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WO2018056762A1 - Composition de réactif pour la mesure de l'albumine glyquée et procédé de mesure de l'albumine glyquée l'utilisant - Google Patents

Composition de réactif pour la mesure de l'albumine glyquée et procédé de mesure de l'albumine glyquée l'utilisant Download PDF

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WO2018056762A1
WO2018056762A1 PCT/KR2017/010499 KR2017010499W WO2018056762A1 WO 2018056762 A1 WO2018056762 A1 WO 2018056762A1 KR 2017010499 W KR2017010499 W KR 2017010499W WO 2018056762 A1 WO2018056762 A1 WO 2018056762A1
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albumin
dye
glycated albumin
measuring
boronic acid
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Korean (ko)
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이진우
전선아
박찬영
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Dxgen Corp
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Dxgen Corp
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Priority claimed from KR1020170122084A external-priority patent/KR102029798B1/ko
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Priority to EP17853476.4A priority Critical patent/EP3517963A4/fr
Priority to US16/335,412 priority patent/US20200018766A1/en
Publication of WO2018056762A1 publication Critical patent/WO2018056762A1/fr
Anticipated expiration legal-status Critical
Priority to US17/189,972 priority patent/US20210208152A1/en
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • 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
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • 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
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids

Definitions

  • the present invention relates to a reagent composition for measuring glycated albumin for diagnosing diabetes and a method for measuring glycated albumin using the same, and more specifically, a reagent for measuring glycated albumin including silica nanoparticle-boronic acid encapsulating dye. It relates to a composition and a method for measuring glycated albumin using the same.
  • Diabetes mellitus is a metabolic disease caused by an abnormality in insulin that plays a role in blood sugar control. Insulin production is insufficient or cannot be effectively used when insulin-producing cells are destroyed due to abnormal immune system. It is classified according to the cause of the onset, such as type 2 diabetes.
  • Diabetes is characterized by high blood glucose, which increases blood glucose levels, and failure to control blood glucose control can lead to complications such as diabetic retinopathy, kidney disease, and foot lesions. .
  • glucose oxidase which is used to measure glucose, is vulnerable to environmental effects such as pH or other interferences contained in blood, and may affect the activity of the enzyme as it generates hydrogen peroxide.
  • glycated hemoglobin (HbA1c) has been used as a biomarker for measuring glucose more accurately and more stably than glucose. Since glycated hemoglobin is stable until red blood cells disappear, it is used as an indicator of the average blood glucose level of 2 to 3 months. Therefore, it is used to investigate the progress of diagnosis and treatment of diabetes. However, glycated hemoglobin measurement is not suitable for some patients with abnormal blood glucose or patients with red blood cells, such as chronic renal failure.
  • Albumin is a protein that exists not only in the blood but also in major organs and body fluids. Depending on the concentration of glucose in the blood, glycated albumin is also formed. Albumin has a 10-fold higher binding rate of glucose than hemoglobin, and glycated albumin is more sensitive to glycemic changes than glycated hemoglobin, and albumin is 15-20 days shorter than red blood cell lifespan. The average value of blood glucose levels can be monitored. Therefore, it is useful as an important glycemic control indicator in diabetic patients with end-stage chronic renal failure patients, patients with iron deficiency anemia, and variants with hemoglobin.
  • U.S. Patent No. 77878989, U.S. Patent No. 6008006, U.S. Patent No. 8507223, China Patent Publication No. 104673878, and Chinese Patent Publication No. 104614459 relate to a conventional method for measuring glycated albumin and present in a sample.
  • Hydrogen peroxide (H 2 O 2 ) produced by the enzyme reaction of the glycosylated amino acid oxidase (EC 1.5.3) series that is specifically cleaved glycosylated protein to glycosylated amino acid or glycosylated peptide using protease
  • the glycosylated albumin enzymatic method was measured again using a peroxidase enzyme.
  • the glycated albumin enzyme method can test not only high selectivity and accuracy for albumin, but also faster than the immunoassay (10-30 minutes), but the activity of the enzyme is because the activity of the enzyme protein has a great effect on the efficiency of the glycated albumin assay. Strict attention is required to maintenance and storage.
