Purification method, extract and preparation of cobra venom neurotoxin

A technology for cobra venom and neurotoxin, which is applied to the preparation methods of peptides, nervous system diseases, chemical instruments and methods, etc., can solve the problems of difficult quality control, low purity, long production cycle, etc., and achieves improved separation and simplified extraction process. , the effect of saving time and cost

Inactive Publication Date: 2012-02-15
GUIZHOU YIBAI PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The technical problem to be solved by the present invention is to overcome the shortcomings of long production cycle, difficult quality control and low purity in the existing extraction method of cobra venom neurotoxin, and to provide a cobra venom neurotoxin with simple process, safety and quality controllable. Toxin Extraction Method

Method used

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  • Purification method, extract and preparation of cobra venom neurotoxin
  • Purification method, extract and preparation of cobra venom neurotoxin
  • Purification method, extract and preparation of cobra venom neurotoxin

Examples

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Embodiment 1

[0094] Embodiment 1: the preparation of cobra venom neurotoxin

[0095] 1) Dissolve 100g of cobra venom powder in pH7.9, 0.01M phosphate buffer to 280ml, dissolve under stirring, centrifuge at 3000r / min for 10min, take the supernatant, pass the supernatant through 0.22μm membrane;

[0096] 2) The obtained supernatant is subjected to ultrafiltration and exchanged for a separation buffer, using an ultrafiltration membrane with a molecular weight cut-off of 10KDa;

[0097] 3) The obtained snake venom is loaded onto the CM-cellulose chromatography column with pH7.9, 0.01M phosphate buffer balance, and is eluted with phosphate buffer, and the elution flow rate is 40ml / h. The eluate was collected by an automatic fraction collector and detected at 280 nm by a UV spectrophotometer.

[0098] 4) When the absorbance of the eluate drops below 0.2, elute with a phosphate buffer containing 0.04mol / L Nacl, and when the absorbance of the eluate drops below 0.2, elute with a phosphate buffer...

Embodiment 2

[0100] Embodiment 2: the preparation of cobra venom neurotoxin extract

[0101] 1) Dissolve 100g of cobra venom powder in pH 8.0, 0.01M phosphate buffer to 280ml, dissolve under stirring, centrifuge at 3000r / min for 10min, take the supernatant, pass the supernatant through 0.22μm membrane;

[0102] 2) The obtained supernatant is subjected to ultrafiltration and exchanged for a separation buffer, using an ultrafiltration membrane with a molecular weight cut-off of 10KDa;

[0103] 3) The obtained snake venom is loaded onto the CM-cellulose chromatography column with pH8.0, 0.01M phosphate buffer balance, and is eluted with phosphate buffer, and the elution flow rate is 50ml / h. The eluate was collected by an automatic fraction collector and detected at 280 nm by a UV spectrophotometer.

[0104] 4) When the absorbance of the eluent drops below 0.2, elute with a phosphate buffer containing 0.05 mol / L Nacl. After the absorbance of the eluate drops below 0.2, take the part with th...

Embodiment 3

[0106] Embodiment 3: the preparation of cobra venom neurotoxin extract

[0107] 1) Dissolve 100g of cobra venom powder in pH7.8, 0.01M phosphate buffer to 280ml, dissolve under stirring, centrifuge at 3000r / min for 10min, take the supernatant, pass the supernatant through 0.22μm membrane;

[0108] 2) The obtained supernatant is subjected to ultrafiltration and exchanged for a separation buffer, using an ultrafiltration membrane with a molecular weight cut-off of 10KDa;

[0109] 3) The obtained snake venom is loaded onto the CM-cellulose chromatographic column that has been equilibrated with pH7.8, 0.01M phosphate buffer, and is eluted with phosphate buffer, and the elution flow rate is 30ml / h. The eluate was collected by an automatic fraction collector and detected at 280 nm by a UV spectrophotometer.

[0110] 4) When the absorbance of the eluent drops below 0.2, elute with a phosphate buffer containing 0.03mol / L Nacl, and when the absorbance of the eluate drops below 0.2, e...

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Abstract

The invention discloses a purification method, an extract and a preparation of cobra venom neurotoxin. The extract of the cobra venom neurotoxin is prepared from cobra crude venom through extraction and purification. A filter membrane filtering method is firstly adopted for removing germs, particles and macromolecular sensitizers in solution, the time and the cost are saved for the subsequent processes, and the safety of products is enhanced; an ion exchange method is adopted for eluting and separating the neurotoxin, and the separation of neurotoxin protein is improved; and finally, acetone is adopted for directly precipitating the cobra venom neurotoxin, and impurities in final products are removed. Compared with the prior art, the extract and the preparation obtained in the invention have the advantages that the product yield, the content and the purity are improved, the adverse reaction of the medicine is effectively reduced, the curative effect is improved, and the quality of products is ensured.

Description

technical field [0001] The invention belongs to the technical field of biomedicine extraction, and in particular relates to a purification method, an extract and a preparation thereof of cobra snake venom neurotoxin. Background technique [0002] Snake venom is a liquid secreted by poisonous snakes from the venom glands. The main component is toxic protein, which accounts for about 90% to 95% of the dry weight. There are more than 20 kinds of enzymes and toxins. In addition, it also contains some small molecular peptides, amino acids, carbohydrates, lipids, nucleosides, biogenic amines and metal ions. The composition of snake venom is very complex, and the toxicity, pharmacology and toxicological effects of different snake venoms have their own characteristics. As far as cobra snake venom is concerned, it contains cytotoxin (CTX), neurotoxin (neurotoxin, NT), nerve growth factor (Nerve growth factor, NGF) and various enzymes. Neurotoxin (NT) has a relatively high content ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/465C07K1/36C07K1/34C07K1/30C07K1/18A61K38/17A61K35/58A61P25/04A61P29/00A61K35/583
Inventor 窦啟玲苏凯
Owner GUIZHOU YIBAI PHARMA CO LTD
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