WO2002056020A1 - Complexes d'un solide et de polymeres utilises dans un dosage biologique - Google Patents

Complexes d'un solide et de polymeres utilises dans un dosage biologique Download PDF

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Publication number
WO2002056020A1
WO2002056020A1 PCT/JP2002/000121 JP0200121W WO02056020A1 WO 2002056020 A1 WO2002056020 A1 WO 2002056020A1 JP 0200121 W JP0200121 W JP 0200121W WO 02056020 A1 WO02056020 A1 WO 02056020A1
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Prior art keywords
group
polymer
complex
pair
semiconductor
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PCT/JP2002/000121
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English (en)
Japanese (ja)
Inventor
Kazunori Kataoka
Yukio Nagasaki
Hidenori Otsuka
Takehiko Ishii
Yuka Sunaga
Original Assignee
Kazunori Kataoka
Yukio Nagasaki
Hidenori Otsuka
Takehiko Ishii
Yuka Sunaga
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Application filed by Kazunori Kataoka, Yukio Nagasaki, Hidenori Otsuka, Takehiko Ishii, Yuka Sunaga filed Critical Kazunori Kataoka
Priority to JP2002556224A priority Critical patent/JPWO2002056020A1/ja
Publication of WO2002056020A1 publication Critical patent/WO2002056020A1/fr

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
    • G01N33/588—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with semiconductor nanocrystal label, e.g. quantum dots
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase

Definitions

  • the present invention relates to a conjugate for incorporation into a biological assay system and to a preparation comprising the conjugate.
  • a wide variety of detection means have been provided as a method for detecting an analyte present in a biological sample.
  • One widely used technique is to use colorimetry, including light absorption analysis.
  • colorimetry including light absorption analysis.
  • a method for detecting a polynucleotide based on a gold nanoparticle probe modified with a mercaptoalkyloligonucleotide has been reported (R. Elghanian et al., SCIENCE III Vol. 277). 1997, pp. 1078-1081). It is taught that the method using this probe can detect oligonucleotides with high selectivity and high sensitivity.
  • the present inventors have proposed that a certain functional group such as a mercapto group or a functional group such as a Selective and high-sensitivity measurement of analytes, with the composite of a polymer having a polymer segment and a specific metal being hardly adversely affected by contaminants other than the analyte in the biological sample Found that it can be used to Moreover, when the metal is fine particles having a submicron size (less than about 100 nm), it was confirmed that the complex was stable in an aqueous solution so that it could be handled by capillary electrophoresis. did it.
  • the semiconductor nanocrystals (or particles) having a surface layer made of the polymer serve as donors for fluorescence resonance energy transfer in a detection system of an analyte. Found that it can be used.
  • a preparation comprising a complex of a solid substance and a polymer for constructing a biological analyte detection system utilizing a change in light absorption wavelength or a fluorescence emission wavelength. Things are provided. And this preparation is
  • the analyte can be one of the members forming a specific binding pair non-covalently,
  • the solid substance is selected from a substance group consisting of a metal and a semiconductor,
  • the polymer is based on a polyethylene oxide (PEG) segment and is a member of a molecule capable of forming a specific binding pair with an analyte (one of the members) at one end of the molecule.
  • the complex is formed through a bond between a functional group having an unshared electron pair and the surface of the substance,
  • an organism having a first antibody or a first complementary chain to an analyte (for example, antigen or DNA) at one end of a polymer molecule in the complex, and using a complex in which a solid substance is a semiconductor.
  • analyte for example, antigen or DNA
  • a method for detecting a body is also provided.
  • a second antibody or a second complementary chain to the analyte, which is modified with a certain fluorescent dye is allowed to coexist for detection.
  • PEG having hydrophilicity and high mobility on a metal or semiconductor surface is assumed to have a plurality of brush-like structures. Etc. can be avoided.
  • each analyte is specifically adsorbed by binding between members forming a specific binding pair.
  • the binding of the PEG brush-like structure to the surface is extremely stable in an aqueous solution, and does not substantially cleave even after, for example, capillary electrophoresis.
  • FIG. 1 is a conceptual diagram of a method for multiplex detection of an analyte DNA using a preparation of a semiconductor complex according to the present invention.
  • R, G, and B in the figure indicate that they are colored red, green, and blue, respectively, and those with an apostrophe attached to those symbols mean that the color tone has changed from the corresponding one. I do.
  • FIG. 2 is an electropherogram showing the results of capillary electrophoresis of the CdS complex obtained in Example 1.
  • FIG. 3 is an electropherogram showing the result of capillary electrophoresis of the Au complex obtained in Example 2.
  • FIG. 4 shows a composite particle (semiconductor C d S particle having a biotinylated PEG on its surface: a structure capable of forming a specific binding pair) according to the present invention described in Example 3.
  • the member is piotin.
  • FIG. 7 is a diagram showing the change in the intensity of the absorption maximum of a mixture of adenosine and avidin modified with the fluorescent dye Texas Red. It has been shown that the transfer of fluorescent energy from a semiconductor to a fluorescent dye occurs in a concentration-dependent manner.
  • FIG. 5 is a graph showing the relationship between the complex particle concentration and the emission intensity of avidin modified with Texas Red.
  • the solid substance forming the complex according to the present invention can be in any shape, but is preferably intended to be a plate, a strip, a microchip and a microparticle having a size of a submicron. I have. Further, from the viewpoint of utilizing the stability of the complex in an aqueous solvent, it is preferable that the complex is in the form of fine particles that can incorporate capillary electrophoresis into a biological analyte detection system.
  • sub-micron size usually means that the average diameter of the particles is less than 100 m, but for the purpose of the present invention it is not necessarily less than 100 zm However, the term is used to mean particles of extremely small size.
  • a size capable of performing electrophoresis with a capillary having an inner diameter of 25 to 100 ⁇ m is understood as the size of the fine particles of the present invention. ing. More specifically, the average particle size is from 0.1 nm to several microns, preferably from 1 nm to 100 nm, particularly preferably 10 ⁇ ! 1100 nm.
  • Commercially available microparticles can be used to prepare composites of solid materials of this size, (metal or semiconductor) microparticles, or, for example, the formation of colloidal precipitates of metals or semiconductors known per se Method, desalination or reduction treatment of each salt solution O
  • any metal can be used regardless of its type, as long as the change in plasmon absorption associated with the change in surface shape (for example, thickening) due to the binding of the analyte to the complex can be detected. it can. However, preferred are gold, silver, platinum, and aluminum.
  • the semiconductor may be any type of semiconductor as long as it emits fluorescent light. Examples thereof include I n P and I n belonging to the III-V type (or the 3B group-5B group). Preferred are As and GaAs, and CdS, CdSe, ZeSe and ZeS belonging to the type II-VI (or 2B-1B). Can be mentioned.
  • the solid substance as described above is provided as a complex with a polymer.