  • the glycated albumin enzyme method must be preceded by a step for removing glycated amino acids and glycated peptides already present in the sample. Since glycated amino acid oxidase (EC 1.5.3) cannot use intact glycated proteins as a substrate, it is necessary to use proteolytic enzymes. It is a complex measurement consisting of a multi-step, such as the step of using a glycosylated protein to degrade glycated amino acids or peptides must be preceded. Therefore, there is a need to develop a simpler and faster method for measuring glycated albumin.
  • U.S. Pat.No.5223392, U.S. Pat.No.5908925, European Patent No.0657470, European Patent No.0257421 and Chinese Patent No.103554256 are albumin antibodies and peroxidase enzymes for detecting glycated albumin.
  • An enzyme immunoassay (ELISA) using a conjugated glycated albumin antibody was disclosed, and Ikeda et al.
  • an enzyme-boronic acid immunoassay (ELIBA) using an albumin antibody and peroxidase enzyme conjugated boronic acid. Ikeda et al, Clin Chem. 44 (2): 256-263, 1998.
  • US Patent No. 9128085, US Patent Publication No. 2006-0270060, US Patent Publication No. 2008-0227210, US Patent Publication No. 2010-0167306, US Patent No. 5470759 as Point of Care or Rapid Kit And US Patent No. 7659107 discloses a method using disposable strips and cassettes using antibody-based lateral flow immunochromatography to measure albumin and glycated albumin in samples of blood, saliva, and the like.
  • Patent No. 2014-0170766 discloses a method for preparing a rapid kit using lateral flow immunochromatography using albumin aptamer and glycated albumin aptamer derived from a nucleic acid, which acts similar to an antibody. Patent No.
  • 2014-0335630 discloses a measurement method using glycated albumin aptamer and surface plasmon resonance (SPR).
  • SPR surface plasmon resonance
  • U.S. Patent No. 5631364, U.S. Patent No. 7374943, and International Patent No. 2014-033258 disclose that a dye-bonded boronic acid derivative is reacted with glycated hemoglobin in blood, loaded and washed in a cartridge composed of porous filter paper, A method of determining the ratio of two substances by measuring the reflectance (% Reflectance) of total hemoglobin and dye-bound glycated hemoglobin was disclosed.
  • U.S. Patent No.5589393 discloses a method of fixing a boronic acid derivative to agarose beads to improve safety, and measuring glycated hemoglobin by reacting it with glycated hemoglobin in blood using a disposable cartridge.
  • 8557590 and Korean Patent No. 1128037 disclose a method of measuring glycated hemoglobin by directly loading a reacted sample into a cartridge composed of porous filter paper.
  • silica nanoencapsulates (a) a dye that specifically binds to whole albumin and (b) a dye that is complementary to the dye that specifically binds to whole albumin.
  • boronic acid conjugated with the particles it was confirmed that the amount of albumin and glycated albumin can be measured quickly and accurately by a simple and stable method using an optical device, and the present invention was completed.
  • An object of the present invention is to provide a reagent composition for measuring glycated albumin and a method for measuring glycated albumin using the same, which can easily and accurately diagnose the presence or absence of diabetes.
  • the present invention provides a saccharification comprising (a) a dye that specifically binds to whole albumin and (b) “silica nanoparticle-boronic acid encapsulating dye” that specifically binds to glycated albumin.
  • a saccharification comprising (a) a dye that specifically binds to whole albumin and (b) “silica nanoparticle-boronic acid encapsulating dye” that specifically binds to glycated albumin.
  • a reagent composition for albumin measurement is provided.
  • the present invention also comprises the steps of (a) adding a blood or plasma solution to a reagent comprising a "silica encapsulated silica nanoparticles-boronic acid encapsulating specifically with glycated albumin; (b) administering the reactants to an absorption pad of the cartridge followed by washing with a wash solution; (c) measuring the amount of glycated albumin by measuring the optical reflectivity of the absorbent pad with an optical instrument; (d) adding blood or plasma solution to a reagent containing a dye that specifically binds to whole albumin and reacting the same; (e) administering the reactants to an absorption pad of the cartridge followed by washing with a wash solution; (f) measuring the optical reflectivity of the absorbent pad with optics to determine the total amount of albumin; (g) providing a method for measuring glycated albumin, comprising calculating a ratio of glycated albumin based on the measured amounts of glycated albumin and total albumin.
