Up-converting reporters for biological and other assays using laser excitation techniques

• US 6,399,397 B1
• Filed: 10/18/2000
• Issued: 06/04/2002
• Est. Priority Date: 09/14/1992
• Status: Expired due to Fees

• Current Assignee: SRI International, Inc.
• Original Assignee: SRI International, Inc.
• Inventors: Dyer, Mark J., Ng, Steve Y., Schneider, Luke V., Zarling, David A., Rossi, Michel J., Peppers, Norman A., Kane, James, Faris, Gregory W.
• Primary Examiner(s): Chin, Christopher L.
• Assistant Examiner(s): DO, PENSEE T
• Application Number:
  US09/690,823
• Time in Patent Office: 594 Days
• Field of Search: 436/518, 436/528, 436/540, 436/546, 436/164, 436/169, 436/172, 436/177, 436/800, 436/805, 435/970, 435/973, 435/7.1, 435/7.5
• US Class Current: 436/518
• CPC Class Codes: B01J 2219/00315 Microtiter plates
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00707 separated from the reactor apparatus
• 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 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6818 involving interaction of two or more labels, e.g. resonant energy transfer
• CPC Class Codes: C12Q 2537/143 Multiplexing, i.e. use of multiple primers or probes in a single reaction, usually for simultaneously analyse of multiple analysis
• CPC Class Codes: C12Q 2563/00 Nucleic acid detection characterized by the use of physical, structural and functional properties
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 33/533 : with fluorescent label
• CPC Class Codes: G01N 33/582 : with fluorescent label
• CPC Class Codes: G01N 33/588 : with semiconductor nanocrystal label, e.g. quantum dots
• CPC Class Codes: Y10S 435/97 : Test strip or test slide
• CPC Class Codes: Y10S 435/973 : Simultaneous determination of more than one analyte
• CPC Class Codes: Y10S 436/80 : Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]

Abstract

The invention provides methods, compositions, and apparatus for performing sensitive detection of analytes, such as biological macromolecules and other analytes, by labeling a probe molecule with an up-converting label. The up-converting label absorbs radiation from an illumination source and emits radiation at one or more higher frequencies, providing enhanced signal-to-noise ratio and the essential elimination of background sample autofluorescence. The methods, compositions, and apparatus are suitable for the sensitive detection of multiple analytes and for various clinical and environmental sampling techniques.

Claims

1. An apparatus for determining the presence of at least one target analyte in a liquid sample comprising:

a lateral surface for receiving a liquid sample and transporting the liquid sample to a capture area on the lateral surface, wherein a portion of the lateral surface contains a mobilizable labeled probe comprised of an up-converting phosphor microparticle label attached to a probe which binds specifically to the target analyte to form a labeled probe-analyte complex; and

wherein the capture area has immobilized thereon a capture probe capable of binding specifically to the labeled probe-analyte complex.

2. The apparatus of claim 1, wherein the lateral surface is comprised of a chromatographic material.

3. The apparatus of claim 2, wherein the lateral surface is a wick.

4. The apparatus of claim 1, wherein the probes are antibodies or antibody fragments and the analyte is an antigen.

5. The apparatus of claim 1, wherein the capture probe is biotinylated and the up-converting phosphor label is bound to streptavidin or avidin.

6. The apparatus of claim 1, wherein the lateral surface contains at least two distinct mobilizable labeled probes capable of binding specifically to at least two distinct target analytes to form distinct labeled probe-analyte complexes, and wherein the up-converting phosphor labels differ in their absorption and/or emission spectra.

7. The apparatus of claim 1, wherein the target analyte is selected from the group consisting of polynucleotides, polypeptides, viruses, microorganisms, haptens, mammalian cells, steroid hormones, glycoproteins, lipoproteins, biotinylated magnetic beads, prescribed or over-the-counter drugs, illegal substances, intoxicants and drugs of abuse.

8. The apparatus of claim 1, wherein the probes are selected from the group consisting of antibodies, polynucleotides, polypeptide hormones, avidin, streptavidin, Staphylococcus aureus Protein A. lectins, antigens and mixtures thereof.

9. The apparatus of claim 1, wherein the target analyte is a polynucleotide and the probes are polynucleotides which specifically hybridize to the target polynucleotide.

10. The apparatus of claim 1, wherein the up-converting phosphor microparticle label is smaller than about 3 microns in diameter.

11. The apparatus of claim 10, wherein the up-converting phosphor microparticle label is smaller than about 1 micron in diameter.

12. The apparatus of claim 11, wherein the up-converting phosphor microparticle label is about 0.1 to about 0.5 microns in diameter.

13. The apparatus of claim 1, wherein the up-converting phosphor microparticle comprises a phosphor host material doped with at least one activator couple comprised of an absorber ion and an emitter ion.

14. The apparatus of claim 13, wherein the absorber ion is ytterbium and the emitter ion is selected from the group consisting of erbium, holmium, thulium, and terbium.

15. The apparatus of claim 14, wherein the phosphor host material is selected from the group consisting of oxysulfides, oxyhalides, fluorides, gallates, silicates, aluminates, phosphates, and vanadates.

16. The apparatus of claim 15, wherein the phosphor host material is selected from the group consisting of sodium yttrium fluoride, lanthanum fluoride, lanthanum oxysulfide, yttrium oxysulfide, yttrium fluoride, yttrium gallate, yttrium aluminum garnet, gadolinium fluoride, barium yttrium fluoride, and gadolinium oxysulfide.

17. The apparatus of claims 16, wherein the up-converting phosphor microparticle comprises sodium yttrium fluoride ytterbium erbium or yttrium ytterbium erbium oxysulfide.

18. In an apparatus for determining the presence of at least one target analyte in a liquid sample comprising:

a lateral surface for receiving a liquid sample and transporting the liquid sample to a capture area on the lateral surface, wherein a portion of the lateral surface contains a mobilizable labeled probe which binds specifically to the target analyte to form a labeled probe-analyte complex; and

wherein the capture area has immobilized thereon a capture probe capable of binding specifically to the labeled probe-analyte complex,

the improvement characterized in that the label is an up-converting phosphor label.