  • Polymers for the purposes of the present invention are based (or main) on the PEG segment.
  • the PEG segment On the basis of that the PEG segment is in a state where it imparts sufficient hydrophilicity to the surface of the solid substance and acts to avoid non-specific adsorption of contaminants other than the analyte in the biological sample.
  • Means From another point of view having one or more repeating units of ethylene oxide in one molecule of the polymer is preferably 3 or more, more preferably 5 or more, and particularly preferably 10 or more, based on the PEG segment.
  • the polymer is covalently bound to one member residue (to the analyte) or one of the other members (to the analyte) that can form a specific binding pair with a given analyte at one end of the molecule.
  • Having a functional group capable of forming This functional group can be covalently bound to the other member of the analyte, one of the members that can form a specific binding pair non-covalently (eg, hydrogen bond, hydrophobic bond, etc.). It is.
  • poly It is intended for (or oligo) nucleotides, proteins, sugars, hormones, etc., and as a combination of members capable of forming a specific binding pair, a pair of an antigen and an antibody, a pair of a biotin and avidin, and a sugar And lectins; hormones or signal transducers and corresponding receptor proteins; enzymes and their substrates or inhibitors; and nucleic acid fragments consisting of certain nucleotide sequences and stringent conditions. It is intended for use with pairs of nucleic acid fragments that form a hybrid below. Any of the members of these pairs may be the subject.
  • the biological sample containing such a subject includes biological fluids (urine, blood, etc.) derived from mammals including humans, and tissue sections. Therefore, the functional group may be a group capable of forming a covalent bond with a functional group in a protein, a sugar, or a nucleotide, for example, a hydroxyl group, an amino group, a hydroxyl group, a mercapto group, and the like.
  • each can be a carboxyl group, an aldehyde group, an amino group, a mercapto group, etc., and particularly, an aldehyde group that can form a covalent bond with an amino group by a mild reductive amination reaction is preferable.
  • an aldehyde group is used in a concept that also encompasses its protected form, the acetal or ketal form.
  • the polymer has a functional group with an lone pair at the other end that can be stably bonded to the surface of a solid substance composed of a metal or a semiconductor, or one or more functional groups with a lone pair at the side chain.
  • a lower alkyl group means a straight-chain or branched alkyl group having 1 to 6 carbon atoms, specifically, methyl, ethyl, propyl, and iso-alkyl. Examples thereof include propyl, petyl, sec.-butyl, tert.-butyl, pentyl, and hexyl. Therefore, examples of the sulfo lower alkyl include methylthio (one S CH 3 ) and ethyl thio (one SC 2 H 5 ).
  • a divalent group for example, alkylene
  • another polymer segment for example, polylactide, polypropylene oxide, etc.
  • the functional group is covalently linked as described above.
  • k is an integer from 1 to 8
  • cystinyl Cys—
  • methionyl Metal-
  • arginyl Arg—
  • the polymer segment having these residues in the side chain refers to, for example, a polymer segment having an amino acid residue as a repeating unit or a polyamine segment as described above. If necessary, these segments are covalently bonded to the PEG segment via a linking group (eg, one COO—, one CONH—, one NH—, etc.). Or another polyamine segment is also covalently linked to the PEG segment.
  • a linking group eg, one COO—, one CONH—, one NH—, etc.
  • another polyamine segment is also covalently linked to the PEG segment.
  • polymer in the context of the present invention is used to encompass not only the polymer commonly recognized by those skilled in the art but also the concept of an oligomer having several repeating units. are doing. Most of the above-mentioned polymers are known, and even if they are novel, they can be produced in the same manner as known polymers.
  • Representative examples of the polymer having a mercapto group at the terminal include those represented by the following formula (I).
  • Li, L 2 and L 3 independently represent a valence bond or Represents a linker, provided that when m is 0, 1 ⁇ and L 2 can be joined together to form a valence bond or one linker, and B is represented by the formula
  • R 1 R 2 CH 3 wherein R 1 and R 2 are independently a hydrogen atom, a lower alkyl group, and p is an integer from 2 to 5,
  • X represents a hydrogen atom, a hydroxyl group, a carboxyl group, an amino group or an aldehyde group, or a residue of one of the members capable of forming a specific binding pair, provided that X is a hydrogen atom Is L 3 is a valence bond, m is an integer from 0 to 100,000, and n is an integer from 10 to 20,000.
  • the specific binding pair has the meaning described above.
  • R 2 ′ / R 3 / wherein R 1 ′, R 2 / and R 3 ′ independently represent a lower alkyl group, L 3 ′ represents d-6 alkylene or a valence bond, and X ′ represents Represents a hydrogen atom, a hydroxyl group, a carboxyl group, an amino group, or an aldehyde group, or a one-member residue of a member capable of forming a specific binding pair, provided that when X ′ is a hydrogen atom, L 3 'is a valence bond and ⁇ m' is: Is an integer from 1 to 10,000, n 'is an integer from 10 to 20,000, and P' is an integer from 2 to 6.
  • the specific binding pair has the same meaning as defined for formula (I).
  • These polymers are conveniently prepared according to the methods described, for example, in Y. Nagasaki et al., Macromol. Rapid Commun. 1997, 18, 827 or K. Kataoka et al., Macromolecules 1999, 32. 6892-6894. Can be manufactured.
  • R 1 ′, R 2 ′ and R 3 ′ are each a methyl group
  • L 3 ′ —X ′ is —CH 2 CH 2 CHO
  • ⁇ ′ Wherein the polymer is 2.
  • the formation of the complex of the present invention is carried out at a temperature at which the solvent can maintain a liquid state in a solution capable of dissolving the polymer, for example, water, tetrahydrofuran, lower alkanol, or aromatic hydrocarbon.
  • a solution capable of dissolving the polymer for example, water, tetrahydrofuran, lower alkanol, or aromatic hydrocarbon.
  • the polymer and the polymer (functional groups capable of covalently bonding to one of the members forming the specific binding pair may be protected in some cases) may be contacted for a period of time sufficient to form the complex.
  • a polymer may be allowed to coexist when preparing the fine particles from a diluent such as a salt that can be a raw material of the fine particles.