  • the dye specifically binding to the whole albumin is characterized in that bromocresol green (Bromocresol green) or bromocresol purple (Bromocresol purple).
  • the dye encapsulated in the silica nanoparticles is a yellow-based or red-based dye
  • the yellow-based dye has an absorption wavelength of 400 ⁇ 430nm
  • the red dye has an absorption wavelength of 500 ⁇ 530nm do.
  • the yellow dye is tarrazine (Tartrazine)
  • the red dye is characterized in that the red 80.
  • sica nanoparticles encapsulated with dye is prepared by adding dye and silica to water and surfactant mixture or water and organic solvent mixture and stirring, followed by addition of basic catalyst. It is characterized by combining as.
  • the diameter of the "silica nanoparticles encapsulated dye” is characterized in that 10 ⁇ 500nm.
  • the "silica encapsulated silica nanoparticles-boronic acid” is a 4-carboxyphenyl boronic acid (CPBA) after amination of the "silica nanoparticles encapsulated dye” Or conjugated with, or “carboxylated silica nanoparticles” and then conjugated with 3-aminophenyl boronic acid (APBA).
  • CPBA 4-carboxyphenyl boronic acid
  • APBA 3-aminophenyl boronic acid
  • the optical device is characterized by measuring the optical reflectance by irradiating a light source simultaneously with the wavelength of the dye specifically binding to the whole albumin and the specific wavelength of "silica nanoparticle-boronic acid encapsulated dye" do.
  • the method for measuring glycated albumin is characterized by diagnosing diabetes according to the ratio of glycated albumin.
  • the reagent composition for measuring saccharified albumin according to the present invention includes “silica nanoparticle-boronic acid encapsulated with dye”, the absorption wavelength of the dye is not affected by pH, and has excellent stability even when stored for a month or more.
  • the amount of light absorbed by one particle is greater than that of one dye molecule, thereby accurately measuring the amount of glycated albumin in the blood having a low detection limit.
  • FIG. 1 is an explanatory diagram showing a manufacturing method of "silica nanoparticles encapsulated dye" according to an embodiment of the present invention.
  • Figure 2 is an explanatory view showing the coupling reaction of "silica nanoparticles-boronic acid encapsulated dye” and glycated albumin according to an embodiment of the present invention.
  • FIG. 3 is a flowchart illustrating a method for measuring glycated albumin and total albumin using the reagent composition and optical apparatus for measuring glycated albumin of the present invention.
  • Figure 4 is a graph showing the absorbance according to the absorption wavelength of the "dye encapsulated silica nanoparticles-boronic acid" prepared according to an embodiment of the present invention and the dye used.
  • Figure 5 is analyzed using a shape image and (B) dynamic light scattering method measured using (A) scanning electron microscope of "dyed encapsulated silica nanoparticles-boronic acid" prepared according to an embodiment of the present invention This is a graph about one size.
  • Figure 6 is a graph showing the reflectance value of "silica encapsulated silica nanoparticles-boronic acid" appear according to the concentration of glycated albumin of the plasma samples known concentration.
  • silica nanoparticles encapsulated with boronic acid conjugated to The instrument was intended to confirm that the amount of albumin and glycated albumin can be accurately measured in a simple and stable manner.
  • die encapsulated silica nanoparticles-boronic acid that specifically binds to glycated albumin was prepared, and a dye that specifically binds to whole albumin was added to prepare a reagent composition for measuring glycated albumin.
  • blood or plasma samples containing albumin and glycated albumin were added to each of the prepared reagent compositions, reacted with an absorbent pad, and then washed with unreacted dyes and impurities combined with impurities.
  • Optical reflectance of the absorbent pad was also measured to determine the amount of albumin and glycated albumin. As a result, it was confirmed that diabetes can be diagnosed simply and quickly through the ratio of glycated albumin.
