WO2006054519A1 - Article standard pour détermination fractionnelle de composant de lipoprotéine - Google Patents

Article standard pour détermination fractionnelle de composant de lipoprotéine Download PDF

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Publication number
WO2006054519A1
WO2006054519A1 PCT/JP2005/020845 JP2005020845W WO2006054519A1 WO 2006054519 A1 WO2006054519 A1 WO 2006054519A1 JP 2005020845 W JP2005020845 W JP 2005020845W WO 2006054519 A1 WO2006054519 A1 WO 2006054519A1
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WO
WIPO (PCT)
Prior art keywords
acid
lipoprotein
measured
component
standard product
Prior art date
Application number
PCT/JP2005/020845
Other languages
English (en)
Japanese (ja)
Inventor
Isami Tsuboi
Michihiro Taguchi
Akira Miike
Original Assignee
Kyowa Medex Co., Ltd.
Bml, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kyowa Medex Co., Ltd., Bml, Inc. filed Critical Kyowa Medex Co., Ltd.
Priority to JP2006545011A priority Critical patent/JP4704355B2/ja
Publication of WO2006054519A1 publication Critical patent/WO2006054519A1/fr

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Classifications

    • 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
    • G01N33/92—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving lipids, e.g. cholesterol, lipoproteins, or their receptors
    • 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
    • G01N33/96—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving blood or serum control standard

