Method for assaying glycated albumin

• US 7,871,789 B2
• Filed: 10/27/2006
• Issued: 01/18/2011
• Est. Priority Date: 10/27/2005
• Status: Active Grant

• Current Assignee: ARKRAY, Inc
• Original Assignee: ARKRAY, Inc
• Inventors: Imamura, Norio, Yonehara, Satoshi
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US12/084,003
• Publication Number:
  US 20090142787A1
• Time in Patent Office: 1,544 Days
• Field of Search: 435/14, 435/23
• US Class Current: 435/23
• CPC Class Codes: C12Q 1/37 involving peptidase or proteinase
• CPC Class Codes: G01N 2333/76 Assays involving albumins other than in routine use for blocking surfaces or for anchoring haptens during immunisation
• CPC Class Codes: G01N 2400/00 Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
• CPC Class Codes: G01N 33/68 involving proteins, peptides or amino acids involving lipoproteins G01N33/92

Abstract

An albumin denaturing agent for digesting an albumin by a protease efficiently is provided. The albumin denaturing agent contains quaternary ammonium having a hydrocarbon group with a carbon number of 12 or more, or a salt of the quaternary ammonium. The albumin in a sample is digested by the protease in the presence of the albumin denaturing agent, a glycated part of the thus obtained albumin digestion product and a FAOD effect a reaction, and a redox reaction between the glycated part and the FAOD is measured, thereby determining a ratio (GA (%)) of the glycated albumin of the glycated albumin with respect to the albumin.

Claims

1. A method for assaying glycated albumin, comprising:

adding a metalloprotease into a sample containing the glycated albumin so as to digest the glycated albumin with the metalloprotease;

adding quaternary ammonium having a hydrocarbon group with a carbon number of 14 or more, or a salt of the quaternary ammonium before, at the same time of or after the adding of the metalloprotease, wherein a resultant concentration of the metalloprotease ranges from 1 KU/L to 1,600 KU/L,

adding a fructosyl amino acid oxidase (FAOD) to the sample so that a glycated part of the digestion product of the glycated albumin reacts with the FAOD, wherein the FAOD is added before, at the same time or after the adding of the metalloprotease;

measuring an amount of a redox reaction product generated by a redox reaction between the glycated part of the digestion product of the glycated albumin and the FAOD; and

correlating the measured amount of the redox reaction product with an amount of the glycated albumin in the sample.

3. The method of claim 1, wherein the redox reaction product measured is hydrogen peroxide, wherein measuring the amount of the redox reaction product includes adding a peroxidase and a chromogenic substrate to the sample so that the hydrogen peroxide reacts with the chromogenic substrate, and measuring a chromogenic level of the chromogenic substrate, and wherein correlating the measured amount of the redox reaction product with an amount of the glycated albumin includes calculating the amount of the glycated albumin based on the measured chromogenic level of the chromogenic substrate.

2. A method for assaying glycated albumin, comprising:

adding a metalloprotease into a sample containing the glycated albumin so as to digest the glycated albumin with the metalloprotease;

adding quaternary ammonium having a hydrocarbon group with a carbon number of 14 or more, or a salt of the quaternary ammonium before, at the same time of or after the adding of the metalloprotease, wherein an adding amount of the metalloprotease ranges from 1 KU to 60 KU in 1 ml of the sample;

adding a fructosyl amino acid oxidase (FAOD) to the sample so that a glycated part of the digestion product of the glycated albumin reacts with the FAOD, wherein the FAOD is added before, at the same time or after the adding of the metalloprotease;

measuring an amount of a redox reaction product generated by a redox reaction between the glycated part of the digestion product of the glycated albumin and the FAOD; and

correlating the measured amount of the redox reaction product with an amount of the glycated albumin in the sample.

4. The method of claim 2, wherein the redox reaction product measured is hydrogen peroxide, wherein measuring the amount of the redox reaction product includes adding a peroxidase and a chromogenic substrate to the sample so that the hydrogen peroxide reacts with the chromogenic substrate, and measuring a chromogenic level of the chromogenic substrate, and wherein correlating the measured amount of the redox reaction product with an amount of the glycated albumin includes calculating the amount of the glycated albumin based on the measured chromogenic level of the chromogenic substrate.