  • the yellow tartarazine dye is encapsulated in the silica nanoparticles, and the hydroxyl group (-OH) on the surface is substituted with a primary amine group, and then 4-carboxyl is a glycated albumin binding material.
  • Bromocresol green a dye that binds 4-carboxylicphenyl-boronic acid (CPBA) to the surface to produce “silica nanoparticles-boronic acid encapsulating tartrazine” and binds specifically to albumin.
  • CPBA 4-carboxylicphenyl-boronic acid
  • the plasma samples filtered through the silica nanoparticles-boronic acid and bromocresol green encapsulating tarrazine are reacted, administered to an absorbent pad, washed, and then the absorbent pad is optically applied.
  • Glycosylation by irradiation with red (430 nm) and blue (630 nm) light sources to measure the optical reflectivity of glycated albumin labeled with “silica nanoparticles-boronic acid encapsulating tartrazine” and total albumin labeled with bromocresolgreen, respectively. It was confirmed that the ratio of albumin can be measured simply and quickly.
  • the present invention relates to a reagent composition for measuring glycated albumin comprising a dye that specifically binds albumin and “silica nanoparticle-boronic acid encapsulated with dyes” that specifically binds to albumin. .
  • the dye that specifically binds to the total albumin can be used without particular limitation as long as it is a dye that specifically reacts with albumin and glycated albumin, bromo having a 620 nm absorption wavelength band in blue at physiological neutral pH. Examples thereof include bromocresol green and violet, and bromocresol purple having a 580 nm absorption wavelength band.
  • the dye encapsulated in the silica nanoparticles is characterized by using a yellow-based or red-based dye having a complementary color relationship with the dye specifically binding to the whole albumin.
  • the yellow dye has an absorption wavelength of about 400 ⁇ 430nm
  • the red dye has an absorption wavelength of about 500 ⁇ 530nm
  • yellow tar dye (Tartrazine, 425nm)
  • red dye red 80 (red 80, 528 nm) and the like, but are not limited thereto.
  • sica nanoparticles encapsulating dye may be prepared by adding a dye and silica to a water and a surfactant mixture or a water and an organic solvent mixture, followed by stirring, and then adding a basic catalyst.
  • the surfactant is not particularly limited, but in the present invention, tritone x-100 or n-hexane may be used, and the silica may be exemplified by tetraethyl orthosilicate or tetramethyl orthosilicate. can do.
  • the basic catalyst is to promote the silica precursor encapsulates the dye, it may promote the hydrolysis of the silica precursor with water.
  • the silica precursor in the ionized state thus reacts with each other to release water or alcohol (ROH) and is connected to each other to form a silica network.
  • ROH water or alcohol
  • the basic catalyst may be exemplified by ammonium hydroxide, tetrapropylammonium chloride, tetrapropylammonium hydroxide, tetrabutylammonium bromide, tetrabutylammonium chloride or tetrabutylammonium hydroxide.
  • “Dye-encapsulated silica nanoparticles” can increase stability and sensitivity because dyes do not spill out, and are less biotoxic and can easily change surface functional groups.
  • the diameter of the "silica nanoparticles encapsulated dye” may be 10 ⁇ 500nm, it is preferably 30 ⁇ 100nm to maintain the unique properties of the dye. If it is less than 10 nm, the operation is difficult, and if it is more than 500 nm, the thickness becomes thick and the dye may appear cloudy.
  • Boronic acid derivatives to impart selectivity to glycated albumin to “silica nanoparticles encapsulated with dyes” include 4-carboxylicphenyl boronic acid (CPBA) and 3-aminophenyl boronic acid (3- Aminophenyl boronic acid (APBA) is preferred, where CPBA is used to amine “dye encapsulated silica nanoparticles”, and APBA is used to carboxylate “dye encapsulated silica nanoparticles” Conjugation may be via carbodiimide cross coupling. It is preferable to use CPBA because 4-carboxylicphenyl boronic acid (CPBA) has relatively higher thermal stability than 3-aminophenyl boronic acid (APBA). Do.
