US5674698A — Up-converting reporters for biological and other …

Claim 1

Independent Claim 124 words

A method for detecting an analyte in a sample, comprising the steps of: contacting a sample containing a target analyte with a labeled binding component to specifically bind the target analyte and form a labeled binding component-target complex, wherein the labeled binding component comprises a binding component attached to an up-converting inorganic phosphor particle comprising at least one rare earth element and a phosphor host material and being capable of converting excitation radiation to emission radiation of a shorter wavelength; separating any unbound labeled binding component from the labeled binding component-target complex; illuminating the labeled binding component-target complex with excitation radiation; and detecting emission radiation of at least one label emission wavelength, wherein the emission radiation has a shorter wavelength than the excitation radiation.

Claim 2

Independent Claim 124 words

A method for detecting an analyte in a sample, comprising the steps of: contacting a sample containing a target analyte with a labeled binding component to specifically bind the target analyte and form a labeled binding component-target complex, wherein the labeled binding component comprises a binding component attached to an up-converting inorganic phosphor particle comprising at least one rare earth element and a phosphor host material and being capable of converting excitation radiation to emission radiation of a shorter wavelength; separating any unbound labeled binding component from the labeled binding component-target complex; illuminating the labeled binding component-target complex with excitation radiation; and detecting emission radiation of at least one label emission wavelength, wherein the emission radiation has a shorter wavelength than the excitation radiation.

Claim 3

Dependent Claim 20 words

A method according to claim 1, wherein said up-converting inorganic phosphor comprises ytterbium and erbium in a phosphor host material.

Claim 4

Dependent Claim 22 words

A method according to claim 2, wherein the up-converting inorganic phosphor comprises sodium yttrium fluoride ytterbium erbium or yttrium ytterbium erbium oxysulfide.

Claim 5

Dependent Claim 30 words

A method of claim 1, further comprising, before the contacting step, the step of attaching the up-converting inorganic phosphor particle to the binding component to form the labeled binding component.

Claim 6

Dependent Claim 20 words

A method according to claim 4, wherein the binding component is attached to the label by covalent or noncovalent binding.

Claim 7

Dependent Claim 23 words

A method according to claim 5, wherein the binding component is streptavidin or avidin and the target analyte is a biotinylated target analyte.

Claim 8

Dependent Claim 24 words

A method of claim 1, wherein the labeled binding component-target complex is separated from the unbound labeled probe by immobilization on a solid support.

Claim 9

Dependent Claim 43 words

A method according to claim 7, wherein the step of separating unbound labeled binding component from the labeled binding component-target complex in the sample is performed by washing the sample with an aqueous solution to remove suspendible or soluble unbound labeled binding component.

Claim 10

Dependent Claim 28 words

A method of claim 7, wherein the labeled binding component target complex is bound to a first binding component on the solid support to form a sandwich complex.

Claim 11

Dependent Claim 42 words

A method according to 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.

Claim 12

Dependent Claim 26 words

A method according to claim 1, wherein the step of illuminating with a label excitation wavelength is performed with an infrared laser diode or light-emitting diode.

Claim 13

Dependent Claim 17 words

A method according to claim 1, wherein the infrared laser diode or light-emitting diode emits pulsed illumination.

Claim 14

Dependent Claim 20 words

A method according to claim 12, wherein the infrared laser diode or light-emitting diode is pulsed through direct current modulation.

Claim 15

Dependent Claim 27 words

A method according to claim 12, wherein the step of detecting light emission of at least one label emission wavelength is performed by time-gated or lock-in detection.

Claim 16

Dependent Claim 18 words

A method according to claim 11, wherein the step of detecting light emission is performed with phase-sensitive detection.

Claim 17

Dependent Claim 27 words

A method according to claim 11, wherein the laser diode or light-emitting diode has peak emissions in the range of 960-980 nm and at approximately 1500 nm.

Claim 18

Dependent Claim 31 words

A method according to claim 1, wherein said step of detecting light emission is performed with a photomultiplier, photodiode, a charge coupled device, a charge injection device, or photographic film emulsion.

