Up-converting reporters for biological and other assays

• US 6,312,914 B1
• Filed: 10/18/2000
• Issued: 11/06/2001
• Est. Priority Date: 09/14/1992
• Status: Expired due to Term

• Current Assignee: Orasure Technologies Incorporated
• Original Assignee: Orasure Technologies Incorporated
• Inventors: Faris, Gregory W., Dyer, Mark J., 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
• Primary Examiner(s): Zitomer, Stephanie W.
• Assistant Examiner(s): Wilder, Cynthia B.
• Application Number:
  US09/690,828
• Time in Patent Office: 384 Days
• Field of Search: 435/6, 435/5.29, 435/7.1, 435/189, 435/325, 536/23.1, 536/24.3, 530/387.1, 530/350, 250/484.2, 250/484.3, 250/909, 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 photophysical catalysis method which induces mutagenesis or cytotoxicity in a cell or causes photolytic decomposition of a chemical present in the cell environment, comprising the steps of:

(a) contacting a cell with a pair of matched up-converting and luminescent labeled probes under conditions sufficient to bind the up-converting labeled probe and the luminescent labeled probe to the cell to form a labeled probe-cell complex, wherein the up-converting label is an inorganic phosphor particle comprising at least one rare earth element and a phosphor host material; and

wherein the up-converting label or the luminescent label is an excitation label and the other is an emission label; and

,

(b) illuminating the labeled probe-cell complex to excite the excitation label which in turn excites the emission label such that the emission label emits radiation at a wavelength which is mutagenic or cytotoxic to the cell or is of a wavelength that causes photolytic decomposition of a chemical present in the cell environment.

2. A photophysical catalysis method which induces cytotoxicity in a cell, comprising the steps of:

(a) contacting a cell with a pair of matched up-converting and dye sensitizer labeled probes under conditions sufficient to bind the up-converting labeled probe and the dye sensitizer labeled probe to the cell to form a labeled probe-cell complex, wherein the up-converting label is an inorganic phosphor particle comprising at least one rare earth element and a phosphor host material; and

,

(b) illuminating the labeled probe-cell complex to excite the up-converting label which in turn excites the dye sensitizer such that the dye sensitizer is excited to a triplet state to induce cytotoxic activity in the cell.

3. A method according to claim 2, wherein the excited sensitizer dye transfers its energy to a dissolved molecular oxygen molecule to yield an excited oxygen molecule.

4. A photophysical catalysis method which induces mutagenesis or cytotoxicity in a cell or causes photolytic decomposition of a chemical present in the cell environment, comprising the steps of:

(a) contacting a cell with an up-converting probe under conditions sufficient to bind the up-converting labeled probe to the cell to form a labeled probe-cell complex, wherein the up-converting label is an inorganic phosphor particle comprising at least one rare earth element and a phosphor host material; and

(b) illuminating the labeled probe-cell complex such that the label emits radiation at a wavelength which is mutagenic or cytotoxic to the cell or is of a wavelength that causes photolytic decomposition of a chemical present in the cell environment.