Definitions

  • the present invention relates to a standard product used for differential quantification of a component to be measured in lipoprotein, a method for producing the same, a kit for fractional quantification of a component to be measured in a sample containing lipoprotein, and a lipoprotein.
  • the present invention relates to a method for fractional quantification of components to be measured in a sample containing protein.
  • lipid items For clinical diagnosis, measurement of lipid items is important for prevention of lifestyle-related diseases such as diabetes and improvement of lifestyle habits. Many methods for measuring lipid items in a specimen have been reported, but the so-called homogenous method, in which components in the lipoprotein are measured without separating the lipoprotein in the specimen, has become common. ing. For example, in a discrete type automatic analyzer, a large amount of sample is continuously quantified by a homogenous method using a surfactant such as dextran sulfate or a homogen method using a surfactant.
  • a surfactant such as dextran sulfate
  • surfactant such as dextran sulfate
  • the concentration of a component to be measured in a sample is calculated by measuring a standard product containing a measurement target component at a known concentration at the same time as the sample and analyzing the absorbance obtained from the standard product. This is done by using a calibration curve or correlation equation between the amount of information and the amount of information such as absorbance obtained from the specimen.
  • the components to be measured are individually contained at a constant concentration or content, so-called pure type standard products, as well as other components in the sample to be measured.
  • a product form containing a known concentration or a known content together with a component, a so-called serum type standard product, is known.
  • a standard product described in Patent Document 1 is known as a serum type standard product.
  • Patent Document 2 discloses lipoproteins for diagnostic purposes that have a water content in the range of 1 to 10% and can be reconstituted into a clear solution with a maximum turbidity of 70 LSU.
  • a lyophilized control or reference plasma or control or reference serum is disclosed.
  • a standard product of pure product type for example, when preparing a standard product by dissolving cholesterol or neutral fat, it is necessary to use a large amount of surfactant. As the force changes, pipetting with an automatic analyzer causes problems such as sucking up a larger amount of solution and not giving accurate measurements.
  • Patent Document 1 Japanese Patent Laid-Open No. 10-17597
  • Patent Document 2 JP-A-6-300758
  • An object of the present invention is to provide a standard product for fractional quantification of a component to be measured in a lipoprotein which is easy to handle and gives an accurate measurement value even after long-term storage, a method for producing the same, and a lipoprotein. It is an object of the present invention to provide a kit for fractional quantification of components to be measured in a contained sample and a method for fractional quantification of components to be measured in a sample containing lipoproteins. Means for solving the problem
  • the present invention is (1) a standard product obtained by freeze-drying serum or plasma to which a buffer having a sulfo group is added, which is used for differential quantification of components to be measured in lipoproteins.
  • a method for producing a lyophilized standard used for differential quantification of a component to be measured in lipoprotein characterized in that the content of the component to be measured is determined using a known amount of the component to be measured (7) Buffering power with sulfo group N- (2 Hydroxyethyl) N, One (2-sulfethenole) piperazine, Piperazine one N, N, One bis (2-ethanesnorephonic acid) , N— (2-acetamido) 2 aminoethanesulfonic acid, 3 morpholino-2-hydroxypropane sulfonic acid, N, N bis (2 hydroxyethyl) 2 aminoethanesulfonic acid, 3 morpholinopropane sulfonic acid, N, N bis (2 hydroxyethyl) 2 aminoethanesulfonic acid, 3 morpholinopropane sulfonic acid, N
  • Step [2] Calibration curve using the standard product according to any one of (1) to (5) above and the reagent for fractionation measurement And [3] calculating the concentration of the component in the sample from the measurement value obtained in the step [1] and the calibration curve created in the step [2].
  • the present invention relates to a method for fractional quantification of a component to be measured in a specimen containing a characteristic lipoprotein.
  • a standard product for fractional quantification of components to be measured in a lipoprotein that is easy to handle and gives an accurate measurement value even after long-term storage.
  • the reagent for fractional measurement of the component to be measured it is possible to accurately perform the fractional quantification of the component to be measured in the sample containing lipoprotein.
  • the specimen containing the component to be measured in the present invention is not particularly limited as long as it contains at least a plurality of various lipoproteins.
  • whole blood, plasma, serum, joint fluid, spinal fluid, saliva, amniotic fluid Urine, sweat, spleen, synovial fluid, etc. are preferred.
  • lipoprotein in the present invention examples include high density lipoprotein (hereinafter referred to as HDL), low density lipoprotein (hereinafter referred to as LDL), very low density lipoprotein (hereinafter referred to as VLDL), and chylomicron (hereinafter referred to as HDL). , CM)) and remnant lipoproteins.
  • HDL high density lipoprotein
  • LDL low density lipoprotein
  • VLDL very low density lipoprotein
  • HDL chylomicron
  • remnant lipoproteins remnant lipoproteins.
  • the components to be measured in the lipoprotein in the present invention include, for example, cholesterol in HDL (HDL cholesterol), neutral fat in HDL, cholesterol in LDL (LDL cholesterol), neutral fat in LDL, Examples include cholesterol in VLDL, neutral fat in VLDL, cholesterol in CM, neutral fat in CM, cholesterol in remnant lipoprotein, and neutral fat in remnant lipoprotein.
  • serum or plasma means the normal meaning of serum or plasma obtained by removing blood cell components and the like from blood.
  • blood blood derived from animals such as humans is used.
  • the buffer having a sulfo group in the present invention is preferably a buffer that gives a pH at which lipoproteins in serum or plasma do not precipitate.
  • pH 6-9 is preferable.
  • the buffer containing a sulfo group include N— (2-hydroxyethyl) N, one (2—snolechechinole) piperazine (HEPES), piperazine one N, N, one bis (2-ethanesulphonic acid).
  • POPSO N— (2acetamido) 2 aminoethanesulfonic acid
  • MOPSO morpholino 2-hydroxypropanesulfonic acid
  • BES N-bis (2-hydroxychetetyl) 2 aminoethanesulfonic acid
  • MOPS morpholinopropanesulfonic acid
  • TES Tris (hydroxymethyl) methyl] -2-aminoethanesulfonic acid
  • DIPSO N-bis (2hydroxyethyl) amino] — 2 Hydroxypropane sulfonic acid
  • TAPSO Piperazine 1 N, N, 1 bis (2-hydroxypropane sulfonate) Acid) (POPSO), 3- [4- (2 Hydroxyethyl) 1-piperazyl] -2 Hydroxypropane sulfonic acid
  • HEPP Piperazine 1 N, N, 1 bis (2-hydroxypropane sulfonate) Acid
  • two or more of these sulfo group-containing buffering agents may be used in combination, but the amount of the buffering agent added when mixing the buffering agent with serum or plasma is such that serum or plasma lOOmL is added. 10 to 500 mmol is preferred, and 20 to 200 mmol is more preferred.
  • lactate buffer In addition to the buffer with sulfo group, lactate buffer, citrate buffer, acetate buffer, succinate buffer, phthalate buffer, phosphate buffer, triethanolamine buffer, Diethanolamine buffer, lysine buffer, barbitur buffer, imidazole buffer, malate buffer, oxalate buffer, glycine buffer, borate buffer, carbonate buffer, dalysin buffer, Good buffer Agents and the like.
  • Good buffers include, for example, MES (2-morpholinoethanesulfonic acid) buffer, Bis-Tris [bis (2-hydroxyethyl) iminotris (hydroxymethyl) methane] buffer, Tris [tris (hydroxymethyl) aminomethane ] Buffer, ADA [N- (2-acetamido) iminoniacetic acid] buffer, etc. can be used in combination.
  • the standard product of the present invention may contain additives such as an aqueous medium, metal ions, salts, surfactants, preservatives, sugar compounds and the like as necessary.
  • the aqueous medium include deionized water and distilled water.
  • the metal ion include magnesium ion, calcium ion, manganese ion, zinc ion and the like.
  • the salts include sodium salt sodium and potassium salt.
  • the surfactant include nonionic surfactants, cationic surfactants, anionic surfactants, and amphoteric surfactants.
  • antiseptics include antibiotics such as sodium azizuma, streptomycin, gentamicin, Bioace, Procrine 300 (trademark), Proxel GXL (trademark), and the like.
