WO2014122973A1 - Procédé de mesure d'une substance cible - Google Patents

Procédé de mesure d'une substance cible Download PDF

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Publication number
WO2014122973A1
WO2014122973A1 PCT/JP2014/050997 JP2014050997W WO2014122973A1 WO 2014122973 A1 WO2014122973 A1 WO 2014122973A1 JP 2014050997 W JP2014050997 W JP 2014050997W WO 2014122973 A1 WO2014122973 A1 WO 2014122973A1
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Prior art keywords
sample
target substance
surfactant
substance
vitamin
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PCT/JP2014/050997
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English (en)
Japanese (ja)
Inventor
内田　好昭
拓弥 佐久
和也 小見
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富士レビオ株式会社
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Priority to JP2014560702A priority Critical patent/JP6330666B2/ja
Publication of WO2014122973A1 publication Critical patent/WO2014122973A1/fr

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    • 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
    • G01N33/82—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving vitamins or their receptors

Definitions

  • the present invention relates to a method for measuring a target substance.
  • the target substance is measured for various tests such as clinical tests.
  • the target substance may be bound to the binding substance for the target substance in the sample to be measured.
  • Vitamin D hereinafter simply referred to as vitamin D
  • DBP vitamin D binding protein, also referred to as Gc globulin
  • a dissociation operation between vitamin D and DBP is required.
  • an organic solvent eg, ethanol, methanol, DMSO
  • a denaturant eg, acid, protein denaturant, surfactant, hydrolase
  • the present invention provides a method for measuring a target substance.
  • the present inventors have been able to accurately measure a target substance by treating a target substance-containing sample with a hydrophobic part composed of a hydrocarbon chain and a surfactant containing a hydrophilic part. As a result, the present invention has been completed.
  • the present invention is as follows.
  • Method for measuring target substance including: 1) treating the sample with a surfactant comprising a hydrophobic moiety composed of hydrocarbon chains and a hydrophilic moiety; and 2) detecting a target substance in the treated sample.
  • the hydrocarbon chain is a linear hydrocarbon group having 10 or more carbon atoms.
  • the hydrophilic moiety is a sulfonate group, a carboxylate group, or an ammonium group.
  • a kit for measuring a target substance comprising: 1) a hydrophobic part composed of a hydrocarbon chain and a surfactant containing a hydrophilic part; and 2) an affinity substance and / or a target substance preparation for the target substance.
  • the method of the present invention is useful for measuring a target substance. According to the present invention, a target substance can be measured quickly and easily.
  • the kit of the present invention is useful for simple implementation of the method of the present invention, for example.
  • the present invention provides a method for measuring a target substance in a sample.
  • the method of the present invention includes: 1) treating the sample with a surfactant comprising a hydrophobic moiety composed of hydrocarbon chains and a hydrophilic moiety; and 2) detecting a target substance in the treated sample.
  • Step 1 a sample is treated with a surfactant containing a hydrophobic part composed of hydrocarbon chains and a hydrophilic part.
  • a surfactant including a hydrophobic part composed of a hydrocarbon chain and a hydrophilic part may be abbreviated as “main surfactant” as necessary.
  • the sample used in the method of the present invention is a sample containing or suspected of containing a target substance.
  • the target substance include low molecular weight substances, proteins, sugars, nucleic acids (eg, DNA, RNA), and derivatives thereof.
  • the term “small substance” refers to a compound having a molecular weight of less than 1,500.
  • the low molecular weight substance is a natural substance or a synthetic substance.
  • the molecular weight of the low molecular weight substance may be less than 1,200, less than 1,000, less than 800, less than 700, less than 600, less than 500, less than 400, or less than 300.
  • the molecular weight of the low molecular weight substance may also be 50 or more, 100 or more, 150 or more, or 200 or more.
  • low molecular weight substances include ligands, hormones, lipids, fatty acids, vitamins, opioids, neurotransmitters (eg, catecholamines), nucleosides, nucleotides, oligonucleotides, monosaccharides, oligosaccharides, amino acids, and oligopeptides, or pharmaceuticals. , Toxicants, and metabolites.
  • hormones include steroid hormones, thyroid hormones, and peptide hormones.
  • the low molecular weight substance may be a vitamin.
  • vitamins include vitamins A, B1, B2, B6, B12, C, D, E, and K.
  • the vitamin is a fat-soluble vitamin (eg, vitamin A, D, E, K), more preferably vitamin D as shown below.
  • the low molecular weight substance may also be a metabolite of a vitamin.
  • vitamin metabolites include compounds having a hydroxyl group added to vitamins as described above, and conjugates (eg, glucuronic acid conjugates, sulfate conjugates, glutathione conjugates, acetyl conjugates, amino acid conjugates). Is mentioned.
  • the low molecular weight substance may further be a vitamin-like therapeutic drug or a metabolite thereof.
  • vitamin is intended to encompass vitamins and drugs similar to vitamins, and their metabolites, unless otherwise specified.
  • vitamin D is intended to encompass vitamin D2 and vitamin D3, and drugs similar to vitamin D2 and vitamin D3, and metabolites thereof.
  • the low molecular weight substance may be a steroid compound.
  • a steroid compound refers to a compound having a steroid skeleton.
  • Steroid compounds include steroid hormones and derivatives thereof that retain the steroid skeleton (eg, synthetic steroids such as anabolic steroids, anti-androgenic and anti-follicular hormone agents).
  • steroid hormones include male hormones, follicular hormones, luteinizing hormones, and corticoids (eg, glucocorticoids and mineralocorticoids), with follicular hormones being preferred.
  • Examples of follicular hormones include estrone, estradiol, and estriol.
  • the low molecular weight substance may also be a metabolite of a steroid compound.
  • the metabolite of the steroid compound include a compound in which a hydroxyl group is added to the steroid compound as described above, and a conjugate.
  • the conjugate for example, a glucuronic acid conjugate, a sulfate conjugate (eg, a hydroxyl group at either the 3-position or 17-position of estradiol, or a hydroxyl group at both the 3-position and 17-position was conjugated with a sulfate group).
  • Compound glutathione conjugates, acetyl conjugates, and amino acid conjugates.
  • the low molecular weight substance may further be a steroid compound-like therapeutic drug (eg, estramustine) or a metabolite thereof (eg, estromustine).
  • the low molecular weight substance may be an amino acid compound.