  • CPBA 4-carboxylicphenyl boronic acid
  • APBA 3-aminophenyl boronic acid
  • “Dye-encapsulated silica nanoparticles-boronic acid” can react with cis-diol of glycated albumin, and several dye molecules are encapsulated in silica nanoparticles, so that one dye Since the absorption of light is higher than that of molecules, the detection limit of glycated albumin can be improved.
  • the blood or plasma solution was reacted with “dye-encapsulated silica nanoparticles-boronic acid” capable of labeling glycated albumin, and then added dropwise to the cartridge to wash and record the reflectance value of glycated albumin. Then, react with a bromocresolgreen (BCG) solution that can stain the whole albumin, drop it into a cartridge and wash it to record the total albumin reflectivity value.
  • BCG bromocresolgreen
  • the percentage of glycated albumin is compared to the total albumin reflectivity compared to the reflectivity of glycated albumin. Can be calculated At this time, the total albumin can be measured first, and then the glycated albumin can be measured, but the percentage of glycated albumin can be calculated.
  • the present invention in another aspect, the method comprising the steps of: (a) adding a blood or plasma solution to a reagent containing "silica nanoparticles-boronic acid encapsulated dye" that specifically binds to glycated albumin; (b) administering the reactants to an absorption pad of the cartridge followed by washing with a wash solution; (c) measuring the amount of glycated albumin by measuring the optical reflectivity of the absorbent pad with an optical instrument; (d) adding blood or plasma solution to a reagent containing a dye that specifically binds to whole albumin and reacting the same; (e) administering the reactants to an absorption pad of the cartridge followed by washing with a wash solution; (f) measuring the optical reflectivity of the absorbent pad with optics to determine the total amount of albumin; (g) calculating a ratio of glycated albumin based on the measured amounts of glycated albumin and total albumin.
  • the optical device can be used without particular limitation as long as it can measure the optical reflectivity using optical properties, and the wavelength of the dye (blue or purple) that specifically binds to the whole albumin and the "dye (yellow) Or red light-encapsulated silica nanoparticles-boronic acid ”using a light source (eg, blue (630 nm) or red (430 nm)) capable of generating a specific wavelength at the same time, and reflecting the optical signal to the photodiode detector.
  • a light source eg, blue (630 nm) or red (430 nm)
  • the amount of albumin and glycated albumin can be measured by optical signal converter.
  • the ratio of glycated albumin can be calculated by the relative amount of glycated albumin according to the total albumin amount by the following equation.
  • Glycosylated albumin ratio (%) Glycosylated albumin / total albumin
  • the ratio of the glycated albumin is 16% or more can be diagnosed as diabetes.
  • Example 1 Preparation of silica nanoparticle-boronic acid (YD @ SNP-CPBA) encapsulated yellow dye
  • YD @ SNP For crosslinking of the carboxyl groups of YD @ SNP and CPBA, the hydroxyl group (-OH) on the surface of YD @ SNP was substituted with a primary amine group. That is, 100 mg of YD @ SNP was added to 100 ml of ethanol, dispersed for 30 minutes using an ultrasonic disperser, and then 1 ml of APTES (3-Aminopropyltriethoxysilane) was added to a stirrer and reacted at room temperature for 2 hours. After the reaction, 4 times ethanol wash and 3 times DI wash at 3800 rpm and 15 minutes using a centrifuge were placed in an oven at 60 ° C. and dried to prepare aminated YD @ SNP (YD @ SNP-NH 2 ). .
  • APTES 3-Aminopropyltriethoxysilane
  • EDC 1-ethyl-3 [3-dimethylaminopropyl] carbodiimide hydrochloride
  • CPBA 4-carboxylicphenyl-boronic acid
  • silica nanoparticle-boronic acid (YD @ SNP-CPBA) encapsulated a yellow dye by washing with ethanol 4 times and DI DI 3 times at 3800rpm, 15 minutes using a centrifuge at room temperature or lyophilization (YD @ SNP-CPBA). ) was prepared.
  • Example 2 YD @ SNP-CPBA and Bromocresol green Containing Saccharification Determination of glycated albumin using reagent composition for albumin measurement
  • Example 2 200 ⁇ l of the reagent composition (ZnCl 2 , NaCl, MgCl 2 , Triton X-100, NaN 3 , glycine, HEPES, pH 8.1) prepared in Example 1 was placed in a brown tube, and the reference reagent was prepared. (Asahi Kasei GA-L) was added to 5 ⁇ l of a plasma sample whose% value of glycated albumin was measured by an Olympus AU400 analyzer, followed by reaction for 2 minutes.