  • sugar compound examples include monosaccharides and disaccharides, with disaccharides being preferred.
  • monosaccharides include glucose, fructose, and fucose.
  • disaccharide examples include saccharose, trehalose, maltose and the like. Two or more monosaccharides and disaccharides can be used in combination.
  • the amount of sugar compound added when sugar compounds are mixed with serum or plasma is preferably 0.5 to 20 g, more preferably 1 to 15 g, relative to serum or plasma lOOmL. 25-12. 5 g is particularly preferred.
  • Reagents for fractionation measurement of components to be measured in lipoproteins include multiple types of lipoproteins that can fractionate specific lipoproteins such as HDL, LDL, VLDL, CM, and remnant lipoproteins by means such as centrifugation. Reagents that measure fractions in specific lipoproteins in the presence of the same reaction solution, so-called homogenous measurement reagents are not particularly limited.
  • Cholesterol measurement reagent in HD L neutral fat in HDL Measuring reagent, cholesterol measuring reagent in LDL, neutral fat measuring reagent in LDL, cholesterol measuring reagent in VLDL, neutral fat measuring reagent in VLDL, cholesterol measuring reagent in CM, neutral fat in CM
  • Examples include a measuring reagent, a cholesterol measuring reagent in remnant lipoprotein, and a neutral fat measuring reagent in remnant lipoprotein.
  • a separate measurement reagent for a component to be measured in a strong lipoprotein for example, not only a reagent prepared in-house but also a commercially available measurement reagent can be used.
  • measuring reagents include, for example, HDL cholesterol measurement reagent Detamina I HDL (Kyowa Medettas), Detamina I HDL (Kyowa Medex), Colles Test N HDL (Daiichi Kagaku) And Detamina L LDL (manufactured by Kyowa Medettas), which is an LDL cholesterol measurement reagent.
  • a sulfo group-containing buffer and serum or plasma are mixed, and the resulting mixture is freeze-dried, and the content of the component to be measured in the freeze-dried product is known.
  • the method is not particularly limited as long as it is a method that uses a component to be measured, and when mixing serum or plasma with a buffer having a sulfo group, the buffer having a sulfo group is an aqueous solution (buffer).
  • the pH of the aqueous solution that can be mixed is selected so that the protein in the serum or plasma does not precipitate after mixing, and pH 6-9 is preferred.
  • lyophilization is used in the usual meaning used in the art, and the sample is frozen, depressurized in the frozen state, and dried by removing moisture from the sample vessel.
  • the lyophilization conditions are not particularly limited, but are usually ⁇ 80 to 20 ° C., preferably ⁇ 80 to 15. It is carried out at a temperature of C at a pressure of 0.6667 to 1333 Pa, preferably 13.3 to 133.3 Pa for 6 to 72 hours, more preferably 12 to 72 hours.
  • the water content of the freeze-dried product is usually 10% by weight or less, preferably 1% by weight or less.
  • Serum or plasma is cooled to 2-8 ° C, and an aqueous solution containing the above-mentioned buffer having a sulfo group is added thereto.
  • an aqueous solution containing the above-mentioned buffer having a sulfo group is added thereto.
  • the pH-adjusted serum or plasma-containing aqueous solution is filtered using a filtration membrane of about 0.2 to 0.4 microns. To do. Dispense the resulting solution into vials for freeze-drying and half-clamp.
  • the solution dispensed in vials is freeze-dried at a low temperature of, for example, -30 ° C or lower, for example, at a vacuum of 133.3 Pa, and finally dried up at 0-20 ° C while gradually warming.
  • a standard product in a lyophilized state Prepare a standard product in a lyophilized state.
  • a lyophilized standard means that the content of the component to be measured in the lipoprotein contained in the manufactured standard is determined using a known amount of the component to be measured. .
  • the measurement components in the lyophilized standard product obtained are standardized by, for example, using multiple homogenous methods such as the recommendation of the US Department of Health's Disease Control and Prevention Center (CDC) recommendation for multiple serum or plasma samples from healthy individuals.
  • CDC US Department of Health's Disease Control and Prevention Center
  • the measured value in the serum or plasma is the CDC recommended method, etc. This can be done by determining the content of the component to be measured in the standard product so that it matches the measured value obtained by the standard method. (Kit for fractional determination of components to be measured in samples containing lipoproteins)
  • the kit for differential quantification of the component to be measured in the sample containing the lipoprotein of the present invention includes the standard product of the present invention and the reagent for differential measurement of the component to be measured in the lipoprotein.
  • the kit of the present invention is used, fractional measurement of components to be measured in a sample containing lipoprotein can be easily performed.
  • a component for measurement of a component to be measured in a lipoprotein is used to determine the component to be measured in a sample.
  • a step of measuring (2) a step of preparing a calibration curve using the standard product in which the amount of the component to be measured according to the present invention is priced and the fractionation measuring reagent, and (3) the step of (1) above. From the measured values obtained and the calibration curve created in step (2), the concentration of the component to be measured in the sample is calculated.
  • fractional measurement step of the component to be measured in the above step (1) there is no particular limitation as long as it is a quantification method having a process for producing a specific step such as HDL, LDL, VLDL, CM, and remnant lipoprotein.
  • a method of fractionating specific components of lipoproteins in a state where multiple types of lipoproteins coexist in the same reaction solution without fractionating lipoproteins by means such as centrifugation, so-called homogenous measurement method is used.
  • the method for measuring the components to be measured for lipoproteins in a specimen by the homogenous method is that the lipoproteins in the specimen are not separated physically, and in the presence of multiple types of lipoproteins, Means a method of measuring the components of
  • the standard product of the present invention and the kit of the present invention are suitably used in a measurement method using an automatic analyzer in which a standard product and a specimen are automatically weighed and measured.
  • Azizuna sodium manufactured by Wako Pure Chemical Industries, Ltd.
  • Bjarbin product name, CV Y-2YT, manufactured by Sato Ampoule Co., Ltd.
  • freeze dryer FDM 05S, manufactured by Sanpec Co., Ltd.
  • Example 1 Each of the freeze-dried products prepared in Example 1 was dissolved in 0.5 mL of purified water, and the concentration of HDL cholesterol in the sample was determined as a commercially available homogenous HDL measuring reagent, Detaminaichi HDL (manufactured by Kyowa Medettas; frozen) Measurement was performed with an automatic analyzer (Hitachi 7170) using a dry reagent product (denoted as reagent DH) and Detamina I L HDL (manufactured by Kyowa Medettas; liquid reagent product (denoted as reagent LH)). Table 1 shows the measurement results.
  • a commercially available homogenous HDL measuring reagent Detaminaichi HDL (manufactured by Kyowa Medettas; frozen) Measurement was performed with an automatic analyzer (Hitachi 7170) using a dry reagent product (denoted as reagent DH) and Detamina I L HDL (man
  • each freeze-dried product was stored for 1 week in a heated state at 40 ° C.
  • Two reagent products for HDL cholesterol measurement used in Example 2 and Correstest N HDL [Daiichi Chemical Co., Ltd .; liquid reagent product (denoted as reagent NH)] and LDL cholesterol measurement reagent Detamina L LDL [ Using a liquid reagent product (reagent LL)] manufactured by Kyowa Medex Co., Ltd.], the measured values before and after storage were compared.
  • the results are shown in Table 2.
  • the numerical values in Table 2 show the ratio of the measured values before and after storage, that is, the measured cholesterol value in the standard product (control product) after storage Z) the measured cholesterol value in the standard product (control product) before storage.
  • a standard product used for differential quantification of components to be measured in lipoprotein, useful for diagnosis of lifestyle-related diseases such as arteriosclerosis a method for producing the standard product, A kit for fractional quantification of a component to be measured and a method for fractional quantification of the component to be measured in a specimen are provided.

