Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase
Patent
1999-08-13
2000-10-03
Leary, Louise N.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving hydrolase
435 24, 435 4, 4352547, 4352565, 4352561, 435929, C12Q 137, C12Q 100, C12N 114
Patent
active
061271381
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a method of measuring a glycated protein by using fructosyl amino acid oxidase (hereinafter, referred to as "FAOD"). More particularly, the present invention relates to a method which is capable of measuring a glycated protein with greater accuracy and high sensitivity, and is easily applicable to clinical examination (or clinical diagnostic test), etc., and also relates to a protease which is suitably usable for such a measurement method.
BACKGROUND ART
A glycated protein is a substance which is produced by the non-enzymatic and irreversible binding of the amino group of an amino acid constituting a protein, with the aldehyde group of a reducing sugar such as aldose. Such a non-enzymatic and irreversible binding reaction is also called "Amadori rearrangement," and therefore the above-mentioned glycated protein may also be called "Amadori compound" in some cases.
The rate of the formation of the glycated protein generally depends on the concentration of the protein and the reducing sugar as raw materials for providing the above glycated protein, the time period of the contact between these raw materials, and the temperature at the time of the glycation reaction. As a matter of course, as the amount of the above protein and reducing sugar is increased, as the time period of the contact therebetween is increased, or as the temperature becomes higher (within a range such that the protein is not denatured), the rate of the formation of the glycated protein as a reaction product is increased, and the amount of the reaction product is also increased.
On the other hand, in a living organism (or "in vivo"), since the concentration of the glycated protein is changed depending on the half life of the protein as the raw material for the above glycation reaction, various kinds of information on the living organism can be obtained by measuring the concentration of the glycated protein.
Among the above-mentioned glycated proteins, for example, a fructosylamine derivative produced by the glycation of hemoglobin in blood is called "glycohemoglobin", one produced by the glycation of albumin is called "glycoalbumin", and a derivative (having a reducing ability) produced by the glycation of protein in blood is called "fructosamine".
Since the concentration of these glycated protein derivatives in blood reflects the average concentration of blood sugar in a living organism for a certain period of time in the past, the measured value of the concentration of the above glycated protein derivative in blood may be a significant indicator of the diagnosis of the symptom of diabetes and of the monitor or control of such a symptom. Accordingly, also from a clinical viewpoint, it is very useful to establish a method of measuring the concentration of the glycated protein in blood.
Heretofore, it has been known that a glycated protein in a sample (or specimen) can be measured, e.g., by causing an oxidoreductase to act on the glycated protein and measuring the amount of the oxygen consumed in this reaction or the amount of the product (such as hydrogen peroxide) based on the action of the oxidoreductase (e.g., Japanese Patent Publication (JP-B; "Kokoku") Hei-5-33997 (i.e., 33997/1993), JP-B Hei-6-65300, Japanese Laid-Open Patent Applications (JP-A; "Kokai") Hei-2-195900, JP-A Hei-3-155780, JP-A Hei-4-4874, JP-A Hei-5-192193, JP-A Hei-6-46846, JP-A Hei-7-289253, JP-A Hei-8-154672 and JP-A Hei-8-336386 may be referred to).
In addition, there is known a method of measuring a glycated protein for the purpose of the diagnosis of diabetes (JP-A Hei-2-195899, JP-A Hei-2-195900, JP-A Hei-5-192193 corr. to European Publication EP 0526150A, JP-A Hei-6-46846 corr. to EP 0576838A, JP-A Hei-7-289253, JP-A Hei-8-154672 and JP-A Hei-8-336386 may be referred to).
In general, an enzymatic reaction using a glycated protein as a substrate is represented by the following formula. --CO--CHO+R.sup.2 --NH.sub.2 +H.sub.2 O.sub.2 R.sup.2 represents a residue of an amino acid, protein or peptide.)
As an enzyme
REFERENCES:
patent: 5712138 (1998-01-01), Kato et al.
Luisetti M., et al.; "Some Properties of the Alkaline Proteinase from Aspergillus Melleus"; Int. J. Tiss. Reac. XIII(4), pp. 187-192 (1991). Month not available.
Fukuya Hiroshi
Ishimaru Kaori
Okamura Akihiko
Ooishi Katsutaka
Kyoto Dai-ichi Kagaku Co. Ltd.
Leary Louise N.
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