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Liquid single reagent for detecting alpha-L-fucosidase and preparation method thereof

A fucopyranoside, single-reagent technology, applied in the measurement of color/spectral properties, etc., can solve the problems of ineffective resistance to heparin interference and poor stability.

Inactive Publication Date: 2013-03-13
BEIJING LEADMAN BIOCHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] The first technical problem to be solved by the present invention is to overcome the defects of poor stability and inability to effectively resist heparin interference in the existing liquid single reagent for detecting α-L-fucosidase, and provide a new method for detecting α-L-fucosidase. Liquid single reagent for fucosidase with excellent stability and resistance to interference from heparin in samples

Method used

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  • Liquid single reagent for detecting alpha-L-fucosidase and preparation method thereof
  • Liquid single reagent for detecting alpha-L-fucosidase and preparation method thereof
  • Liquid single reagent for detecting alpha-L-fucosidase and preparation method thereof

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Embodiment 1

[0034] Embodiment 1 Preparation of liquid single reagent of the present invention

[0035] Add glycine-hydrochloric acid buffer solution with a pH value of 3.0, ethylenediaminetetraacetic acid, NaCl, surfactant Tween20, stabilizer methanol and chlorinated p-nitrophenyl α-L-fucopyranoside into distilled water and mix well And fully dissolved in water to obtain liquid single reagent;

[0036] Among them, the amount of each component in 1 liter of liquid single reagent is controlled as 0.02 molar mass of glycine-hydrochloric acid buffer solution with a pH value of 3.0, 0.01 g of ethylenediamine tetraacetic acid, 5 g of NaCl, 0.1 g of surfactant Tween20, and stabilizer 0.1 g of methanol, 0.5 g of chlorinated p-nitrobenzene α-L-fucopyranoside, and the balance is water.

Embodiment 2

[0037] Embodiment 2 Preparation of liquid single reagent of the present invention

[0038] Disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.0, ethylenediaminetetraacetic acid, NaCl, surfactant Triton X-100, stabilizer ethylene glycol and chlorinated p-nitrobenzene α-L-fucus Add pyranoside to distilled water, mix evenly and fully dissolve in water to obtain liquid single reagent;

[0039] Among them, the amount of each component in 1 liter of liquid single reagent is controlled as follows: 0.05 molar mass of disodium hydrogen phosphate-citric acid buffer solution with a pH value of 5.0, 0.5 g of ethylenediamine tetraacetic acid, 10 g of NaCl, and surfactant Triton X -100 10 g, stabilizer ethylene glycol 10 g, p-nitrophenyl chloride α-L-fucopyranoside 2 g, and the balance is water.

Embodiment 3

[0040] Embodiment 3 Preparation of liquid single reagent of the present invention

[0041] Citric acid-sodium citrate buffer at pH 4.0, ethylenediaminetetraacetic acid, NaCl, surfactant Brij 35, stabilizer isopropanol, and p-nitrophenyl α-L-fucopyranoside chloride Add to distilled water, mix well and fully dissolve in water to obtain liquid single reagent;

[0042] Among them, the amount of each component in 1 liter of liquid single reagent is controlled as 0.02 molar mass of citric acid-sodium citrate buffer solution with a pH value of 4.0, 0.05 g of ethylenediaminetetraacetic acid, 1 g of NaCl, and 0.1 mol of surfactant Brij 35 g, 0.1 g of isopropanol as a stabilizer, 0.1 g of chlorinated p-nitrobenzene α-L-fucopyranoside, and the balance is water.

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Abstract

The invention discloses a liquid single reagent for detecting alpha-L-fucosidase and a preparation method thereof. The liquid single reagent contains a buffer solution, ethylenediamine tetraacetic acid, NaCl, a surface active agent, a stabilizer and a P-nitrobenzene chloride alpha-L-fucus pyranoside. The stabilizer contained in the liquid single reagent disclosed by the invention can not only effectively enhance the stability of the liquid single reagent, but also effectively eliminate the interference of sample heparin by being cooperated with the surface active agent, therefore detection accuracy and sensitivity are enhanced. A stability test result indicates that after the liquid single reagent for detecting the alpha-L-fucosidase is respectively preserved for 3 days at 42 DEG C, preserved for 14 days at 37 DEG C and preserved for 18 months at 2-8 DEG C, various properties are kept stable so as to indicate that the liquid single reagent disclosed by the invention has good stability and can resist the interference of at least 100 KU / L heparin contained in a sample.

Description

technical field [0001] The invention relates to a reagent for detecting enzyme content in a human body, in particular to a liquid single reagent for detecting α-L-fucosidase and a preparation method thereof, belonging to the detection field of α-L-fucosidase. Background technique [0002] α-L-Fucosidase (α-L-Fucosidase, AFU) is a lysosomal acid hydrolase, classified as α-L-fucoside fucosidase (EC3.2.1.51). It mainly exists in various tissues, cells and body fluids in the human body, such as lysosomes of liver, pancreas, brain, lung, kidney fibroblasts, etc., with the highest content in liver and kidney. This enzyme is contained in serum, urine, saliva, tears and pituitary fluid. The basic physiological function is to catalyze the catabolism of fucosyl-containing oligosaccharides, glycopeptides, glycoproteins and glycosides. [0003] People with fucosidosis have deficiencies of the enzyme in the brain, liver, kidneys, lungs, and urine, resulting in abnormal deposits in thes...

Claims

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Application Information

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IPC IPC(8): G01N21/31
Inventor 不公告发明人
Owner BEIJING LEADMAN BIOCHEM
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