  • An amino acid compound refers to a compound having an amino group and a carboxyl group.
  • amino acid compounds include ⁇ -amino acids (eg, glycine, alanine, asparagine, cysteine, glutamine, isoleucine, leucine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, aspartic acid, glutamic acid, arginine, Histidine, lysine, ornithine, citrulline), ⁇ -amino acids (eg, ⁇ -alanine), ⁇ -amino acids (eg, ⁇ -aminobutyric acid), and derivatives thereof that retain amino and carboxyl groups.
  • ⁇ -amino acids eg, glycine, alanine, asparagine, cysteine, glutamine, isoleucine, le
  • the amino acid compound may be L-form or D-form.
  • the low molecular weight substance may also be a metabolite of an amino acid compound. Examples of the metabolite of the amino acid compound include a compound in which a hydroxyl group is added to the amino acid compound as described above, and a conjugate as described above.
  • the low molecular weight substance may further be an amino acid compound-like therapeutic drug or a metabolite thereof.
  • the amino acid compound may be a tyrosine derivative biosynthesized from tyrosine.
  • the tyrosine derivative include thyroid hormone (eg, triiodothyronine, thyroxine).
  • the tyrosine derivative may also be a metabolite of thyroid hormone.
  • thyroid hormone metabolites include compounds in which a hydroxyl group is added to thyroid hormone and conjugates as described above.
  • the low molecular weight substance may further be a thyroid hormone-like therapeutic drug or a metabolite thereof.
  • the origin of the sample is not particularly limited, and may be a biological sample derived from a living organism or an environmental sample.
  • organisms from which biological samples are derived include animals such as mammals (eg, humans, monkeys, mice, rats, rabbits, cows, pigs, horses, goats, sheep), birds (eg, chickens), insects, and the like. , Microorganisms, plants, fungi, and fish, preferably mammals, fungi, and fish, more preferably mammals, and even more preferably humans.
  • a biological sample can also be a blood-related sample (eg, whole blood, serum, plasma), saliva, urine, milk, tissue or cell extract, or a mixture thereof, which is the blood itself or a sample derived from blood.
  • a blood-related sample is preferred.
  • environmental samples include samples derived from soil, seawater, and fresh water.
  • the sample used in the method of the present invention is preferably a sample containing a complex of a target substance and a molecule capable of binding to the target substance.
  • the method of the present invention has an advantage that the target substance can be accurately measured even when a molecule (eg, protein) having a strong binding ability to the target substance exists in the sample.
  • a molecule eg, protein
  • the target substance in the sample can be accurately measured.
  • the target substance can be accurately measured according to the method of the present invention even when a molecule having an ability to bind to the target substance is present in the sample.
  • a sample containing a complex of a target substance eg, vitamin D such as vitamin D
  • a molecule capable of binding to the target substance include blood-related samples (eg, whole blood, serum, plasma). It is done.
  • the sample may be subjected to other treatments before being treated with the main surfactant.
  • treatment includes centrifugation, extraction, filtration, precipitation, heating, freezing, refrigeration, and stirring.
  • the volume of the sample to be treated with the main surfactant is not particularly limited as long as the target substance can be measured, but is, for example, 0.1 to 1000 ⁇ l, preferably 0.5 to 100 ⁇ l, more preferably 1 to 50 ⁇ l. .
  • the hydrophobic portion is composed of hydrocarbon chains.
  • the hydrocarbon chain is a linear or branched hydrocarbon group, and the number of carbon atoms in the hydrocarbon chain is usually 8 to 60.
  • the main surfactant need only have at least one such hydrocarbon chain.
  • the number of carbon atoms in the hydrocarbon chain is preferably 10 or more.
  • the number of carbon atoms in the hydrocarbon chain is also preferably 40 or less, more preferably 30 or less, and even more preferably 20 or less, from the viewpoint of easy synthesis or availability.
  • the number of carbon atoms in the hydrocarbon chain is particularly preferably 10 or 11.
  • linear hydrocarbon group examples include an alkyl group which is a linear saturated hydrocarbon group, and a linear unsaturated hydrocarbon group (eg, alkenyl group and alkynyl group).
  • alkyl group having 8 to 60 carbon atoms examples include octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl and icosyl.
  • Examples of the straight chain unsaturated hydrocarbon group having 8 to 60 carbon atoms include, for example, 1 to 4 (preferably 1 or 2) unsaturated bonds (double bonds or triple bonds) in the alkyl group described above. ) Part.
  • Examples of the branched hydrocarbon group include carbon atoms having 1 to 4 (preferably 1 or 2) carbon atoms having 1 to 10 carbon atoms on the above-described linear saturated or unsaturated hydrocarbon group. The thing substituted by the hydrogen group is mentioned.
  • hydrocarbon group having 1 to 10 carbon atoms examples include methyl, ethyl, propyl, iso-propyl, butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, cyclo Examples include propyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, phenyl, naphthalenyl.
  • the hydrocarbon chain constituting the hydrophobic portion is preferably a linear saturated hydrocarbon group, more preferably a linear saturated hydrocarbon group having 10 or 11 carbon atoms.
  • examples of the hydrophilic portion include an anionic hydrophilic portion and a cationic hydrophilic portion.
  • the surfactant including a hydrophobic part composed of a hydrocarbon chain and a hydrophilic part may have one or more hydrophilic parts, and includes an anionic hydrophilic part and a cationic hydrophilic part. You may have both.
  • examples of the anionic hydrophilic moiety include a sulfonate group (—SO 3 ⁇ ), a carboxylate group (—COO ⁇ ), and a phosphonate group (—POO 2 ⁇ ).
  • examples of the cationic hydrophilic portion include an ammonium group (N + ) and a phosphonium group (P + ).
  • the hydrophilic moiety is preferably a sulfonate group (—SO 3 ⁇ ), a carboxylate group (—COO ⁇ ), an ammonium group (N + ), or a phosphonium group (P + ), more preferably a sulfonate group ( —SO 3 ⁇ ), a carboxylate group (—COO ⁇ ), or an ammonium group (N + ), and even more preferably a sulfonate group (—SO 3 ⁇ ) or a carboxylate group (—COO ⁇ ). Particularly preferred is a sulfonate group (—SO 3 ⁇ ).
  • the main surfactant may contain other parts (for example, between the hydrophobic part and the hydrophilic part) in addition to the hydrophobic part and the hydrophilic part composed of the hydrocarbon chain.