Claim 19

Dependent Claim 21 words

A method According to claim 1, wherein the target analyte is immobilized in a histological tissue section or a solid support.

Claim 20

Dependent Claim 29 words

A method according to claim 1, wherein the binding component is selected from the group consisting of: antibodies, polynucleotides, polypeptide hormones, streptavidin, Staphylococcus aureus Protein A, lectins, and antigens.

Claim 21

Independent Claim 127 words

A method for detecting an analyte in a sample, comprising the steps of: contacting a sample containing a target analyte with a binding component to specifically bind the target analyte and form a binding component-target complex; contacting the binding component-target complex with a label to form a labeled binding component-target complex, the label comprising an up-converting inorganic phosphor particle comprising at least one rare earth element and a phosphor host material and being capable of converting excitation radiation to emission radiation of a shorter wavelength; separating any unbound label from the labeled binding component-target complex; illuminating the labeled binding component-target complex with excitation radiation; and detecting emission radiation of at least one label emission wavelength, wherein the emission radiation has a shorter wavelength than the excitation radiation.

Claim 22

Independent Claim 127 words

A method for detecting an analyte in a sample, comprising the steps of: contacting a sample containing a target analyte with a binding component to specifically bind the target analyte and form a binding component-target complex; contacting the binding component-target complex with a label to form a labeled binding component-target complex, the label comprising an up-converting inorganic phosphor particle comprising at least one rare earth element and a phosphor host material and being capable of converting excitation radiation to emission radiation of a shorter wavelength; separating any unbound label from the labeled binding component-target complex; illuminating the labeled binding component-target complex with excitation radiation; and detecting emission radiation of at least one label emission wavelength, wherein the emission radiation has a shorter wavelength than the excitation radiation.

Claim 23

Dependent Claim 29 words

A method according to claim 20, wherein the binding component is a primary antibody and the up-converting inorganic phosphor is bound to the binding component through a secondary antibody.

Claim 24

Dependent Claim 21 words

A method according to claim 21, wherein said secondary antibody is biotinylated and said up-converting inorganic phosphor is bound to streptavidin.

Claim 25

Dependent Claim 30 words

A method according to claim 20, wherein the target analyte is a polynucleotide and the binding component is a biotinylated polynucleotide which hybridizes to the target polynucleotide under binding conditions.

Claim 26

Dependent Claim 18 words

A method according to claim 23, wherein the up-converting inorganic phosphor comprises an up-converting phosphor particle and streptavidin.

Claim 27

Dependent Claim 30 words

A method of claim 20 further comprising, before the contacting step, the step of attaching the up-converting inorganic phosphor particle to the binding component to form the labeled binding component.

Claim 28

Independent Claim 126 words

A method for detecting a biotinylated analyte in a sample, comprising the steps of: contacting a sample containing a biotinylated analyte with a labelled binding component to specifically bind the biotinylated analyte and form a labeled binding component-target complex, wherein the labeled binding component comprises a streptavidin-coated up-converting inorganic phosphor particle, the up-converting inorganic phosphor particle comprising at least one rare earth element and a phosphor host material and being capable of converting excitation radiation to emission radiation of a shorter wavelength; separating any unbound labeled binding component from the labeled binding component-target complex; illuminating the labeled binding component-target complex with excitation radiation; and detecting emission radiation of at least one label emission wavelength, wherein the emission radiation has a shorter wavelength than the excitation radiation.

Claim 29

Independent Claim 126 words

A method for detecting a biotinylated analyte in a sample, comprising the steps of: contacting a sample containing a biotinylated analyte with a labelled binding component to specifically bind the biotinylated analyte and form a labeled binding component-target complex, wherein the labeled binding component comprises a streptavidin-coated up-converting inorganic phosphor particle, the up-converting inorganic phosphor particle comprising at least one rare earth element and a phosphor host material and being capable of converting excitation radiation to emission radiation of a shorter wavelength; separating any unbound labeled binding component from the labeled binding component-target complex; illuminating the labeled binding component-target complex with excitation radiation; and detecting emission radiation of at least one label emission wavelength, wherein the emission radiation has a shorter wavelength than the excitation radiation.

