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    Highly sensitive enzyme assay method
    41.
    发明授权
    Highly sensitive enzyme assay method 失效
    高灵敏度酶法测定

    公开(公告)号：US4693971A

    公开(公告)日：1987-09-15

    申请号：US575292

    申请日：1984-01-30

    申请人： Hideo Misaki

    发明人： Hideo Misaki

    IPC分类号： C12Q1/00 ,  C12Q1/26 ,  C12Q1/50 ,  C12Q1/32 ,  C12Q1/44

    CPC分类号： C12Q1/26 ,  C12Q1/008

    摘要： A highly sensitive quantitative assay method for any one component which is L-glycero-3-phosphate (G3P), dihydroxyacetone-3-phosphate (DHAP), nicotinamide adenine dinucleotide (NAD) or reduced NAD, in a specimen to be assayed, comprising causing this component in the specimen to take part in the cycling reaction ##STR1## wherein GPO is glycerophosphate oxidase and GPDH is glycerophosphate dehydrogenase, and measuring a detectable change in the reaction system. There is thus provided a novel G3P-GHAP cycling reaction using GPO, which consumes O.sub.2 and generates H.sub.2 O.sub.2 and DHAP, with a substrate of G3P, and furthermore GPDH which consumes reduced NAD and generates NAD and G3P, with a substrate of DHAP. Examples of specimens are specimens which contain any one of G3P, DHAP, NAD or reduced NAD, or which liberate or generate such a component. By proceeding at a rate of more than ten cycles per minute and measuring the amount of a detectable change in the reaction, the component in a specimen can easily and sensitively be detected with good accuracy.

    摘要翻译： 对待测试样品中任何一种成分为L-甘油-3-磷酸（G3P），二羟基丙酮-3-磷酸（DHAP），烟酰胺腺嘌呤二核苷酸（NAD）或还原型NAD）的高灵敏度定量测定方法，包括 使样品中的该组分参与循环反应，其中GPO是甘油磷酸氧化酶，GPDH是甘油磷酸脱氢酶，并测量反应体系中可检测的变化。 因此，提供了使用GPO的新颖的G3P-GHAP循环反应，其使用G3P的底物消耗O 2并产生H 2 O 2和DHAP，此外还使用消耗NAD并产生NAD和G3P的GPDH与DHAP的底物。 样品的实例是含有G3P，DHAP，NAD或还原型NAD中的任何一种或释放或产生这种组分的样品。 通过以每分钟超过10个循环的速率进行测量并测量反应中可检测的变化量，可以以高精度容易且灵敏地检测样品中的组分。

    Element for immunoassay
    42.
    发明授权
    Element for immunoassay 失效
    免疫测定的要素

    公开(公告)号：US4495151A

    公开(公告)日：1985-01-22

    申请号：US394473

    申请日：1982-07-01

    申请人： Kunio Ohyama ,  Nobuaki Nakagawa ,  Susumu Watanabe

    发明人： Kunio Ohyama ,  Nobuaki Nakagawa ,  Susumu Watanabe

    IPC分类号： G01N33/543 ,  G01N33/54

    CPC分类号： G01N33/54366 ,  Y10S436/81

    摘要： Apparatus for performing an immunoassay, comprises in combination a reaction tube closed at its bottom, and in the reaction tube a molded element having on its surface material having specific binding ability for a component in a liquid specimen to be assayed. The shape of the surface of the element adjacent the interior surface of the tube is the complement of the shape of the interior surface of the tube. A spacer rib is provided on the element, in contact with the interior surface of the tube. This rib holds the element in such position in the tube that the surfaces of complementary shape of the element and tube are shaped from each other a small but constant distance. A thin layer of constant thickness of a liquid specimen for immunoassay is thus receivable between the complementary surfaces. This arrangement increases the contact surface of the liquid medium in the reaction tube by the element, by establishing a homogeneous uniform layer of liquid medium. This avoids the need for stirring and shaking and reduces the required volume of the specimen to be assayed.

    摘要翻译： 用于进行免疫测定的装置包括在其底部封闭的反应管，反应管中的模制元件在其表面上具有对要测定的液体样品中的组分的特异性结合能力的材料。 邻近管内表面的元件表面的形状是管的内表面形状的互补。 间隔肋设置在元件上，与管的内表面接触。 该肋将元件保持在管中的这种位置，元件和管的互补形状的表面彼此成一个小但恒定的距离。 因此，可以在互补表面之间接收用于免疫测定的液体样本的恒定厚度的薄层。 这种布置通过构成均匀均匀的液体介质层，增加了反应管中液体介质的接触表面。 这避免了搅拌和摇动的需要，并且减少要测定的样品的所需体积。

    Assay method for amylase activity and method of producing maltose dehydrogenase for use therein
    43.
    发明授权
    Assay method for amylase activity and method of producing maltose dehydrogenase for use therein 失效
    淀粉酶活性的测定方法和生产用于其中的麦芽糖脱氢酶的方法

