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Method and kit for scaling investigating angiotensin aminopherase

An angiotensin, converting enzyme technology, applied in biological testing, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of complex experimental steps, inappropriate clinical biochemical testing, etc., and achieve the effect of improving test accuracy

Inactive Publication Date: 2005-11-09
ZHEJIANG YAKE SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Although Hurst's method can quantitatively measure ACE activity, the experimental steps are complicated and not suitable for clinical biochemical tests

Method used

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  • Method and kit for scaling investigating angiotensin aminopherase
  • Method and kit for scaling investigating angiotensin aminopherase
  • Method and kit for scaling investigating angiotensin aminopherase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0012] 1. Determination of ACE standard and its activity

[0013] 1) ACE standard composition

[0014] 0-200U / L human gene recombinant ACE

[0015] 2) Determination of the activity of ACE standard

[0016] Referring to Hurst's method, add 0.1ml distilled water and 0.1ml ACE standard (0-200U / L) to a test tube containing 0.2ml incubation buffer (200mM boric acid, 2M sodium chloride pH 8.3), and incubate at 37°C for 5 minutes. The reaction was initiated by adding 0.1 ml of 20 mM HHL substrate solution for 15 minutes. Add 0.5ml of 1M hydrochloric acid (HCl) solution to terminate the reaction, and add 0.5ml of 1M sodium hydroxide (NaOH) after 30 seconds to neutralize the reaction. Add 2ml of 200mM potassium phosphate diluent pH 8.3, followed by 1.5ml of chromogenic solution (3% cyanuric acid chloride dissolved in 1,4-dioxane), mix well, let stand for 5 minutes, mix well again, and centrifuge at 3000 rpm 10 minutes. The supernatant was taken to quantitatively measure the absorb...

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Abstract

Buttery's(1993) velocity of FAPGG method for measuring the activity of hypertension transformation enzyme on the clinical chemistry examination is applied widely. However, Buttery's method lacks a generally accepted standard method to mark the samples, thus the test accuracy often descends. Moreover, different laboratories apply dissimilar biochemical analysis instruments, and the normal reference data scope for hypertension transformation enzyme is usually confused and hardly compared with others. The invention uses the Hurst's method(1981) to examine the activity of the standard hypertension transformation enzyme. Then, following the standard activity, it applies the Buttery's method to measure the activity of hypertension transformation enzyme in living creature samples quantificationally. The invention has solved or reduced the blemishes of the Buttery's method. The invention also involves a reagent box used for implementing the method mentioned above.

Description

technical field [0001] The invention relates to a technical scheme for measuring the activity of angiotensin converting enzyme in biological samples and a reagent produced by the scheme. In particular, it relates to a technical scheme for standardizing the FAPGG rate method for quantitative determination of angiotensin-converting enzyme activity. Background technique [0002] Buttery (1993) used angiotensin converting enzyme (Angiotensin Converting Enzyme, ACE) to enzymolyze phenylalanyl glycylglycine (furylacryloylphenylalanylglycylglycine, FAPGG) to generate phenylalanyl (furylacryloylphenylalanine, FAP) and diglycine Peptide (glycylglycine, GG). The maximum absorption peak of FAPGG is at 340nm, and the enzymatic reaction causes the absorbance at 340nm to decrease. The activity of ACE can be calculated by continuously monitoring the rate of decrease of the absorbance of FAPGG at 340nm. [0003] One unit of ACE activity is defined as the amount of ACE enzyme required to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/33
Inventor 蔡枫
Owner ZHEJIANG YAKE SCI & TECH
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