Abstract

L’invention concerne un article standard lyophilisé pour utilisation dans la détermination fractionnelle d’un composant à mesurer de lipoprotéine produit par mélange d’un agent tampon sulfoné avec un sérum ou du plasma, lyophilisation dudit article et évaluation de la teneur de composant à mesurer dans le produit de lyophilisation à l’aide d’un composant à mesurer dont la quantité est connue. On trace une courbe analytique en utilisant cet article standard et un réactif pour mesure fractionnelle de composant à mesurer contenu dans une lipoprotéine. On mesure un composant à mesurer d’un analyte à l'aide d'un réactif pour mesure fractionnelle, et l’on calcule la concentration de composant à mesurer de l’analyte à partir de la mesure ainsi obtenue et de la courbe analytique ci-dessus. Ainsi, l’invention concerne un article standard pour détermination d’un composant à mesurer de lipoprotéine assurant une manipulation facile et, même après un stockage sur une longue période, fournissant des valeurs de mesure précises, de même qu’un procédé de fabrication de l’article standard. De plus, elle porte sur un procédé de détermination fractionnelle d’un composant à mesurer d’un analyte contenant une lipoprotéine, employant l’article standard ci-dessus.
PCT/JP2005/020845 2004-11-17 2005-11-14 Article standard pour détermination fractionnelle de composant de lipoprotéine WO2006054519A1 (fr)