  • Such other moieties include, for example, cyclic groups (eg, aryl groups such as cycloalkyl groups and phenyl groups, heterocyclic groups), and acyclic groups (eg, amino groups, oxy groups, oxo groups, carbonyl groups). , Carbonylamino group, carbonyloxy group).
  • the main surfactant may or may not be in the form of a salt.
  • the term “salt” is any salt, including, for example, inorganic salts, organic salts, and inner salts.
  • the inorganic salt include metal salts, halide salts, acid addition salts, and ammonium salts.
  • the metal salt include a salt of an alkali metal (eg, lithium, sodium, potassium) and a salt of an alkaline earth metal (eg, magnesium, calcium).
  • the halogen in the halide salt include fluorine, bromine, chlorine, and iodine.
  • Examples of acid addition salts that are inorganic salts include salts with inorganic acids such as hydrochloric acid, nitric acid, and sulfuric acid.
  • Examples of the organic salt include salts with organic bases such as trimethylamine, triethylamine and pyridine, and salts with organic acids such as oxalic acid.
  • examples of the main surfactant include hexyl sulfate, octyl sulfate, decyl sulfate, dodecyl sulfate, tetradodecyl sulfate, hexadecyl sulfate, dodecyl phosphonic acid, dodecyl benzene sulfonic acid, N-lauroyl sarcosine acid, and dodecacate. Noyl sarcosinic acid, as well as their salts.
  • Sample processing may be performed using one or more (eg, 2 or 3) primary surfactants.
  • the concentration of the main surfactant is not particularly limited as long as it is an effective concentration for the measurement of vitamin D, and can be adjusted as appropriate. For example, 0.001% (w / v) to 10% (w / v )% (W / v).
  • the concentration of such a surfactant when processing a sample with a main surfactant is a mixture of the reaction solution and the sample when the sample is processed by a method using a reaction solution as described later.
  • the concentration is not particularly limited as long as the concentration is such that its action is exerted, but may be, for example, 0.001% (w / v) to 10% (w / v).
  • the concentration of the main surfactant may be preferably 0.005% (w / v) or more, more preferably 0.01% (w / v) or more.
  • the concentration of the main surfactant is also preferably 8% (w / v) or less, more preferably 5% (w / v) or less, even more preferably 2% (w / v) or less, particularly preferably 1% It may be (w / v) or less.
  • the concentration of each surfactant is also as described above.
  • the concentration of the main surfactant in the case of processing the sample with the main surfactant is determined in the mixed liquid of the pretreatment liquid and the sample.
  • the concentration is not particularly limited as long as the effect is exhibited, but may be, for example, 0.001% (w / v) to 10% (w / v).
  • the concentration of the main surfactant is preferably 0.005% (w / v) or higher, more preferably 0.01% (w / v) or higher, even more preferably 0.1% (w / V) or more.
  • the concentration of the main surfactant is also preferably 8% (w / v) or less, more preferably 5% (w / v) or less, even more preferably 2% (w / v) or less, particularly preferably 1% It may be (w / v) or less.
  • concentration of each surfactant is also as described above. If it is intended to use the main surfactant at a lower concentration in the reaction solution, preferably dodecyl sulfate or a salt thereof may be used as the main surfactant. It has been confirmed that sodium dodecyl sulfate (SDS) exhibits a strong effect at low concentrations (see Tables 5 and 7).
  • Such a low concentration is a concentration of less than 0.1% (w / v), preferably less than 0.08% (w / v), more preferably 0.06% (w / v). ) Less than, and even more preferably less than 0.05% (w / v).
  • sample processing may be performed in combination with a reducing agent in addition to the main surfactant.
  • the method of the present invention may further comprise treating the sample with a reducing agent.
  • Treatment of the sample with the main surfactant and the reducing agent can be performed simultaneously or separately, but preferably is performed simultaneously.
  • Any reducing agent can be used as such a reducing agent, but a thiol group-containing reducing agent is preferred.
  • the thiol group-containing reducing agent include dithioerythritol, cysteine, 2-mercaptoethylamine, and dithiothreitol.
  • the concentration of the reducing agent is not particularly limited as long as it is an effective concentration for measuring the target substance, and can be adjusted as appropriate, but may be, for example, 0.01 to 1000 mM.
  • the concentration of the reducing agent when processing the sample with the reducing agent is such that when the sample is processed by a method using a reaction solution as described later, the effect is exerted in the mixed solution of the reaction solution and the sample.
  • the concentration is not particularly limited as long as the concentration is such that it may be, for example, 0.01 to 100 mM. In such a case, the concentration of the reducing agent may be preferably 0.05 mM or more, more preferably 0.1 mM or more.
  • the concentration of the reducing agent may also be preferably 100 mM or less, more preferably 50 mM or less, even more preferably 20 mM or less, and particularly preferably 10 mM or less.
  • concentration of each reducing agent is also as described above.
  • the concentration of the reducing agent when processing the sample with the reducing agent is effective when the sample is processed by a method using a pretreatment liquid as will be described later.
  • the concentration is not particularly limited as long as the concentration is such that it may be, for example, 0.1 to 100 mM. In such a case, the concentration of the reducing agent may be preferably 0.05 mM or more, more preferably 0.1 mM or more.
  • the concentration of the reducing agent may also be preferably 100 mM or less, more preferably 50 mM or less, and even more preferably 20 mM or less.
  • concentration of each reducing agent is also as described above.
  • the sample may be processed in combination with a main surfactant, or a combination of a main surfactant and a reducing agent, and another modifier different from the main surfactant.
  • the method of the present invention may further comprise treating the sample with another denaturing agent.
  • the treatment of the sample with the main surfactant, reducing agent and another denaturant can be done simultaneously or separately, but preferably is done simultaneously.
  • examples of such modifiers include surfactants (eg, anionic surfactants, cationic surfactants, amphoteric surfactants or nonionic surfactants) and chaotropic agents. Although 1 type may be sufficient as a modifier
  • denaturant multiple (for example, 2 or 3 types) may be sufficient as it.
  • Such a denaturant may be used at a concentration effective for the denaturing action, but may be used at a concentration effective for an action other than the denaturing action in anticipation of an action other than the denaturing action.
  • a surfactant having a steroid skeleton may be used as another denaturing agent different from the main surfactant.