Claim 30

Dependent Claim 14 words

A method according to claim 26, wherein the target is a biotinylated magnetic bead.

Claim 31

Dependent Claim 30 words

A method of claim 26 further comprising, before the contacting step, the step of attaching the up-converting inorganic phosphor particle to the binding component to form the labeled binding component.

Claim 32

Independent Claim 133 words

A method for detecting an analyte in a sample, comprising the steps of: contacting a sample containing a target analyte with a labeled binding component to specifically bind the target analyte and form a labeled binding component-target complex, wherein the labeled binding component comprises a binding component attached to an up-converting inorganic phosphor particle comprising at least one rare earth element and a phosphor host material and being capable of converting excitation radiation to emission radiation of a shorter wavelength; differentiating the labeled binding component-target complex from any unbound labeled binding component in the sample; illuminating the labeled binding component-target complex with excitation radiation; and detecting emission radiation of at least one label emission wavelength from the labeled binding component-target complex, wherein the emission radiation has a shorter wavelength than the excitation radiation.

Claim 33

Independent Claim 133 words

A method for detecting an analyte in a sample, comprising the steps of: contacting a sample containing a target analyte with a labeled binding component to specifically bind the target analyte and form a labeled binding component-target complex, wherein the labeled binding component comprises a binding component attached to an up-converting inorganic phosphor particle comprising at least one rare earth element and a phosphor host material and being capable of converting excitation radiation to emission radiation of a shorter wavelength; differentiating the labeled binding component-target complex from any unbound labeled binding component in the sample; illuminating the labeled binding component-target complex with excitation radiation; and detecting emission radiation of at least one label emission wavelength from the labeled binding component-target complex, wherein the emission radiation has a shorter wavelength than the excitation radiation.

Claim 34

Dependent Claim 35 words

The method of claim 29, wherein the labeled binding component-target complex is illuminated with a confocal beam having a focal point at the contact surface and being divergent at points other than the contact surface.

Claim 35

Dependent Claim 30 words

A method of claim 29, further comprising, before the contacting step, the step of attaching the up-converting inorganic phosphor particle to the binding component to form the labeled binding component.

Claim 36

Dependent Claim 58 words

A method of claim 29, wherein the differentiating step comprises contacting the labeled binding component-target complex with a contact surface wherein the labeled binding component-target complex is localized at the contact surface as compared to the unbound labeled binding component; and the illuminating step comprises illuminating the labeled binding component-target complex at the contact surface with excitation radiation.

Claim 37

Dependent Claim 109 words

The method of claim 32, wherein binding component-target complexes are localized to the contact surface by a method selected from the group consisting of: magnetic localization of magnetic beads to said contact surface, wherein said binding component-target complexes are localized on the magnetic beads relative to unbound labeled binding component; gravitational sedimentation of binding component-target complexes from unbound labelled binding component, wherein said sedimented binding component-target complexes are localized on the contact surface relative to unbound labeled binding component; filtration over a contact surface wherein said binding component-target complexes are localized on the contact surface relative to unbound labeled binding component; antibody capture; affinity adsorption; and nucleic acid hybridization.

Claim 38

Dependent Claim 16 words

A method of claim 29, wherein the differentiating and illuminating steps are accomplishied by confocal excitation.

Claim 39

Dependent Claim 20 words

A method of claim 29 wherein the differentiating, illuminating, and detecting steps are accomplished by confocal excitation and confocal detection.

Claim 40

Dependent Claim 14 words

A method of claim 29, wherein the differentiating step is accomplished by size discrimination.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	2000-10-06	2001-04-07	2001-10-07	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	2004-10-06	2005-04-07	2005-10-07	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2008-10-06	2009-04-07	2009-10-07	$7700.00	Window Closed / Lapsed

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

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