Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
  • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
  • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
  • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase

Definitions

• the present invention relates to a test sample for measuring the activity of glutamyl transpeptidase. More specifically, the present invention is characterized in that it contains r-glutamyl-4-nitronitronide and cyclodextrin. The present invention relates to a reagent for measuring the activity of r-glutamyl transpeptidase.
• the present inventors have conducted various studies on a method for measuring the enzymatic activity of O-OTP using Glut-4-NA as a substrate, and found that the reaction system had a cyclodextrin. It has been found that the addition of C can significantly increase the solubility of Olu-4-NA and also inhibit the non-excitable degradation of C-lu-4-NA in water or buffer solutions. did. The present invention has been made based on such knowledge. Is
• r-glutamyl-4- 4-trao-lide (substrate) is used as a reagent for measuring the enzymatic activity of r-ciTP, to which cyclodextrin is added.
• This is a feature, and its purpose is to use r-daltamyl- as a substrate.
• Another object of the present invention is to provide a test method capable of remarkably and accurately measuring the activity of daltamyl transdibutidase as compared with the prior art.
• Glu-4-NA used as a substrate has a solubility in water or buffer of up to 4 mM at most, as described above.
• a high solubility of 10 m (4 times higher) can be obtained by adding, for example, six dextrin 0.3 (wv).
• cyclodextrin has the effect of stabilizing C-lu-4-1NA in addition to the above-mentioned increase in solubility of Olu-14-NA. That is, when the aqueous solution or buffer solution (4 mLi) of Glu-4 -'-: ⁇ is left at room temperature, the decomposition amount has already reached about 27 AM in 24 hours and about 5 in 48 hours. It shows a value of about 80 M at 72 hours at 72 hours and about 108 # ⁇ at 90 hours, but this decomposition is due to this decomposition.] 4-nitronitrolin produced is r -After 24 hours, to prevent the catalysis of, already? "-It was found that the enzyme activity of -CT?
• Cost and water solubility are recommended as the preferred modes of use for ⁇ -cyclodextrin.
• An advantage to be noted with respect to the reagent of the present invention is that it can be provided as a lyophilized product. That is, conventionally, the r-GTP enzyme activity assay containing G1U-4-NA as a main component has been described above.3) Since Gin-4-NA is hardly soluble, various solubilization means are used. Finally, it was only possible to obtain a concentration of about 4 m, and it was almost impossible to produce a freeze-dried product as a prototype, but according to the present invention, By using in combination with cyclodextrin], it has become possible to provide freeze-dried products. That is, (r-GTP can be measured by a simple operation using a freeze-dried mixed solution of Jlu-4-NA and cyclodextrin. This will provide significant advantages in making the task of measuring 7 "-GTP significantly easier.
• Buffer solution Glycyl Glycine 6.2 and diethanolamine 10.5 ⁇ are dissolved in about 900% of distilled water, and the pH is adjusted using saline. After adding & 2 (301C), distilled water was added to give 100 ⁇ .
• Substrate solution Dissolve L-7 * -Gnoretamyl-4-nitrofuranilide 4. ⁇ and cyclodextrin 2.7 ⁇ with the above buffer solution.
• Measurement solution 300 ml of the buffer solution of the above H) and the substrates v, The solution is mixed with to obtain a solution.
• the amount of increase in absorbance per minute for 1 minute is defined as X.
• Unit (activity) 1 00 ⁇ ⁇ 2 ⁇ 2 ⁇
• the resulting lyophilized product is dissolved in water or buffer solution (3) to give a 16 mM Glu-4-NA solution.
• the above freeze-dried product is much more stable than the solid state of Glu-4-HA alone]], and this product can be stored in a stable state.
• the reagent for measuring the activity of 7 * -glutamyl transpeptidase of the present invention is r-glutamyl transpeptidase (r
• the reagent for measuring r-GTP activity of the present invention is usefully used for medical diagnosis and also for enzymatic chemistry research, its industrial applicability is extremely large. You.

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• Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Life Sciences & Earth Sciences (AREA)
• Zoology (AREA)
• Wood Science & Technology (AREA)
• Proteomics, Peptides & Aminoacids (AREA)
• Health & Medical Sciences (AREA)
• Engineering & Computer Science (AREA)
• Microbiology (AREA)
• Immunology (AREA)
• Physics & Mathematics (AREA)
• Molecular Biology (AREA)
• Biotechnology (AREA)
• Biophysics (AREA)
• Analytical Chemistry (AREA)
• Biochemistry (AREA)
• Bioinformatics & Cheminformatics (AREA)
• General Engineering & Computer Science (AREA)
• General Health & Medical Sciences (AREA)
• Genetics & Genomics (AREA)
• Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

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• 1980
  • 1980-10-28 JP JP55150240A patent/JPS5774099A/ja active Granted
• 1981
  • 1981-10-28 WO PCT/JP1981/000303 patent/WO1982001564A1/ja not_active Ceased
  • 1981-10-28 DE DE3152458T patent/DE3152458C2/de not_active Expired - Lifetime

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