  • the surfactant having a steroid skeleton is a compound having a steroid skeleton as an independent cyclic structure (that is, a steroid skeleton not condensed with another ring), or a salt thereof.
  • the three-dimensional structure at the 5-position may be ⁇ or ⁇ .
  • the concentration of the surfactant having a steroid skeleton is not particularly limited as long as it is an effective concentration for measurement of the target substance, and can be adjusted as appropriate. For example, 0.005% (w / v) to 10% (w / V), preferably 0.01% (w / v) to 1% (w / v).
  • the surfactant having a steroid skeleton may be a compound having a steroid skeleton as a hydrophobic moiety and a hydrophilic moiety or a salt thereof.
  • hydrophilic part of the surfactant having a steroid skeleton include an anionic part [eg, sulfonate (—SO 3 ⁇ ), carboxylate (—COO ⁇ ), and phosphonate (—POO 2 ⁇ )], cationic Moieties (eg, quaternary ammonium and quaternary phosphonium optionally substituted with 1 to 4 hydrocarbon groups), nonionic hydrophilic moieties (eg, multiple ethers) and groups having them (eg, this Hydrocarbon group having such a hydrophilic portion).
  • anionic part eg, sulfonate (—SO 3 ⁇ ), carboxylate (—COO ⁇ ), and phosphonate (—POO 2 ⁇ )
  • cationic Moieties eg
  • the surfactant having a steroid skeleton can be an anionic surfactant, a cationic surfactant, an amphoteric surfactant or a nonionic surfactant, depending on the type of the hydrophilic moiety.
  • the hydrocarbon group described above include methyl, ethyl, propyl, iso-propyl, butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl (lauryl).
  • the hydrocarbon group is preferably a hydrocarbon group having 1 to 10 carbon atoms, and more preferably an alkyl group having 1 to 6 carbon atoms.
  • the steroid skeleton may have 1 to 6 (preferably 1, 2, 3 or 4) substituents in addition to the hydrophilic portion.
  • a substituent is not particularly limited as long as it does not significantly impair the properties (eg, hydrophobicity) of the steroid skeleton, and examples thereof include a hydrocarbon group having 1 to 10 carbon atoms, a hydroxyl group, and 1 to 1 carbon atoms.
  • Hydroxyl group substituted with 10 hydrocarbon groups eg, alkyloxy group
  • hydrocarbon group having 1 to 10 carbon atoms-carbonyl-oxy group eg, alkyl-carbonyl-oxy group
  • oxo group formyl group
  • a hydrocarbon group having 1 to 10 carbon atoms-oxy-carbonyl group eg, alkyloxy-carbonyl group
  • halogen atom eg, fluorine atom, chlorine atom, bromine atom, iodine atom
  • the surfactant having a steroid skeleton is a bile acid or a derivative thereof or a salt thereof.
  • bile acids include deoxycholic acid, chenodeoxycholic acid, ursodeoxycholic acid, hyodeoxycholic acid, cholic acid, glycocholic acid, taurocholic acid, hyocholic acid, 5 ⁇ -cypurinol, lithocholic acid, taurodeoxycholic acid, and Taurocholic acid is mentioned.
  • bile acid derivatives include CHAPS, BIGCHAP, and deoxy-BIGCHAP.
  • the surfactant having a steroid skeleton may have a steroid skeleton having no hydroxyl group at the 7-position.
  • the surfactant having a steroid skeleton having no hydroxyl group at the 7-position for example, a surfactant having a steroid skeleton having a group other than the hydroxyl group (eg, the above-described substituent excluding the hydroxyl group) at the 7-position, And a surfactant having a steroid skeleton having no substituent at the 7-position (in other words, a carbon atom at the 7-position is bonded to a hydrogen atom).
  • the surfactant having a steroid skeleton having no hydroxyl group at the 7-position is a surfactant having a steroid skeleton having no substituent at the 7-position.
  • the surfactant having a steroid skeleton having no substituent at the 7-position include deoxycholic acid, taurodeoxycholic acid, lithocholic acid, 5 ⁇ -cyprinol, and salts thereof.
  • Sample processing may also be performed in the presence of other substances.
  • a substance include an affinity substance for a target substance, albumin (eg, bovine serum albumin, human serum albumin), gelatin, and skim milk.
  • the affinity substance for the target substance refers to a substance having an ability to bind to the target substance, and examples thereof include antibodies and aptamers for the target substance.
  • the antibody may be a polyclonal antibody or a monoclonal antibody.
  • the antibody may also be a fragment of an antibody (eg, Fab, F (ab ′) 2 ), a recombinant antibody (eg, scFv).
  • the antibody may further be an antibody-like molecule (eg, affibody, anticalin, DARPins, monobody) produced by molecular biological techniques such as phage display and / or by protein engineering techniques using existing protein motifs. May be.
  • Sample processing includes, for example, (a) mixing the sample with an aqueous solution (eg, buffer) containing the components as described above to prepare a mixture, and (b) incubating the mixture. .
  • aqueous solution eg, buffer
  • Tris buffer eg, Tris-HCl buffer, TE buffer, TAE buffer, TBE buffer, Tris buffered saline
  • phosphate buffer eg, phosphate buffered saline
  • Water e.g, carbonate-sodium bicarbonate buffer
  • GOOD buffer e.g, MES, ADA, PIPES, ACES, collamine hydrochloride, BES, TES, HEPES, acetamide glycine, tricine, glycinamide, bicine
  • the sample can be treated under neutral conditions or under acidic or alkaline conditions, but preferably under neutral conditions.
  • the pH value employed in the treatment of the sample is, for example, 4.0 to 9.5, preferably 5.0 to 9.0, preferably 5.5 to 8.5, more preferably 6.0 to 8. .0.
  • the pH value in the sample processing can be adjusted using a buffer solution, an acidic substance, and an alkaline substance.
  • the temperature of the sample treatment is not particularly limited as long as the components such as the main surfactant are suitable for exerting the action, but for example, 15 to 60 ° C.
  • the temperature is preferably 20 to 50 ° C, more preferably 20 to 45 ° C.
  • the time required for preparing the mixed solution in (a) is usually 30 seconds or less, preferably 20 seconds or less, more preferably 15 seconds or less, and even more preferably 10 seconds or less.
  • the incubation time in (b) is, for example, 60 minutes or less, preferably 30 minutes or less, more preferably 10 minutes or less.