  • 10 or more times, preferably 10 3 to 10 5 times, per atom or molar equivalent of the metal or semiconductor, and preferably 10 3 to 10 5 times of the polymer may be brought into contact with the metal or semiconductor.
  • the bonding ratio of the polymer is not limited, for example, when gold particles are used, it is preferable that the overnight potential of the entire composite is neutral particles within 1 mV of soil.
  • the conjugate thus prepared is protected, for example, when a functional group capable of covalently binding to one of the members forming a specific binding pair (for example, an aldehyde group: —CHO) is protected (for example, an acetal residue).
  • a functional group capable of covalently binding to one of the members forming a specific binding pair for example, an aldehyde group: —CHO
  • the group: _CH (OC 2 H 5 ) 2 ) may further have a protective group (acetal in this example) thereafter, if necessary, and may be covalently bonded to the one member.
  • the preparation of the present invention comprises the above polymer, if necessary, a buffer, a solvent, and, if the polymer has the above functional group, the above covalent bond, if necessary. It can be included together with a condensing agent that can be formed.
  • the preparation or composition that can be provided if the polymer has a functional group, as described above, for example, covalently binds an oligonucleotide, a probe, an antigen or an antibody as a probe to the complex.
  • This can provide a detection means that can be incorporated and used in a biological assay system.
  • a detection system capable of multiplex detection of complementary nucleic acids or fragments thereof can be constructed.
  • Fig. 1 shows a conceptual diagram of such a mode of use.
  • one of the members eg, a first antibody or complementary-strand DNA
  • an analyte eg, an antigen or DNA
  • a second antibody or a second complementary strand a portion not overlapping with the first complementary strand
  • the semiconductor-polymer When modified with a fluorescent dye, the semiconductor-polymer (first antibody or first complementary-strand DNA) one analyte (antigen or DNA)-A conjugate consisting of a second antibody or a second complementary chain is formed, and by selecting a combination of a semiconductor and a fluorescent dye, energy transfer from the semiconductor to the fluorescent dye occurs, and the fluorescent dye is Wavelength that emits light It is possible to emitting light to Konjiyugeto at different wavelengths.
  • the complex according to the invention can also be used in such a detection system. Therefore, according to the present invention, in a method for detecting an analyte (for example, an antigen or DNA) in a biological sample, the polymer in the complex containing the semiconductor particles of the present invention has a specific binding pair.
  • the first antibody or at least a part thereof has a first complementary strand having complementarity covalently bound to one end of a polymer as one of the members capable of forming a polymer, A complex comprising the union and the semiconductor, and a second complementary strand having complementation to a portion different from the second antibody or the first complementary strand to the analyte, the second complementary strand being modified with a fluorescent dye.
  • Detection characterized by combining with a biological sample and using the presence of luminescence at different wavelengths of the fluorescent dye generated by fluorescence resonance energy transfer from the semiconductor to the fluorescent dye as an indicator of the presence of the analyte Method provided it can.
  • fluorescent dyes that can be used in such combinations include, but are not limited to, Texas Red, FITC,
  • Alexa Fluor 532 (Molecular Probe), BOD I PY493 / 503 (Molecular Probe), BOD I PY FL (Molecular Probe), BOD I PY TR (Molecular Probe), Eosin (Molecular Probe), Fluorescein (Molecular Probe), Oregon Green 488 (Molecular Probe), Rhodamine Green dye (Molecular Probe), Rhodamine Red dye (Molecular Probe) and the like.
  • Example 1 Method for producing semiconductor particle composite:
  • the polymer used ( ⁇ -acetal-PEG-PAMA represented by the following formula):
  • the gold colloid-dolinate buffer aqueous solution (PH 7.5) was subjected to capillary electrophoresis under the same conditions as in Example 1. The results are shown in Figure 3.
  • Acetal-PEG-SH was produced as follows. In a receiver purged with argon, 20 m 1 of distilled tetrahydrofuran (THF) and 0.2 mmo 1 (0.032 ml) of initiator 3,3-dietkin-11-propanol were added, and an equivalent amount of potassium naphthalene was added. Was added and the mixture was stirred for 15 minutes to perform metallization.
  • THF distilled tetrahydrofuran
  • 0.2 mmo 1 0.032 ml
  • N-succinimidol_3- (2-pyridylthio) propionate (SPDP) 0.4 mmol (0.125 g) was dissolved in a small amount of distilled THF as a terminator. Was dropped with an equal pressure dropping funnel under ice-cooling. After stopping the reaction by stirring overnight, washing with saturated saline, extraction with chloroform, extraction with ether, freeze-drying of benzene, and The limer was recovered. The structure of the recovered polymer was confirmed by 1 H-NMR, and the amount of SPDP residues introduced into the terminal was determined by the UV absorption of 2-thiopyridone released by reacting with 2-mercaptoethanol. I also checked.
  • PEG-S S-Py 2 0x 10 - 2 mmo 1 (l O Omg) was dissolved in distilled water 4 m 1 and further five times mo 1 weight Jichiotorei torr 0. 1 mmo 1 a (15. 42 mg) The mixture was stirred at room temperature for 30 minutes. After the reaction, a polymer (hereinafter abbreviated as PEG 5000) was recovered through washing with saturated saline, extraction with chloroform, and reprecipitation with ether. The structure of the recovered polymer was confirmed by 1 H-NMR, and the terminal SH group was quantified by reaction with 2-pyridyl disulfide (21 PDS).
  • Texas Let's Door Avidin (Molecular Probe) 0.325 ⁇ L of CdS particles containing biotin-PEG-PAMA prepared in 0.3 mo 1 ZL solution and adding 0.025, 0.25 and 2.5 mmo 1 ZmL to the fluorescence spectroscopy The measurement was performed using an instrument (Hitachi Fluorometer F-2500) at an excitation wavelength of 400 nm. Fig. 4 shows the results.
  • a complex that can form a stable conjugate with an analyte in a biological sample and that can cause a change in light absorption wavelength or a change in fluorescence emission wavelength. Accordingly, the present invention may be utilized in the manufacturing industry of the detection sector or such detection reagent biological sample c

