公开(公告)号：US20150225768A1

公开(公告)日：2015-08-13

申请号：US14429784

申请日：2013-09-19

Applicant: DKK-TOA CORPORATION ,  HIROSHIMA UNIVERSITY

Inventor： Satoshi Yawata ,  Hiromitsu Hachiya ,  Akio Kuroda ,  Kenichi Noda

IPC: C12Q1/66 ,  C12Q1/00

CPC classification number: C12Q1/66 ,  C12Q1/008 ,  G01N21/76 ,  G01N33/579 ,  G01N2400/24 ,  G01N2400/50

Abstract: A quantification method includes a calibration curve preparing step to measure a standard solution, which has been prepared by adding sodium ions so that a sodium ion content of the standard solution is equaled to a sodium ion content of a sample to be measured with a method employing a reaction that activates a limulus reagent and/or a biochemical luminescent reaction caused by ATP, luciferin, and luciferase, and to prepare a calibration curve that represents a relation between a measurement value and an amount of a component to be measured; a sample measuring step to measure the sample to be measured with a method being the same as that used in the calibration curve preparing step; and a quantifying step to find, by using the calibration curve, an amount of the component to be measured in the sample to be measured from a measurement value in the sample measuring step.

Abstract translation: 定量方法包括：通过添加钠离子制备标准溶液的标准溶液的校准曲线制备步骤，使标准溶液的钠离子含量与待测样品的钠离子含量相等， 激活鲎试剂和/或由ATP，荧光素和荧光素酶引起的生化发光反应的反应，并制备表示测量值与待测组分量之间的关系的校准曲线; 样品测量步骤，用与校准曲线制备步骤中使用的方法相同的方法测量待测样品; 以及量化步骤，通过使用校准曲线，从样品测量步骤中的测量值来求出待测样品中待测量的成分的量。

公开(公告)号：US10851141B2

公开(公告)日：2020-12-01

申请号：US16311301

申请日：2017-06-19

Applicant: DKK-TOA Corporation

Inventor： Kenichi Noda ,  Satoshi Yawata ,  Ai Shimomura

IPC: C12N15/10 ,  C12N15/09 ,  C12P21/02 ,  C07H21/04 ,  C07K14/435 ,  C12N9/02 ,  C12N15/63

Abstract: The present invention provides a mutant beetle luciferase and the like, having mutation in which the amino acid corresponding to valine at position 288 in the amino acid sequence of wild-type Photinus pyralis luciferase is isoleucine, leucine or phenylalanine, mutation in which the amino acid corresponding to leucine at position 376 in the aforementioned sequence is proline, mutation in which the amino acid corresponding to glutamic acid at position 455 in the aforementioned sequence is valine, alanine, serine, leucine, isoleucine or phenylalanine, or mutation in which the amino acid corresponding to glutamic acid at position 488 in the aforementioned sequence is valine, alanine, serine, leucine, isoleucine or phenylalanine, in the amino acid sequence encoding a wild-type beetle luciferase, and characterized in that a luminescence intensity due to a luciferin-luciferase luminescence reaction in a 0.9% by mass NaCl solution is 50% or more of that in a NaCl-free solution.

公开(公告)号：US20190352350A1

公开(公告)日：2019-11-21

申请号：US16311301

申请日：2017-06-19

Applicant: DKK-TOA Corporation

Inventor： Kenichi Noda ,  Satoshi Yawata ,  Ai Shimomura

IPC: C07K14/435 ,  C12P21/02 ,  C12N9/02 ,  C12N15/63

Abstract: The present invention provides a mutant beetle luciferase and the like, having mutation in which the amino acid corresponding to valine at position 288 in the amino acid sequence of wild-type Photinus pyralis luciferase is isoleucine, leucine or phenylalanine, mutation in which the amino acid corresponding to leucine at position 376 in the aforementioned sequence is proline, mutation in which the amino acid corresponding to glutamic acid at position 455 in the aforementioned sequence is valine, alanine, serine, leucine, isoleucine or phenylalanine, or mutation in which the amino acid corresponding to glutamic acid at position 488 in the aforementioned sequence is valine, alanine, serine, leucine, isoleucine or phenylalanine, in the amino acid sequence encoding a wild-type beetle luciferase, and characterized in that a luminescence intensity due to a luciferin-luciferase luminescence reaction in a 0.9% by mass NaCl solution is 50% or more of that in a NaCl-free solution.

公开(公告)号：US08507216B2

公开(公告)日：2013-08-13

申请号：US13760524

申请日：2013-02-06

Applicant: Hiroshima University

Inventor： Akio Kuroda ,  Kenichi Noda

IPC: C12Q1/56

CPC classification number: G01N21/763 ,  C12Q1/66 ,  G01N33/56911 ,  G01N33/579

Abstract: The present invention provides a method comprising allowing a reaction of a sample, a reagent containing Factor C, which can be activated by binding with endotoxin, and a synthetic luminescent substrate comprising a luminescent substrate bound to a peptide, for release of the luminescent substrate from the synthetic luminescent substrate, allowing a luminescent enzyme to act on the luminescent substrate released in the luminescent substrate release step, for measurement of the luminescence intensity, and quantifying the level of endotoxin in the sample based on a measured value obtained in the luminescence measuring step, the method enabling endotoxin to be simply and quickly measured at a level that cannot be detected in conventional methods for endotoxin measurement, without use of any dedicated measuring device.

Abstract translation: 本发明提供了一种方法，其包括使样品，可通过与内毒素结合而活化的因子C的试剂与包含与肽结合的发光底物的合成发光底物反应，从而将发光底物从 所述合成发光基板允许发光酶作用于在所述发光底物释放步骤中释放的所述发光基板，用于测量所述发光强度，并且基于在所述发光测量步骤中获得的测量值来定量所述样品中的内毒素水平 ，在不使用任何专用测量装置的情况下，能够以常规的内毒素测量方法无法检测的水平简单且快速地测量内毒素的方法。