Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Preparation method of natural Sipunculus fibrinolysin and application thereof

A kind of technology of star worm and plasmin, which can be applied in the directions of biochemical equipment and methods, medical preparations containing active ingredients, enzymes, etc., and can solve problems such as unsatisfactory

Pending Publication Date: 2019-02-01
许瑞安
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Second-generation thrombolytics have improved, but are not ideal
The results of this study have not been reported so far

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Preparation method of natural Sipunculus fibrinolysin and application thereof
  • Preparation method of natural Sipunculus fibrinolysin and application thereof
  • Preparation method of natural Sipunculus fibrinolysin and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Embodiment 1 Ammonium sulfate precipitation

[0079] Put the fresh star worms in seawater for 2-3 days to discharge the visceral sediment. According to the results of active site detection, intestinal tissue was taken, added a certain volume of 0.02mol / L Tris-HCl buffer solution (pH 7.4), homogenized, taken out at 4°C, refrigerated and centrifuged at 10000rpm for 20min, and discarded the precipitate. Measure the volume of the supernatant and mix it with saturated ammonium sulfate of pH 7.4 to make the concentration of ammonium sulfate in the solution reach 30%. Stir and let it stand for 12 hours. A small amount of precipitates can be seen. 4°C, 10,000rpm, refrigerated centrifugation for 20min, the precipitate was mainly cell debris and other insoluble substances, discarded, collected the supernatant, continued to add ammonium sulfate to make the final concentration reach 90%, it can be seen that a large amount of protein precipitated, 4°C, 10,000rpm, refrigerated centri...

Embodiment 2

[0080] Example 2 Desalting of Sephadex G-25

[0081] The collected precipitate was dissolved in a small amount of 0.02mol / L Tris-HCl buffer solution (pH 7.4), and filtered through a 0.22 micron membrane filter. Sephadex G-25 desalting column pretreatment: 5 column volumes of deionized water 5ml / min to wash the column; 5 column volumes of 0.02mol / LTris-HCl buffer (pH 7.4) equilibration; sample loading: the loading volume should not be greater than the column volume 20%; elution: 5ml / min0.02mol / L Tris-HCl buffer (pH 7.4) for 1.5 column volumes, 5ml / tube set; column storage: 5ml / min0.02mol / L Tris-HCl buffer (pH 7.4) two column volumes wash 5ml / min deionized water two column volumes wash, 5ml / min 20% two column volumes wash.

Embodiment 3

[0082] Example 3 DEAE Sepharose Fast Flow Chromatography

[0083] DEAE column pretreatment: wash 10 column volumes with 2ml / min deionized water, and wash 5 column volumes with 2ml / min 0.02mol / L Tris-HCl buffer (pH 8.0). Sample loading: the sample needs to pass through a 0.22 micron filter membrane, and the sample loading flow rate is 1ml / min; equilibrium: 0.02mol / L Tris-HCl buffer (pH 8.0) 2ml / min washes five column volumes; elution: fractional elution : Use 2ml / min to elute at 0.15M, 0.2M, 0.25M, 0.3M, 0.35M, 0.4M, 0.45M, 0.5M NaCl respectively, collect 1ml / tube, and get five main peaks. The activity of collected proteins was detected by fiber plate method.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Molecular weightaaaaaaaaaa
Molecular weightaaaaaaaaaa
Molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention discloses a preparation method of natural Sipunculus fibrinolysin and application thereof. The natural Sipunculus fibrinolysin has a partial amino acid sequence of SEQ ID NO: 1. The natural Sipunculus fibrinolysin has a very good antithrombotic curative effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of natural Acanthus fibrinolytic enzyme and its pharmaceutical application. Background technique [0002] Thrombosis is a very important protective mechanism in the normal physiological response of an individual. When the body suffers trauma, it can effectively stop bleeding and achieve a protective effect. There are 3 main factors affecting hemostasis and thrombus formation: intravascular blood flow velocity, platelets and coagulation factors. Compared with physiological hemostasis, pathological thrombosis occurs in the lumen of blood vessels. If it involves arteries, it will lead to organ and tissue ischemia and / or necrosis; if it occurs in veins, it will cause blood return obstruction. The thrombus formed in one part can also travel to blood vessels in other parts along the direction of blood flow. This process is called embolism. The inner wall ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N9/64A61K38/48A61P7/02
CPCA61K38/00A61P7/02C12N9/6416C12Y304/24072
Inventor 许瑞安马国兴陈磊崔秀灵
Owner 许瑞安
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com