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Abstract

Complexes d'un métal ou d'un semi-conducteur comportant un polymère spécifique à sa couche de surface, utilisable dans un système pour détecter une substance test (un antigène, un ADN, etc.) dans un échantillon biologique.
PCT/JP2002/000121 2001-01-12 2002-01-11 Complexes d'un solide et de polymeres utilises dans un dosage biologique WO2002056020A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005010529A1 (fr) * 2003-07-28 2005-02-03 Tokyo University Of Science, Educational Foundation Surface de materiau de base rendue insensible a une adsorption non specifique
JP2006028354A (ja) * 2004-07-16 2006-02-02 Keio Gijuku 蛍光体及び蛍光体の製造方法
WO2007086189A1 (fr) * 2006-01-30 2007-08-02 Konica Minolta Medical & Graphic, Inc. Substance de marquage fluorescente de nanoparticules ou nanotiges
WO2008123291A1 (fr) * 2007-03-29 2008-10-16 Konica Minolta Medical & Graphic, Inc. Composé de marquage fluorescent

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11201971A (ja) * 1997-11-09 1999-07-30 Bayer Corp 流体試料中の分析対象物を検出する方法

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH11201971A (ja) * 1997-11-09 1999-07-30 Bayer Corp 流体試料中の分析対象物を検出する方法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BRUCHEZ M.: "Semiconductor nanocrystals as fluorescent biological labels", SCIENCE, vol. 281, 1998, pages 2013 - 2016, XP002125872 *
WILLARD D.: "CdSe-ZnS quantum dots as resonance energy transfer donors in a model protein-protein binding assay", NANO LETTERS, vol. 1, no. 9, February 2001 (2001-02-01), pages 469 - 474, XP002908558 *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005010529A1 (fr) * 2003-07-28 2005-02-03 Tokyo University Of Science, Educational Foundation Surface de materiau de base rendue insensible a une adsorption non specifique
US8841138B2 (en) 2003-07-28 2014-09-23 Jsr Corporation Surface of base material being inhibited in non-specific adsorption
US9862992B2 (en) 2003-07-28 2018-01-09 Jsr Corporation Surface of substrate onto which non-specific adsorption is restrained
JP2006028354A (ja) * 2004-07-16 2006-02-02 Keio Gijuku 蛍光体及び蛍光体の製造方法
WO2007086189A1 (fr) * 2006-01-30 2007-08-02 Konica Minolta Medical & Graphic, Inc. Substance de marquage fluorescente de nanoparticules ou nanotiges
JP5169223B2 (ja) * 2006-01-30 2013-03-27 コニカミノルタエムジー株式会社 ナノ粒子またはナノロッドからなる蛍光標識物質
WO2008123291A1 (fr) * 2007-03-29 2008-10-16 Konica Minolta Medical & Graphic, Inc. Composé de marquage fluorescent
JP5136548B2 (ja) * 2007-03-29 2013-02-06 コニカミノルタエムジー株式会社 蛍光体標識化合物

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