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A method for simultaneously preparing crude human urinary kininogenase and crude human urinary trypsin inhibitor

A technology of urinary kallikrein and urinary trypsin, which is applied in the directions of biochemical equipment and methods, peptidase, enzyme, etc., can solve the problems of unfavorable industrial production, low KN yield, etc., and achieves convenient purification treatment and reduced production cost. , the effect of improving the purification rate

Active Publication Date: 2017-10-27
GUANGDONG TECHPOOL BIO-PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But, the KN yield that this method obtains is lower, is unfavorable for industrialized production

Method used

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  • A method for simultaneously preparing crude human urinary kininogenase and crude human urinary trypsin inhibitor
  • A method for simultaneously preparing crude human urinary kininogenase and crude human urinary trypsin inhibitor
  • A method for simultaneously preparing crude human urinary kininogenase and crude human urinary trypsin inhibitor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] S1 Place 100 kg of strong base anion exchange resin as an adsorbent in a urine tank or urinal, and collect the strong base anion exchange resin that has absorbed urine protein;

[0033] S2 elutes the strong base anion exchange resin obtained in step S1 with a NaCl solution whose concentration is 0.72mol / L, and collects the eluent;

[0034] S3 Concentrate the eluate obtained in step S2 by ultrafiltration, adjust the pH value of the ultrafiltration concentrate to 3.9, and the conductivity to 2.3Ms / cm;

[0035] S4 load the concentrated solution obtained in step S3 onto the strong-acid cation exchange resin that has been balanced with the balance solution, wash it with 0.1mol / L acetic acid-sodium acetate buffer solution, the conductivity is 3Ms / cm, and the pH value is 3.9. The equilibrium solution of the strong acid type cation exchange resin is 0.1mol / L acetic acid-sodium acetate buffer solution, the conductance is 2.3Ms / cm, and the pH value is 3.9;

[0036] S5 was eluted...

Embodiment 2

[0039] S1 Place 100 kg of strong base anion exchange resin as an adsorbent in a urine tank or urinal, and collect the strong base anion exchange resin that has absorbed urine protein;

[0040] S2 elutes the strong base anion exchange resin obtained in step S1 with a NaCl solution whose concentration is 0.6mol / L, and collects the eluate;

[0041] S3 Concentrate the eluate obtained in step S2 by ultrafiltration, adjust the pH value of the ultrafiltration concentrate to 3.2, and conductance to 4.3Ms / cm;

[0042]S4 Load the concentrated solution obtained in step S3 onto the strong-acid cation exchange resin that has been balanced with the balance solution, and wash it with 0.3mol / L acetic acid-sodium acetate buffer solution, the conductivity is 4.1Ms / cm, and the pH value is 3.2. , the equilibrium solution of the strong acid type cation exchange resin is 0.3mol / L acetic acid-sodium acetate buffer solution, the conductance is 4.1Ms / cm, and the pH value is 3.2;

[0043] S5 was elute...

Embodiment 3

[0046] S1 Place 100 kg of strong base anion exchange resin as an adsorbent in a urine tank or urinal, and collect the strong base anion exchange resin that has absorbed urine protein;

[0047] S2 elutes the strong base anion exchange resin obtained in step S1 with a NaCl solution whose concentration is 0.8mol / L, and collects the eluate;

[0048] S3 Concentrate the eluate obtained in step S2 by ultrafiltration, adjust the pH value of the ultrafiltration concentrate to 4.2, and conductance to 1.3Ms / cm;

[0049] S4 Apply the concentrated solution obtained in step S3 to the strong-acid cation exchange resin that has been balanced with the balance solution, and wash it with 0.02mol / L acetic acid-sodium acetate buffer solution, the conductivity is 1.2Ms / cm, and the pH value is 4.2. , the equilibrium solution of the strong acid type cation exchange resin is 0.02mol / L acetic acid-sodium acetate buffer solution, the conductance is 1.2Ms / cm, and the pH value is 4.2;

[0050] S5 was elu...

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Abstract

The invention relates to a urine protein extraction method, in particular to a method for preparing a crude human urine kininogenase product.According to the method for preparing the crude human urine kininogenase product, anion exchange resin is adopted as an adsorbent to adsorb urine protein in urine, then the urine protein adsorbing resin is collected and eluted in a centralized mode, the eluant is used for adsorbing human urine kininogenase through cation exchange resin, and therefore human urine kininogenase and a human urine trypsin inhibitor are separated.Through the method for preparing the crude human urine kininogenase product, human urine kininogenase and the human urine trypsin inhibitor can be effectively separated, subsequent purification of human urine kininogenase and the human urine trypsin inhibitor is convenient, the purification rate is increased, co-production of two crude protein products is realized, and production cost can be greatly reduced.

Description

technical field [0001] The invention relates to a method for extracting urine protein, in particular to a method for preparing crude human urinary kininogenase. Background technique [0002] There are more than 300 kinds of proteins in human urine. At present, urokinase, human urinary trypsin inhibitor and human urinary kininogenase have been developed and extracted. These components have important therapeutic value in clinic. [0003] Urinary Kallidinogenase (KN for short) is a glycoprotein composed of 238 amino acids isolated from human urine, with an isoelectric point of about 4 and a molecular weight of about 54,000D, belonging to serine proteases. It activates the conversion of human plasma kininogen to kinin. At present, it has been developed as a drug for the treatment of acute cerebral infarction. [0004] Urinary trypsin inhibitor (UTI for short) is a glycoprotein composed of 143 amino acids isolated and purified from human urine, with an isoelectric point of abou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/64
CPCC12N9/6424C12Y304/21106
Inventor 王旭章华何建国曾永峰
Owner GUANGDONG TECHPOOL BIO-PHARMA CO LTD
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