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JP2006545011A JP4704355B2 (ja) 2004-11-17 2005-11-14 リポ蛋白中成分の分別定量用標準品

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JP2004-333770 2004-11-17
JP2004333770 2004-11-17

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010156681A (ja) * 2008-12-02 2010-07-15 Kyowa Medex Co Ltd リポ蛋白中成分の分別定量用標準品の製造方法
CN103063829A (zh) * 2012-12-21 2013-04-24 杭州茂天赛科技有限公司 一种冻存液
WO2015076285A1 (fr) * 2013-11-21 2015-05-28 協和メデックス株式会社 Inhibiteur de dénaturation et procédé pour inhiber la dénaturation par lyophilisation de lipoprotéines basse densité contenues dans un sérum sanguin ou un plasma sanguin
JP2015194376A (ja) * 2014-03-31 2015-11-05 シーシーアイ株式会社 中性脂肪濃度測定用の標準物質組成物およびその製造方法
US11604026B2 (en) 2019-03-14 2023-03-14 Terumo Bct Biotechnologies, Llc Lyophilization loading tray assembly and system
US11634257B2 (en) 2017-10-09 2023-04-25 Terumo Bct Biotechnologies, Llc Lyophilization container and method of using same

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JPS58102159A (ja) * 1981-11-30 1983-06-17 テクニコン、インストルメンツ、コーポレーション 安定な基本血清組成物の製造方法
JPS6038657A (ja) * 1983-07-08 1985-02-28 ベーリンガー・マンハイム・ゲゼルシヤフト・ミツト・ベシユレンクテル・ハフツング 貯蔵安定なユニバーサル対照血清
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Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2010156681A (ja) * 2008-12-02 2010-07-15 Kyowa Medex Co Ltd リポ蛋白中成分の分別定量用標準品の製造方法
CN103063829A (zh) * 2012-12-21 2013-04-24 杭州茂天赛科技有限公司 一种冻存液
WO2015076285A1 (fr) * 2013-11-21 2015-05-28 協和メデックス株式会社 Inhibiteur de dénaturation et procédé pour inhiber la dénaturation par lyophilisation de lipoprotéines basse densité contenues dans un sérum sanguin ou un plasma sanguin
JPWO2015076285A1 (ja) * 2013-11-21 2017-03-16 協和メデックス株式会社 血清又は血漿中の低密度リポ蛋白の凍結乾燥による変性抑制剤及び変性抑制方法
TWI651100B (zh) * 2013-11-21 2019-02-21 協和梅德庫斯股份有限公司 甲硫胺酸之用途、血清或血漿中低密度脂蛋白因冷凍乾燥所致的變性之抑制方法、血清或血漿中低密度脂蛋白中之成分定量用標準品、其製造方法與應用
JP2015194376A (ja) * 2014-03-31 2015-11-05 シーシーアイ株式会社 中性脂肪濃度測定用の標準物質組成物およびその製造方法
US11634257B2 (en) 2017-10-09 2023-04-25 Terumo Bct Biotechnologies, Llc Lyophilization container and method of using same
US11609043B2 (en) 2019-03-14 2023-03-21 Terumo Bct Biotechnologies, Llc Lyophilization container fill fixture, system and method of use
US11609042B2 (en) 2019-03-14 2023-03-21 Terumo Bct Biotechnologies, Llc Multi-part lyophilization container and method of use
US11604026B2 (en) 2019-03-14 2023-03-14 Terumo Bct Biotechnologies, Llc Lyophilization loading tray assembly and system
US11740019B2 (en) 2019-03-14 2023-08-29 Terumo Bct Biotechnologies, Llc Lyophilization loading tray assembly and system
US11747082B2 (en) 2019-03-14 2023-09-05 Terumo Bct Biotechnologies, Llc Multi-part lyophilization container and method of use
US11815311B2 (en) 2019-03-14 2023-11-14 Terumo Bct Biotechnologies, Llc Lyophilization container fill fixture, system and method of use

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JP4704355B2 (ja) 2011-06-15

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