    公开(公告)号：US4427771A

    公开(公告)日：1984-01-24

    申请号：US311263

    申请日：1981-10-14

    申请人： Hideo Misaki ,  Eiji Muramatsu ,  Hidehiko Ishikawa ,  Kazuo Matsuura

    发明人： Hideo Misaki ,  Eiji Muramatsu ,  Hidehiko Ishikawa ,  Kazuo Matsuura

    IPC分类号： C12N9/04 ,  C12Q1/40 ,  C12N1/20 ,  C12Q1/32 ,  C12R1/11

    CPC分类号： C12N9/0006 ,  C12Q1/40 ,  G01N2400/18 ,  Y10S435/837

    摘要： An assay method for amylase activity in a biological specimen such as serum, saliva or urine. The enzyme amylase in the specimen is used to decompose a substrate which is a glucose polymer having a modified reducing terminal glucose residue or a cyclic glucose polymer. A component of the decomposed substrate is measured as an indication of amylase activity in the specimen. The residue may be amylose, amylopectin, starch, starch hydrolyzate, an etherified reducing terminal, an esterified reducing terminal, gluconolactone or a gluconic acid residue or its derivative. Decomposed substrate assay may be effected by contacting the same with maltose dehydrogenase and NAD or NADP, whereupon the assay is performed by measuring the amount of reduced NAD or reduced NADP, by reacting the same with reduced-form hydrogen transport colorimetric reaction reagent. This reagent may be a tetrazolium salt and diaphorase, or tetrazolium salt and phenazinemethosulfate. To remove pre-existing glucose and maltose present in the specimen, the specimen may be pretreated with alpha-glucosidase or kinase in the presence of Mg.sup.++ and ATP, the kinase being for example hexokinase. The preferred maltose dehydrogenase is produced by culturing Bacillus megaterium B-0779 FERM-P No. 5662.

    摘要翻译： 生物样品如血清，唾液或尿液中淀粉酶活性的测定方法。 试样中的酶淀粉酶用于分解具有修饰的还原性末端葡萄糖残基或环状葡萄糖聚合物的葡萄糖聚合物的底物。 测量分解的底物的组分作为样品中淀粉酶活性的指示。 残基可以是直链淀粉，支链淀粉，淀粉，淀粉水解物，醚化还原末端，酯化还原末端，葡萄糖酸内酯或葡萄糖酸残基或其衍生物。 分解的底物测定可以通过使其与麦芽糖脱氢酶和NAD或NADP接触来实现，因此通过使其与还原形式的氢转运比色反应试剂反应来测量还原的NAD或还原的NADP的量来进行测定。 该试剂可以是四唑盐和心律黄素，或四唑鎓盐和吩嗪甲硫酸盐。 为了除去样品中存在的现有葡萄糖和麦芽糖，可以在Mg ++和ATP存在下用α-葡糖苷酶或激酶预处理样品，所述激酶例如己糖激酶。 优选的麦芽糖脱氢酶是通过培养巨大芽孢杆菌B-0779 FERM-P No.5662产生的。

    Assay method of enzyme activity
    44.
    发明授权
    Assay method of enzyme activity 失效
    酶活性的测定方法

    公开(公告)号：US4384041A

    公开(公告)日：1983-05-17

    申请号：US251784

    申请日：1981-04-07

    申请人： Kunio Matsumoto ,  Toshiharu Noda ,  Tadashiro Fujii ,  Susumu Watanabe ,  Kazuhiko Matsumoto ,  Shigeo Katsuragi

    发明人： Kunio Matsumoto ,  Toshiharu Noda ,  Tadashiro Fujii ,  Susumu Watanabe ,  Kazuhiko Matsumoto ,  Shigeo Katsuragi

    IPC分类号： C07C271/22 ,  C07D209/20 ,  C07D233/64 ,  C07K5/06 ,  C12Q1/37 ,  C12N11/00 ,  C12Q1/26 ,  C12Q1/28 ,  C12Q1/36

    CPC分类号： C07K5/06043 ,  C07C271/22 ,  C07D209/20 ,  C12Q1/37 ,  Y10S435/817

    摘要： Novel amine compounds of the formula ##STR1## wherein R.sub.1 is lower alkyl and R.sub.2 is substituted methyl amino or D-amino acid residue, or a salt thereof. R.sub.1 can typically be isobutyl or methyl; and R.sub.2 can be methylamino of the formula--NH--CH.sub.2 --R.sub.3in which R.sub.3 is organic, more particularly methyl, ethyl, n-propyl, isopropyl, n-butyl, amyl, p-hydroxybenzyl, 3,4-di-hydroxybenzyl, 5-imidazolemethyl, 3-imidazolemethyl or phenyl. R.sub.2 can also be D-amino acid residue of the formula ##STR2## in which one of R.sub.4 and R.sub.5 is carboxyl and the other is organic, more particularly methylthioethyl, isobutyl, methyl or phenyl, and C* is a D-asymmetric carbon atom. The compounds are useful in an assay method for leucine and aminopeptidase which comprises reacting a substrate amide as above, or a salt thereof, with a sample for leucine aminopeptidase assay, incubating the reaction product produced thereby, and measuring the thus-consumed oxygen or liberated hydrogen peroxide.

    摘要翻译： 其中R 1为低级烷基且R 2为取代的甲基氨基或D-氨基酸残基的新颖胺化合物或其盐。 R1通常可以是异丁基或甲基; 并且R 2可以是式-NH-CH 2 -R 3的甲基氨基，其中R 3是有机的，更特别的是甲基，乙基，正丙基，异丙基，正丁基，戊基，对羟基苄基，3,4-二羟基苄基， 5-咪唑甲基，3-咪唑甲基或苯基。 R2也可以是式“IMAGE”的D-氨基酸残基，其中R4和R5之一是羧基，另一个是有机的，更特别是甲硫基乙基，异丁基，甲基或苯基，C *是D-不对称碳原子 。 该化合物可用于亮氨酸和氨基肽酶的测定方法中，其包括如上所述的底物酰胺或其盐与用于亮氨酸氨基肽酶测定的样品反应，孵育由此产生的反应产物，并测量如此消耗的氧气或释放的 过氧化氢。