Up-converting reporters for biological and other assays

• US 6,159,686 A
• Filed: 01/30/1998
• Issued: 12/12/2000
• Est. Priority Date: 09/14/1992
• Status: Expired due to Term

• Current Assignee: SRI International, Inc., STC Technologies Corporation
• Original Assignee: SRI International, Inc.
• Inventors: Kardos, Keith W., Ng, Steve Y., Niedbala, R. Sam, Schneider, Luke V., Burton, Jarrett Lee, Cooper, David E., Zarling, David A., Rossi, Michel J., Peppers, Norman A., Kane, James, Faris, Gregory W., Dyer, Mark J.
• Primary Examiner(s): Zitomer, Stephanie W.
• Assistant Examiner(s): WILDER, CYNTHIA B
• Application Number:
  US09/016,402
• Time in Patent Office: 1,047 Days
• Field of Search: 435/5, 435/6, 435/4, 435/7.1, 436/501, 436/518, 436/800, 536/23.1, 536/24.3, 530/350, 530/387.1, 250/909, 250/484.3, 250/484.2, 216/25
• US Class Current: 435/6.14
• 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 2219/00637 by coating it with another layer
• 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 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• 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 2021/6441 : with two or more labels
• 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 21/645 : Specially adapted constructive features of fluorimeters
• CPC Class Codes: G01N 21/6452 : Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/6454 : using an integrated detector array
• CPC Class Codes: G01N 33/533 : with fluorescent label
• CPC Class Codes: G01N 33/542 : with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: G01N 33/582 : with fluorescent label
• CPC Class Codes: G01N 33/588 : with semiconductor nanocrystal label, e.g. quantum dots

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. A composition for detecting an analyte in a sample using matched labels, comprising:

a pair of matched up-converting and luminescent labeled probes, wherein the up-converting labeled probe is an inorganic phosphor particle comprising at least one rare earth element and a phosphor host material, and the luminescent label is matched to the up-converting label.

2. The composition according to claim 1, wherein the probes are antibodies, polynucleotides, polypeptide hormones, receptor ligands, streptavidins, Staphylococcus aureus Protein A, lectins, antigens or mixtures thereof.

3. The composition according to claim 1, wherein the target analyte is a polynucleotide, a polypeptide, a virus, a microorganism, a hapten, a mammalian cell, a hormone, a glycoprotein, a blood clotting factor, a lipoprotein, a drug, a polysaccharide, a metabolite, a pollutant, a pesticide or mixtures thereof.

4. The composition according to claim 1, wherein the composition is in the form of an assay kit.

5. The composition according to claim 1, wherein the up-converting label is an up-converting inorganic phosphor and comprises at least one rare earth element and the probes are antibodies, polynucleotides, polypeptide hormones, receptor ligands, streptavidins, Staphylococcus aureus Protein A, lectins, antigens or mixtures thereof.

6. A method for detecting an analyte in a sample with the composition of claim 1 using matched labels, comprising the steps of:

contacting a sample suspected of containing a target analyte with matched up-converting and luminescent labeled probe pairs, wherein the up-converting labeled probe and the luminescent labeled probes are capable of binding the target analyte to form a labeled probe-target complex;

illuminating the sample to excite a label of the labeled probe-target complex; and

observing the sample for emission radiation from the matched label of the labeled probe-target complex, thereby detecting the presence or absence of the analyte.

7. A method according to claim 6, wherein the up-converting label is an excitation label and the luminescent label is an emission label.

8. A method according to claim 7, wherein the illumination step excites the excitation label with radiation having a wavelength ranging from about 700 to about 1500 nm.

9. A method according to claim 6, wherein the luminescent label is an excitation label and the up-converting label is an emission label.

10. A method according to claim 9, wherein the illumination step excites the excitation label with radiation having a wavelength ranging from about 350 to about 1500 nm.

11. A method according to claim 10, wherein the illumination step excites the excitation label with radiation having a wavelength ranging from about 350 to about 700 nm.

12. A method according to claim 6, wherein the sample is contacted with two or more matched labeled probe pairs, the matched labeled probe pairs having matched labels with different excitation label wavelengths and different emission label wavelengths, and wherein the matched probe pairs bind to different target analytes.

13. A method according to claim 6, wherein the sample is contacted with two or more matched labeled probe pairs, the matched labeled probe pairs having matched labels with similar excitation label wavelengths and different emission label wavelengths, and wherein the matched probe pairs bind to different target analytes.

14. A method according to claim 6, wherein the sample is contacted with two or more matched labeled probe pairs, the matched labeled probe pairs having matched labels with different excitation label wavelengths and similar emission label wavelengths, and wherein the matched probe pairs bind to different target analytes.

15. A method according to claim 6, wherein the sample is contacted with two or more matched labeled probe pairs, the matched labeled probe pairs having matched labels with similar excitation and emission label wavelengths, and wherein the matched labels are attached to probes which bind affinitively to different target analytes.

16. A method according to claim 6, wherein the target analyte is a polynucleotide, a polypeptide, a virus, a microorganism, a hapten, a mammalian cell, a hormone, a glycoprotein, a blood clotting factor, a lipoprotein, a drug, a polysaccharide, a metabolite, a pollutant, a pesticide and mixtures thereof.

17. A method according to claim 6, wherein the probes are antibodies, polynucleotides, polypeptide hormones, receptor ligands, streptavidins, Staphylococcus aureus Protein A, lectins, antigens and mixtures thereof.

18. A method according to claim 6, wherein one of the labeled probes is bound to a solid support.

19. A method according to claim 6, further comprising before the illumination step, the step of separating the labeled-probe target complex from any unbound labeled probes.