  • the incubation time is still more preferably 5 minutes or less, particularly preferably 3 minutes or less, 2 minutes or less, 1 minute or less, 50 seconds or less, 40 seconds or less, 30 It may be 2 seconds or less, 20 seconds or less, 15 seconds or less, 10 seconds or less, or 5 seconds or less. Therefore, from the viewpoint of shortening the processing time for measurement, the sample processing time (eg, the total time of (a) and (b) above) is 3 minutes or less, 2 minutes or less, 1 minute or less, 50 seconds or less, It may be 40 seconds or less, 30 seconds or less, or 15 seconds or less.
  • Sample processing may also be performed by mixing alone.
  • the processing of the sample by “mixing only” means that the processing of the sample is performed by the above (a) and the above (b) is not performed (that is, incubation is unnecessary). From the viewpoint of quick and simple measurement, the sample may be processed only by mixing.
  • the above step 1) can be performed by treating the sample with a reaction solution containing a main surfactant and an affinity substance for the target substance.
  • the reaction solution contains a main surfactant and an affinity substance for the target substance.
  • the reaction solution may also contain a reducing agent.
  • the reaction solution further includes another modifying agent different from the main surfactant (eg, anionic surfactant, cationic surfactant, amphoteric surfactant or nonionic surfactant), a chaotropic agent, and a steroid skeleton.
  • another modifying agent different from the main surfactant eg, anionic surfactant, cationic surfactant, amphoteric surfactant or nonionic surfactant
  • a chaotropic agent e.g, a chaotropic agent
  • a steroid skeleton e.g, a steroid skeleton.
  • One or more (for example, 2 or 3 or more) of other components such as a surfactant having the above may be contained.
  • the concentration of the main surfactant in the reaction solution is not particularly limited as long as the concentration is such that the action is exerted in the mixed solution of the reaction solution and the sample. For example, 0.001 to 10% (w / v ).
  • the concentration of the main surfactant is preferably 0.005% (w / v) or more, more preferably 0.01% (w / v) or more.
  • the concentration of the main surfactant is also preferably 5% (w / v) or less, more preferably 3% (w / v) or less, even more preferably 1% (w / v) or less, particularly preferably 0.8. It may be 5% (w / v) or less.
  • the concentration of each major surfactant is also as described above.
  • the concentration of the reducing agent in the reaction solution is not particularly limited as long as the concentration is such that the action is exerted in the mixture of the reaction solution and the sample. For example, 0.001 to 1000 mM, preferably 0.01 to 100 mM. More preferably, it is 0.1 to 10 mM.
  • the concentration of the main surfactant in the reaction solution, and the concentration of the reducing agent, if included, is not particularly limited as long as the concentration is such that the action is exerted in the mixture of the reaction solution and the sample. In such a concentration that the above-mentioned concentration (main surfactant and / or reducing agent concentration when the sample is treated with the main surfactant and / or reducing agent) can be achieved in the mixture of the reaction solution and the sample. is there. Accordingly, the concentration of the main surfactant and / or the reducing agent in the reaction solution can be appropriately set so as to achieve the above-described concentration based on the volume of the sample and the reaction solution.
  • the reaction solution contains the above-described substances in an aqueous solution (eg, a buffer solution as described above).
  • the reaction solution may be a neutral solution, an acidic solution or an alkaline solution, but is preferably a neutral solution. Therefore, the pH value of the reaction solution is, for example, 4.0 to 9.5, preferably 5.0 to 9.0, preferably 5.5 to 8.5, and more preferably 6.0 to 8.0. .
  • the volume of the reaction solution can be appropriately determined according to the volume and type of the sample, the purpose of the assay (eg, qualitative or quantitative measurement), etc., but is 1 to 300 times, for example, preferably 1 to 150 times, more preferably 1 to 50 times.
  • the treatment of the sample with the reaction solution is appropriately performed in an appropriate manner to exert the action of components such as the main surfactant contained in the reaction solution.
  • the processing of the sample with the reaction liquid can be performed in the same manner as the processing of the sample as described above, (a1) mixing the sample with the reaction liquid to prepare a mixed liquid, and (b1) the mixed liquid. May be included.
  • the temperature and time conditions in (a1) and (b1) are the same as those described above in (a) and (b), respectively.
  • step 1) above comprises: i) treating the sample with a pretreatment liquid comprising a major surfactant; and ii) treating the sample treated with the pretreatment liquid with a diluent. It can be carried out.
  • the pretreatment liquid contains a main surfactant.
  • the pretreatment liquid may also contain a reducing agent.
  • the pretreatment liquid may further contain another modifier different from the main surfactant.
  • a surfactant having a steroid skeleton, and another surfactant different from the surfactant having a steroid skeleton eg, anionic surfactant, cationic surfactant, amphoteric surfactant
  • Agents or nonionic surfactants eg, anionic surfactant, cationic surfactant, amphoteric surfactant
  • Each of the main surfactant and another modifying agent contained in the pretreatment liquid may be one kind, but may be plural (eg, two or more kinds).
  • the pretreatment liquid is one or more (eg, 1 to 3) denaturing agents (eg, a surfactant having a steroid skeleton and another surfactant) that are the same type as the other denaturing agent contained in the diluent. May be included.
  • the pretreatment liquid may also contain other components as described above.
  • the concentration of the main surfactant in the pretreatment liquid, and when included, the concentration of the reducing agent is not particularly limited as long as the concentration is such that the action is exerted in the mixed liquid of the pretreatment liquid and the sample, For example, it is possible to achieve a concentration as described above (concentration of main surfactant and / or reducing agent when the sample is processed with the main surfactant and / or reducing agent) in the mixture of the pretreatment liquid and the sample.
  • concentration is high. Accordingly, the concentration of the main surfactant and / or the reducing agent in the pretreatment liquid can be appropriately set so as to achieve the above-described concentration based on the volume of the sample and the pretreatment liquid.
  • the pretreatment liquid contains the above-described substances in an aqueous solution (eg, a buffer as described above).
  • the pretreatment liquid may be a neutral solution, an acidic solution or an alkaline solution, but is preferably a neutral solution.
  • the pH value of the pretreatment liquid is, for example, 4.0 to 9.5, preferably 5.0 to 9.0, preferably 5.5 to 8.5, more preferably 6.0 to 8.0. is there.
  • the volume of the pretreatment liquid can be appropriately determined according to the volume and type of the sample, the purpose of the assay (eg, qualitative or quantitative measurement), etc., but is 0.5 to 100 times the sample volume, for example.
