US Class Current: 435/6.11
• 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: C12Q 1/6823 Release of bound markers
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/587 Nanoparticles
• CPC Class Codes: G01N 33/588 with semiconductor nanocrystal label, e.g. quantum dots

Abstract

Biosensor technology based on the labelling entities having particle reporters provides cost competitive readily manufactured assay devices. Sub-micron particles of uniform dimension in metals, polymers, glasses, ceramics and biological structures such as phages are used as the labelling entities. Such reporter particles greatly increase the sensitivity and accuracy, and provide a variety of assay techniques for determining analyte presence in a sample. The particles may have dielectric, paramagnetic and/or phosphorescent properties, such particles are particularly useful in a variety of competition type assays. Novel phosphor and phage particles are provided for use as unique labelling entities.

Claims

1. A displacement competition assay for determining sample analyte concentration in a sample fluid, comprising:

(i) applying said sample fluid to a test surface having a binding agent bound through specific recognition sites to an immobilized analyte, wherein said immobilized analyte is affixed to a solid support and said binding agent is labelled with phages encapsulating reporter particles, said reporter particles providing a base level signal; and

(ii) binding said sample analyte to at least one of said specific recognition sites wherein binding of said sample analyte to said specific recognition site displaces said labelled binding agent away from said test surface into said sample fluid to define a test level signal, wherein displacement of said labelled binding agent is related to the concentration of sample analyte in said sample fluid;

(iii) removing said sample fluid from said test surface; and

(iv) comparing said base level signal to said test level signal to determine the concentration of said sample analyte.

2. The competition assay of claim 1, wherein said labelled binding agent and said sample analyte form a complex and wherein said complex is displaced into said sample fluid.

3. The competition assay of claim 1, wherein said test surface is provided on one or both of two parallel plates spaced apart at a distance so as to effect capillary movement of said sample fluid across said test surface.

4. The competition assay of claim 1, wherein said reporter particles are paramagnetic, said test level signal being measured in inductance when said test surface is exposed to a magnetic field.

5. The competition assay of claim 1, wherein said reporter particles are dielectric, said test level signal being measured in capacitance when said test surface forms part of a capacitor.

6. The competition assay of claim 5, wherein said test surface comprises a ferroelectric layer.

7. The competition assay of claim 6, wherein said ferroelectric layer comprises barium titanate.

8. The competition assay of claim 1, wherein said reporter particles have at least one of a dielectric, magnetic or phosphorescent property.

9. The competition assay of claim 1, wherein said reporter particles are phosphorescent, said test level signal being detected as phosphorescent radiation when said test surface is stimulated to cause said particles to phosphoresce.

10. The competition assay of claim 1, wherein said reporter particles are individually in the range of 1 to 1000 nanometers.

11. The competition assay of claim 1, wherein said reporter particles are individually in the range of 1 to 1000 microns.

12. The competition assay of claim 1, wherein said phages encapsulating reporter particles express on their surface said binding agent and are reversibly bound through specific recognition sites to said immobilized analyte, and wherein said sample analyte competes with said binding agent for recognition sites on said immobilized analyte and displaces said phage particle, wherein displacement of said phages encapsulating reporter particles defines a test level signal.

13. The competition assay of claim 1, wherein said phages encapsulating reporter particles express on their surface said binding agent and bind reversibly to specific recognition sites on said immobilized analyte, and wherein said sample analyte competes with said immobilized analyte for recognition sites on said binding agent and forms a complex with said binding agent to displace said phage particle from said test surface, wherein said phages encapsulating reporter particles and sample analyte complex defines a test level signal.