FLUORESCENT LABELING REAGENT

• US 20080124806A1
• Filed: 02/26/2007
• Published: 05/29/2008
• Est. Priority Date: 02/27/2006
• Status: Active Grant

• Current Assignee: KEIO University
• Original Assignee: KEIO University
• Inventors: Isobe, Tetsuhiko, Ohkubo, Michio, Aizawa, Hideki, Saito, Daisuke, Noda, Kunihiro, Takagi, Tomohiro, Asakura, Ryo
• Granted Patent: US 7,842,505 B2
• Time in Patent Office: Days
• US Class Current: 436/56
• 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: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex
• CPC Class Codes: G01N 33/588 with semiconductor nanocrystal label, e.g. quantum dots
• CPC Class Codes: Y10T 436/13 Tracers or tags

Abstract

A fluorescent labeling reagent of the present invention includes an inorganic fluorescent particle and a material (A) having a material (B) of biological origin adsorbed or bound thereto. The inorganic fluorescent particle is integrated with the material (A) so as to form the reagent of the present invention. The inorganic fluorescent particle used in the present invention is capable of emitting light with a wavelength of 650 nm to 1600 nm in the infrared region or the near-infrared region which can be detected by means of Si-CCD or InGaAs-PD and can penetrate an H₂O rich sample when excited by light with a wavelength of 650 nm or longer which has the shortest transparent wavelength of AlInGaP-LD including oxygen adsorption type hemoglobin used for DVDs etc.

Claims

1. A fluorescent labeling reagent comprising:

an inorganic fluorescent particle capable of emitting light with a wavelength of 650 nm to 1600 nm when excited by light with a wavelength of 650 nm or longer; and

a material (A) having a material (B) of biological origin adsorbed or bound thereto, the material (A) being integrated with the inorganic fluorescent particle.

2. The fluorescent labeling reagent according to claim 1,

wherein the inorganic fluorescent particle has a primary particle size of 1 nm to 5000 nm.

3. The fluorescent labeling reagent according to claim 1, wherein the inorganic fluorescent particle comprises a rare earth element as a dopant having a fluorescent spectrum in near-infrared region.

4. The fluorescent labeling reagent according to claim 1,

wherein the inorganic fluorescent particle comprises one or more rare earth element as dopants selected from the group consisting of cerium, praseodymium, neodymium, gadolinium, holmium, erbium, thulium, ytterbium, europium, terbium, samarium, and dysprosium.

5. The fluorescent labeling reagent according to claim 1,

wherein the inorganic fluorescent particle has a garnet structure of yttrium, aluminum and oxygen and is a crystalline particle in which at least one of rare earth elements capable of producing fluorescence is incorporated into the garnet structure.

7. The fluorescent labeling material reagent according to claim 1,

wherein the material (A) is a functional bead of an inorganic material or polymer material of which surface is chemically modified to have a molecule capable of adsorbing or binding to a specific biomolecule, and

the integration of the inorganic fluorescent material and the material (A) is carried out by dispersing the inorganic fluorescent particle into the functional bead.

10. The fluorescent labeling reagent according to claim 7,

wherein the functional bead has a diameter of 0.1 μ

m to 100 μ

m and is selected from the group consisting of polymer beads including polystyrene bead, polypropylene bead, cross-linked acrylic bead, and polylactide bead, magnetic beads, glass beads, metallic beads, and the surface of the functional bead is chemically modified to specifically adsorb or bind to the material of biological origin.

6. (canceled)

8-9. -9. (canceled)

11. A fluorescent labeling reagent comprising:

an inorganic fluorescent particle capable of emitting light with a wavelength of 650 nm to 1600 nm when excited by light with a wavelength of 650 nm or longer; and

a material of biological origin adsorbed or bound to the inorganic fluorescent particle.

12. The fluorescent labeling reagent according to claim 11,

wherein the inorganic fluorescent particle has a primary particle size of 1 nm to 5000 nm.

13. The fluorescent labeling reagent according to claim 11, wherein the inorganic fluorescent particle comprises a rare earth element as a dopant having a fluorescent spectrum in near-infrared region.

14. The fluorescent labeling reagent according to claim 11,

wherein the inorganic fluorescent particle comprises one or more rare earth element as dopants selected from the group consisting of cerium, praseodymium, neodymium, gadolinium, holmium, erbium, thulium, ytterbium, europium, terbium, samarium, and dysprosium.

15. The fluorescent labeling reagent according to claim 11,

wherein the inorganic fluorescent particle has a garnet structure of yttrium, aluminum and oxygen and is a crystalline particle in which at least one of earth elements capable of producing fluorescence is incorporated into the garnet structure.

16. The fluorescent labeling reagent according to claim 11,

wherein the material of the biological origin is adsorbed or bound to the inorganic fluorescent particle by means of chemical reaction in which a modification group introduced onto a surface of the inorganic fluorescent particle is directly bound to an organic molecule of the material of the biological origin.

17. The fluorescent labeling reagent according to claim 16, wherein the direct binding of the inorganic fluorescent particle and the material (B) is carried out by the steps of:

subjecting the inorganic fluorescent particle to a surface treatment by means of reaction with silane coupling agent having amino group, thiol group, or carboxyl group so that the amino group, thiol group, or carboxyl group is introduced onto a surface of the inorganic fluorescent particle, and

binding the amino group, thiol group, or carboxyl group to the material (B) selected from the group consisting of avidin, streptavidin, fusion protein of avidin or streptavidin, biotin, antigen, antibody, DNA, and RNA.

18. The fluorescent labeling reagent according to claim 17,

wherein the introduction of amino group is carried out by using aminoalkyl thiol compound and utilizing a binding of the inorganic fluorescent particle and thiol.