  • the ratio is preferably 1 to 10 times, more preferably 1 to 5 times.
  • the processing of the sample with the pretreatment liquid is appropriately performed in an appropriate manner to exert the action of the components contained in the pretreatment liquid.
  • the processing of the sample with the pretreatment liquid can be performed in the same manner as the processing of the sample as described above.
  • (A2-1) The sample is mixed with the pretreatment liquid to prepare the first mixed liquid, And (b2-1) incubating the first mixture.
  • the temperature and time conditions in (a2-1) and (b2-1) are the same as those described above in (a) and (b), respectively.
  • Processing of the sample with the pretreatment liquid may also be performed by mixing alone as described above.
  • the diluent may be, for example, the above-described components (eg, a surfactant having a steroid skeleton, an anionic surfactant, a cationic surfactant, an amphoteric surfactant, a nonionic surfactant, a chaotropic agent, and a reducing agent. ) May be included (for example, 2 or 3 or more types).
  • the diluent may also contain other components as described above (eg, affinity substance for vitamin D, albumin).
  • the concentration of the above-described components in the diluent is not particularly limited as long as the concentration is such that the action is exhibited in the pretreatment liquid, the diluent, and the mixed solution of the sample, and can be set as appropriate.
  • the diluent includes the above-described substances in an aqueous solution (eg, a buffer as described above).
  • the diluent may be a neutral solution, an acidic solution or an alkaline solution, but is preferably a neutral solution.
  • the pH value of the diluted solution is, for example, 4.0 to 9.5, preferably 5.0 to 9.0, preferably 5.5 to 8.5, and more preferably 6.0 to 8.0. .
  • the volume of the diluent can be appropriately determined depending on the volume and type of the sample and the pretreatment liquid, the purpose of the assay (eg, qualitative or quantitative measurement), etc., but is larger than the total volume of the sample and the pretreatment liquid.
  • the volume of the diluted solution is, for example, 1 to 20 times, preferably 1 to 10 times, more preferably 1 to 5 times the total volume of the sample and the pretreatment solution.
  • the processing of the sample with the diluent is appropriately performed in an appropriate manner to exert the action of the components contained in the diluent.
  • the sample processing with the diluent can be performed in the same manner as the sample processing with the pretreatment liquid.
  • the sample processing with the diluent includes (a2-2) preparing a second mixture by mixing the sample treated with the pretreatment liquid with the diluent, and (b2-2) second Incubating the mixture may include.
  • the temperature and time conditions in (a2-2) and (b2-2) are the same as those described above in (a) and (b), respectively.
  • the target substance is detected in the sample processed as described above.
  • the target substance is detected qualitatively or quantitatively.
  • it may include adding the affinity substance to the treated sample.
  • the target substance can be detected by any method, for example, using an affinity substance for the target substance.
  • the detection of the target substance may also be performed by an immunological technique.
  • immunological techniques include enzyme immunoassay (EIA) (eg, direct competition ELISA, indirect competition ELISA, sandwich ELISA), radioimmunoassay (RIA), fluorescence immunoassay (FIA), Examples thereof include a magnetic particle method, an immunochromatography method, a luminescence immunoassay method, a spin immunoassay method, and a latex agglutination method.
  • EIA enzyme immunoassay
  • RIA radioimmunoassay
  • FFA fluorescence immunoassay
  • a secondary antibody may be further used.
  • the secondary antibody may be an antibody against the primary antibody part of an antibody against the target substance (primary antibody), or an antibody against the complex of the target substance and the primary antibody.
  • An antibody such as a secondary antibody may be linked to a detection substance.
  • the detection substance examples include enzymes (eg, horseradish peroxidase, alkaline phosphatase), affinity substances (eg, streptavidin, biotin), fluorescent substances (eg, fluorescein, fluorescein isothiocyanate, rhodamine), luminescent substances (eg, , Luciferin, aequorin), radioactive substances (eg, 3 H, 14 C, 32 P, 35 S, 125 I).
  • the antibody such as a secondary antibody may be immobilized on a support.
  • the support include particles (eg, magnetic particles), membranes (eg, nitrocellulose membrane), glass, plastic, metal, plates (eg, multiwell plates), and devices.
  • the antibody may also be provided in a form impregnated in a medium such as filter paper.
  • the present invention also provides a kit for measuring a target substance.
  • the kit of the present invention comprises: 1) major surfactants; and 2) affinity substances for target substances and / or target substance preparations.
  • the kit of the present invention may also contain a reducing agent.
  • the kit of the present invention may further contain a modifying agent such as a surfactant having a steroid skeleton.
  • the affinity substance for the target substance is a primary antibody
  • the kit of the present invention may further contain a secondary antibody.
  • the kit of the present invention includes a reaction solution containing a main surfactant and an affinity substance for a target substance.
  • the reaction solution may further contain a reducing agent.
  • the kit of the invention comprises: 1) A pretreatment liquid containing a main surfactant (which may further contain a reducing agent); 2) Diluent; and 3) Affinity substance and / or target substance preparation for the target substance.
  • the reaction solution and the pretreatment solution and dilution solution may further contain the components and / or substances described above in the method of the present invention.
  • the target substance preparation is an aqueous solution or target substance powder containing a predetermined concentration (single or plural) of the target substance, and is useful as a control.
  • each component may be provided in a form accommodated in a different container (eg, tube, plate).
  • the kit of the present invention may be provided in the form of a device. Specifically, all of the components may be provided in a form housed in the device. Alternatively, some of the components may be provided in a form housed in the device, and the rest may be provided in a form that is not housed in the device (eg, a form housed in a different container). In this case, components that are not contained in the device may be used by being injected into the device during measurement of the target substance.
  • the structure of the device is, for example, 1) a first area for preparing a mixed solution by mixing a sample and a main surfactant, and contacting the prepared mixed solution with an affinity substance for a target substance.
  • a device comprising a second zone for detecting the target substance; 2) a device comprising a zone for detecting the target substance by mixing the sample, the main surfactant and an affinity substance for the target substance; and 3)
  • Examples of the device include a channel that allows mixing of a sample and the above-described components (eg, reaction solution, pretreatment solution and dilution solution), and an area for detecting a target substance.
  • 25OH vitamin D2 and 25OH vitamin D3 are used as immunological methods for measuring vitamin D as a target substance.