  • the reagent composition ZnCl 2 , NaCl, MgCl 2 , Triton X-100, NaN 3 , glycine, HEPES, pH 8.1
  • reaction solution 25 ⁇ l was placed on the cartridge absorption pad of optics (Episode ® 616, Dixsen) for 15 seconds, absorbed and 25 ⁇ l of washing solution (Morpholine, NaCl, Triton X-100, Glycerol, and NaN 3 mixture) was added. Wash for 15 seconds. Next, a cartridge for an optical device (episodes ® 616, Dix Xen) was measured optical reflectivity of the glycated albumin in the yellow.
  • optics Episode ® 616, Dixsen
  • reagent composition (Succinic acid, pH 5.5) containing bromocresol green was placed in a brown tube, 5 ⁇ l of the same plasma sample was added, and the reaction was carried out for 2 minutes. 25 ⁇ l of the reaction solution was placed on the cartridge absorption pad of optics (Episode ® 616, Dixsen) for 15 seconds, absorbed and 25 ⁇ l of washing solution (Morpholine, NaCl, Triton X-100, Glycerol, and NaN 3 mixture) was added. Wash for 15 seconds. Next, in the cartridge optics (episodes ® 616, Dix Xen) it was measured optical reflectivity of the total albumin in the blue.
  • The% reflectivity of glycated albumin was determined by comparing the optical reflectivity of glycated albumin measured in 2-1 and the optical reflectivity of total albumin measured in 2-2.
  • The% reflectance (% R) measured from each wavelength was converted into K / S value, which is a quantitative indicator of how much colored material is on the surface, and% reflectance is converted into K / S value.
  • the formula is:
  • the% reflectance value obtained by investigating the yellow light source representing the amount of glycated albumin and the% reflectance value obtained from the blue light source representing the total amount of albumin are respectively substituted by K / S values, and then the ratio is calculated by saccharification.
  • the amount of albumin could be measured.
  • the reagent composition for measuring glycated albumin according to the present invention includes a dye which can distinguish the total albumin and the glycated albumin, respectively, and thus, the glycated albumin can be easily obtained by using an optical analyzer through administration of a washing solution to a measurement cartridge without a separate separation process. Because it can be measured, it can be widely used to diagnose diabetes.

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Abstract

La présente invention concerne une composition de réactif pour la mesure de l'albumine glyquée permettant de diagnostiquer la présence ou l'absence d'un diabète et un procédé de mesure de l'albumine glyquée l'utilisant et, plus spécifiquement, une composition de réactif pour la mesure de l'albumine glyquée, la composition comprenant un complexe « acide boronique-nanoparticules de silice encapsulant un colorant », et un procédé de mesure de l'albumine glyquée l'utilisant. Le procédé de mesure de l'albumine glyquée selon la présente invention comprend les étapes suivantes : (a) introduction de sang ou d'une solution de plasma dans un réactif comprenant un complexe « acide boronique-nanoparticules de silice encapsulant un colorant » se liant spécifiquement à l'albumine glyquée, puis réaction ; (b) injection d'un produit de réaction dans un tampon absorbant d'une cartouche, puis lavage avec un liquide de lavage ; (c) mesure du facteur de réflexion optique du tampon absorbant à l'aide d'un instrument optique afin de déterminer la quantité d'albumine glyquée ; (d) introduction de sang ou d'une solution de plasma dans un réactif comprenant un colorant se liant spécifiquement à l'albumine totale, puis réaction ; (e) injection d'un produit de réaction dans un tampon absorbant d'une cartouche, puis lavage avec un liquide de lavage ; (f) mesure du facteur de réflexion optique du tampon absorbant à l'aide d'un instrument optique afin de déterminer la quantité d'albumine totale ; et (g) calcul de la proportion d'albumine glyquée sur la base des quantités mesurées d'albumine glyquée et d'albumine totale. Dans la composition de réactif pour la mesure de l'albumine glyquée selon la présente invention, le colorant a été encapsulé à l'intérieur de nanoparticules de silice, et par conséquent la longueur d'onde d'absorption inhérente du colorant n'est pas affectée par le pH, et la composition présente une excellente stabilité même lorsqu'elle est stockée pendant un mois ou plus.