  • the primary antibody that recognizes (abbreviated as) was used.
  • the values measured in the following examples may correspond to the total amount of 25OH vitamin D2 and 25OH vitamin D3.
  • Example 1 Examination of 25OH Vitamin D Measurement Method Using Anionic Surfactant and Reducing Agent Human serum was treated with anionic surfactant (SDS) and reducing agent (DTT) and then humanized by immunological methods. Serum 25OH vitamin D was measured.
  • SDS anionic surfactant
  • DTT reducing agent
  • the method was performed as follows. 1) To 10 ⁇ l of the same serum collected from the same human, add 1%, 0.30%, 0.1% or 0% (w / v) SDS + 8 mM, 4 mM in PBS (pH 7.6) 40 ⁇ l of 2 mM or 0 mM DTT + 0.1% (w / v) deoxycholate sodium salt + 0.1% (w / v) CHAPS] was added to prepare a first mixed solution (50 ⁇ l) of serum and pretreatment solution. . The concentration of each component in the first mixture is as follows: 0.8%, 0.24%, 0.08% or 0% (w / v) SDS; 6.4 mM, 4 mM, 1.
  • Example 2 Investigation of effective reducing agents to denature vitamin D binding protein Human serum is treated with an anionic surfactant (SDS) and various reducing agents and then immunologically treated in human serum Of 25OH vitamin D was measured.
  • SDS anionic surfactant
  • the method was performed as follows. 1) To 10 ⁇ l of the same serum collected from the same human, pretreatment solution [PBS (pH 7.6) 0.3% (w / v) SDS + 5 mM each reducing agent + 0.1% (w / v) deoxycholic acid 40 ⁇ l of sodium salt + 0.1% (w / v) CHAPS] was added to prepare a first mixed solution (50 ⁇ l) of serum and pretreatment solution. The concentration of each component in the first mixture is as follows: 0.24% (w / v) SDS; 4 mM each reducing agent; 0.08% (w / v) deoxycholate sodium salt; and 0.08% (w / v) CHAPS. 2) Subsequent operations were performed in the same manner as in the above 2) to 12) of Example 1.
  • Example 3 Examination of treatment time with anionic surfactant (SDS) The treatment time with an anionic surfactant (SDS), which is required for highly sensitive measurement of 25OH vitamin D, was examined.
  • SDS anionic surfactant
  • the method was performed as follows. 1 Pre-treatment solution (0.3% (w / v) SDS + 5 mM 2-MEA + 0.1% (w / v) deoxycholate sodium salt + 0.1% in PBS was added to 10 ⁇ l of the same serum collected from the same human. w / v) 40 ⁇ l of CHAPS) was added, and a first mixed solution (50 ⁇ l) of serum and pretreatment solution was prepared by mixing by stirring. The concentration of each component in the first mixture is as follows: 0.24% (w / v) SDS; 4 mM 2-MEA; 0.08% (w / v) deoxycholate sodium salt; and 0.08% (w / v) CHAPS. The time required for mixing by stirring was about 3 seconds.
  • the luminescence count measured under the condition of mixing alone (incubation time: 0 min) was almost the same as the luminescence count measured under the condition of 10 min incubation time.
  • the incubation operation in the above step 2) is not necessarily required, and vitamin D is dissociated from DBP only by gently stirring the serum and the pretreatment liquid. Therefore, it was shown that the time required for measuring the target substance can be shortened by the method of the present invention.
  • Reference Example 1 Time required for pretreatment using an existing product
  • the time required for pretreatment of a serum sample is as follows according to its instruction manual.
  • Existing products include DiaSorin-RIA (DiaSorin), DiaSorin-Liaison (DiaSorin)
  • Example 4 Comparison between the method of the present invention and an existing method using an organic solvent (I) In order to verify the measurement accuracy of the target substance by the method of the present invention using an anionic surfactant (SDS), the measured value of the target substance by the method of the present invention and the measured value obtained by the existing method using an organic solvent And the correlation coefficient was compared.
  • SDS anionic surfactant
  • the concentration of each component in the mixture is as follows: about 0.03% (w / v) SDS; about 0.5 mM 2-MEA; about 0.04% (w / v) sodium deoxycholate About 0.04% (w / v) CHAPS; 2) The mixture was incubated at 37 ° C. for 10 minutes. 3) After incubation, the magnetic particles in the sample were collected on the magnetic plate, and the magnetic particles were washed three times. 4) Alkaline phosphatase labeled antibody (antibody against 25OH vitamin D-anti-25OH vitamin D antibody immune complex) suspended in MES buffer was added to the magnetic particles excluding the washing solution. 5) The solution obtained in the above 4) was incubated at 37 ° C. for 10 minutes.
  • Luminescence count was measured with a label reader (ARVO; Perkin Elmer).
  • Example 5 Comparison between the method of the present invention and an existing method (DiaSorin-RIA) (II) In order to further verify the measurement accuracy of the target substance by the method of the present invention using an anionic surfactant (SDS), the measured value of the target substance by the method of the present invention and the existing method (DiaSorin-RIA) were obtained. The correlation coefficient with the measured value was compared.
  • SDS anionic surfactant
  • DiaSorin-RIA Measurement by an existing method (DiaSorin-RIA) As an existing method, DiaSorin-RIA was used as described later. DiaSorin-RIA was performed using a commercially available kit (25-Hydroxyvitamin D 125I RIA Kit, manufactured by DiaSorin). The method was performed as follows. As a measurement sample, the same 13 human serum samples as in 5-1) above were used. I) Pretreatment operation a) A glass test tube is prepared. b) Dispense 500 ⁇ l of acetonitrile into each test tube. c) Add 50 ⁇ l of calibrator, control, or sample (such as serum). d) Stir the sample solution for 10 seconds. e) Centrifuge the sample solution at 1200 g for 10 minutes at room temperature. f) The supernatant is used as a sample.
  • a measurement sample the same 13 human serum samples as in 5-1) above were used. I) Pretreatment operation a) A glass test tube is prepared. b) Dispense 500 ⁇ l
  • Example 6 Measurement of vitamin D with SDS-like surfactant Next, the measurement of vitamin D with an SDS-like surfactant was examined.
  • the method was performed as follows. 1) 50 ng / ml fortified horse serum 3.75 ⁇ l, reaction solution [Tris-HCl buffer (pH 7.6), 0.03% (w / v) or 0.1% (w / v)] Agent + 0.038% (w / v) deoxycholate sodium salt + 0.038% (w / v) BSA + anti-25OH vitamin D antibody-binding particle solution] 146.25 ⁇ l was added, and a mixture of serum and reaction solution ( 150 ⁇ l) was prepared.
  • the concentration of each component in the mixture is as follows: about 0.03% (w / v) or about 0.1% (w / v) The following surfactant; about 0.04% (w / v) v) sodium deoxycholate; and about 0.04% (w / v) BSA. 2) Subsequent operations were performed in the same manner as in 2) to 9) of Example 4-1).
  • Example 7 Measurement of vitamin D by a surfactant having a hydrocarbon chain Next, measurement of vitamin D by a surfactant having a hydrocarbon chain was examined.
  • the method was as follows. 1) The reaction solution [Tris-HCl buffer (pH 7.6), 0.03% (w / v) or 0.1% (w / v)] was added to 3.75 ⁇ l of 100 ng / ml fortified horse serum. Agent, 0.038% (w / v) sodium deoxycholate, 0.038% (w / v) BSA, anti-25OH vitamin D antibody-binding particle solution] 146.25 ⁇ l is added, and the serum and reaction mixture are mixed A liquid (150 ⁇ l) was prepared.
  • the concentration of each component in the mixture is as follows: about 0.03% (w / v) or about 0.1% (w / v) The following surfactant; about 0.04% (w / v) v) sodium deoxycholate; and about 0.04% (w / v) BSA. 2) Subsequent operations were performed in the same manner as in 2) to 9) of Example 4-1).
  • the method and kit of the present invention are useful for measuring a target substance.

Abstract

La présente invention concerne un procédé de mesure d'une substance cible. Plus particulièrement, la présente invention concerne un procédé de mesure d'une substance cible qui consiste à : (1) traiter un échantillon avec un tensioactif comprenant une fraction hydrophile et une fraction hydrophobe constituée par une chaîne hydrocarbonée; et (2) détecter la substance cible dans l'échantillon traité. La présente invention concerne également un kit de mesure de substance cible qui comprend : (1) un tensioactif comprenant une fraction hydrophile et une fraction hydrophobe constituée par une chaîne hydrocarbonée; et (2) une substance qui a une affinité pour la substance cible, et/ou une préparation de la substance cible.
PCT/JP2014/050997 2013-02-06 2014-01-20 Procédé de mesure d'une substance cible WO2014122973A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018047792A1 (fr) * 2016-09-06 2018-03-15 富士レビオ株式会社 Méthode et réactif pour la mesure de thyroglobuline
WO2018051965A1 (fr) * 2016-09-13 2018-03-22 富士レビオ株式会社 Procédé de dosage de troponine cardiaque et réactif de dosage
CN108918848A (zh) * 2018-05-22 2018-11-30 德康润生物科技（北京）有限公司 维生素d释放剂及其制备方法与应用
WO2020241443A1 (fr) * 2019-05-24 2020-12-03 富士レビオ株式会社 Méthode et réactif pour la mesure de la thyroglobuline

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000241429A (ja) * 1998-07-31 2000-09-08 Mitsubishi Chemicals Corp 生理活性成分の測定法
JP2009236810A (ja) * 2008-03-28 2009-10-15 Itea Inc 食品検査方法及び食品抽出液処理用試薬
JP2010122205A (ja) * 2008-08-29 2010-06-03 Sysmex Corp 麻疹ウイルス検出方法、メンブレンアッセイ用試験具およびメンブレンアッセイ用試験キット
WO2012091569A1 (fr) * 2010-12-28 2012-07-05 Future Diagnostics B.V. Réactif de libération pour la vitamine d

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3487642B2 (ja) * 1994-06-16 2004-01-19 アボットジャパン株式会社 Ｈａｖ抗原及びその抗原と免疫学的に反応性の抗体の測定法
JP2008275534A (ja) * 2007-05-02 2008-11-13 Olympus Corp 標的高分子の精製方法及び検出方法
US7790401B2 (en) * 2007-08-06 2010-09-07 Siemens Healthcare Diagnostics Methods for detection of immunosuppressant drugs

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000241429A (ja) * 1998-07-31 2000-09-08 Mitsubishi Chemicals Corp 生理活性成分の測定法
JP2009236810A (ja) * 2008-03-28 2009-10-15 Itea Inc 食品検査方法及び食品抽出液処理用試薬
JP2010122205A (ja) * 2008-08-29 2010-06-03 Sysmex Corp 麻疹ウイルス検出方法、メンブレンアッセイ用試験具およびメンブレンアッセイ用試験キット
WO2012091569A1 (fr) * 2010-12-28 2012-07-05 Future Diagnostics B.V. Réactif de libération pour la vitamine d

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018047792A1 (fr) * 2016-09-06 2018-03-15 富士レビオ株式会社 Méthode et réactif pour la mesure de thyroglobuline
JPWO2018047792A1 (ja) * 2016-09-06 2019-06-24 富士レビオ株式会社 サイログロブリンの測定方法及び測定試薬
JP7007278B2 (ja) 2016-09-06 2022-02-10 富士レビオ株式会社 サイログロブリンの測定方法及び測定試薬
US11768209B2 (en) 2016-09-06 2023-09-26 Fujirebio Inc. Method and reagent for measuring thyroglobulin
WO2018051965A1 (fr) * 2016-09-13 2018-03-22 富士レビオ株式会社 Procédé de dosage de troponine cardiaque et réactif de dosage
JPWO2018051965A1 (ja) * 2016-09-13 2019-06-24 富士レビオ株式会社 心筋トロポニンの測定方法及び測定試薬
JP6999561B2 (ja) 2016-09-13 2022-02-10 富士レビオ株式会社 心筋トロポニンの測定方法及び測定試薬
US11802867B2 (en) 2016-09-13 2023-10-31 Fujirebio Inc. Cardiac troponin assay method and assay reagent
CN108918848A (zh) * 2018-05-22 2018-11-30 德康润生物科技（北京）有限公司 维生素d释放剂及其制备方法与应用
WO2020241443A1 (fr) * 2019-05-24 2020-12-03 富士レビオ株式会社 Méthode et réactif pour la mesure de la thyroglobuline
CN113631924A (zh) * 2019-05-24 2021-11-09 富士瑞必欧株式会社 甲状腺球蛋白的测定方法和测定试剂

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