PCT/KR2017/010499 2016-09-22 2017-09-22 Composition de réactif pour la mesure de l'albumine glyquée et procédé de mesure de l'albumine glyquée l'utilisant Ceased WO2018056762A1 (fr)

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EP17853476.4A EP3517963A4 (fr) 2016-09-22 2017-09-22 Composition de réactif pour la mesure de l'albumine glyquée et procédé de mesure de l'albumine glyquée l'utilisant
US16/335,412 US20200018766A1 (en) 2016-09-22 2017-09-22 Reagent composition for measuring glycated albumin and method for measuring glycated albumin using same
US17/189,972 US20210208152A1 (en) 2016-09-22 2021-03-02 Reagent composition for measuring glycated albumin and method for measuring glycated albumin using same

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KR10-2016-0121694 2016-09-22
KR20160121694 2016-09-22
KR1020170122084A KR102029798B1 (ko) 2016-09-22 2017-09-21 당화 알부민 측정용 시약 조성물 및 이를 이용한 당화 알부민의 측정방법
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US17/189,972 Division US20210208152A1 (en) 2016-09-22 2021-03-02 Reagent composition for measuring glycated albumin and method for measuring glycated albumin using same

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109991426A (zh) * 2019-04-03 2019-07-09 深圳市安帝宝科技有限公司 一种糖化血清白蛋白检测试剂盒
CN112430327A (zh) * 2020-11-25 2021-03-02 南开大学 一种网状磁性分子印迹共价有机框架材料及其制备方法与应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5919708A (en) * 1990-11-14 1999-07-06 Axis Biochemicals As Assay for glycated blood proteins
KR20080023280A (ko) * 2006-09-09 2008-03-13 삼성전자주식회사 폴리에틸렌글라이콜이 결합된 기능성 실리카 나노파티클 및그 제조방법
KR20100137851A (ko) * 2009-06-23 2010-12-31 주식회사 올메디쿠스 아조 보론산 염료를 이용한 당화단백질의 광학적 측정방법
WO2014033258A1 (fr) * 2012-08-31 2014-03-06 Axis-Shield Asa Méthode de dosage de l'hémoglobine glyquée
KR20160015495A (ko) * 2014-07-30 2016-02-15 케이맥(주) 다중 파장의 광을 이용한 혼합 시료의 분석 시스템

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5919708A (en) * 1990-11-14 1999-07-06 Axis Biochemicals As Assay for glycated blood proteins
KR20080023280A (ko) * 2006-09-09 2008-03-13 삼성전자주식회사 폴리에틸렌글라이콜이 결합된 기능성 실리카 나노파티클 및그 제조방법
KR20100137851A (ko) * 2009-06-23 2010-12-31 주식회사 올메디쿠스 아조 보론산 염료를 이용한 당화단백질의 광학적 측정방법
WO2014033258A1 (fr) * 2012-08-31 2014-03-06 Axis-Shield Asa Méthode de dosage de l'hémoglobine glyquée
KR20160015495A (ko) * 2014-07-30 2016-02-15 케이맥(주) 다중 파장의 광을 이용한 혼합 시료의 분석 시스템

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
See also references of EP3517963A4 *
WANG, JIASHENG ET AL.: "Silica-based Nanocomposites via Reverse Microemulsions: Classifications, Preparations, and Applications", NANOSCALE, vol. 6, no. 9, 2014, pages 4418 - 4437, XP055502107, DOI: doi:10.1039/c3nr06025j *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109991426A (zh) * 2019-04-03 2019-07-09 深圳市安帝宝科技有限公司 一种糖化血清白蛋白检测试剂盒
CN112430327A (zh) * 2020-11-25 2021-03-02 南开大学 一种网状磁性分子印迹共价有机框架材料及其制备方法与应用

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