Method for extracting single component batroxobin from Bothrops atrox poison

A technology for batroxobin and pit viper, which is applied in the field of batroxobin, can solve the problems that the literature process is difficult to achieve separation and preparation requirements, is difficult to achieve large-scale production purposes, cannot be used in actual production, and the like, and achieves easy control and convenience. Operation, the effect of a large amount of snake venom

Inactive Publication Date: 2007-11-28
QILU PHARMA
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  • Abstract
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AI Technical Summary

Benefits of technology

This new method described by this patented technology allows for better efficiency during processing without damaging or affecting certain proteins that may have important functions like cell growth. It also provides excellent separating properties even when used on small amounts of material (large quantities).

Problems solved by technology

This patents describes different ways to prepare pure biotoxysulfonate compounds called benzoapirrone sulfones, specifically those containing both esterified amygentic hydroxyproline/serum factor antigens and labeled hepatoxic substances like tissue plasminogen activator type 1(TNAP1). These complex mixtures contain many peptides involved in their functioning pathways. However, these techniques involve complicated steps involving multiple resin column processes, leading to reduced yields due to impurities during processing. Chemistry synthesis approaches were developed recently, but they still require extensive experimentations and may result in unwanted side effects on human health.

Method used

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  • Method for extracting single component batroxobin from Bothrops atrox poison

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Experimental program
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Effect test

Embodiment 1

[0039] The concrete steps of this embodiment are as follows:

[0040] 1. Take Agkistrodon venom 3g, dissolve it in 60ml Tris-HCL buffer solution with pH8.6 containing 1.0M NaCl, soak it in a refrigerator at 4°C to fully dissolve it, then centrifuge at 8000rpm for 10min, remove the precipitate, and collect the supernatant of the snake venom ;

[0041] 2, Benzamidine Sepharose 6B (Benzamidine Sepharose 6B) affinity chromatography column (2.6cm * 30cm) equilibrates at least 5 column volumes with the Tris-HCL buffer solution containing 1.0M NaCL of pH8.6, and above-mentioned snake venom Put the supernatant on the column and control the flow rate to 0.3ml / min;

[0042] 3. After loading the sample, use the Tris-HCL buffer solution containing 1.0M NaCl at pH 8.6 to elute until the reading of the nucleic acid protein detector reaches the baseline;

[0043] 4. Connect the affinity chromatography column to Sephadex G25 desalting column (1.6cm×100cm), use pH8.6 containing 0.2M NaCL and...

Embodiment 2

[0048] The concrete steps of this embodiment are as follows:

[0049] 1. Take 5g Agkistrodon venom, dissolve it in 80ml Tris-HCL buffer solution with pH 9.0 containing 0.6M NaCl, soak it in a refrigerator at 4°C to fully dissolve it, then centrifuge at 8000rpm for 10min, remove the precipitate, and collect the supernatant of the snake venom ;

[0050] 2, Benzamidine Sepharose 6B (Benzamidine Sepharose 6B) affinity chromatography column (2.6cm * 30cm) equilibrates at least 5 column volumes with the Tris-HCL buffer solution containing 0.6M NaCL of pH9.0, and above-mentioned snake venom Put the supernatant on the column and control the flow rate to 0.3ml / min;

[0051] 3. After loading the sample, use Tris-HCL buffer solution containing 0.6M NaCl at pH 9.0 to elute until the reading of the protein detector reaches the baseline;

[0052] 4. Connect the affinity chromatography column to Sephadex G25 desalting column (1.6cm×100cm), use pH 9.0 containing 0.1M NaCL and 0.2M benzamidi...

Embodiment 3

[0057] The concrete steps of this embodiment are as follows:

[0058] 1. Take 5g Agkistrodon venom, dissolve it in 80ml Tris-HCL buffer solution with pH8.0 containing 0.6M NaCl, soak it in a refrigerator at 3°C ​​to fully dissolve it, then centrifuge at 8000rpm for 10min, remove the precipitate, and collect the supernatant of the snake venom ;

[0059] 2, Benzamidine Sepharose 4B (Benzamidine Sepharose 4B) affinity chromatography column (2.6cm * 30cm) equilibrates at least 5 column volumes with the Tris-HCL buffer solution containing 0.6M NaCL of pH8.0, and above-mentioned snake venom Put the supernatant on the column and control the flow rate to 0.3ml / min;

[0060] 3. After loading the sample, elute with pH 8.0 Tris-HCL buffer containing 0.6M NaCl until the reading of the protein detector reaches the baseline;

[0061] 4. Connect the affinity chromatography column to Sephadex G25 desalting column (1.6cm×100cm), use pH 8.0 containing 0.3M NaCL and 0.15M benzamidine hydrochlo...

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Abstract

It' s a method for extracting Batroxobin for defibrination and thrombolysis from venom of snakes, which relates to medicine technologies. The method comprises the steps of soaking, dissolving and centrifuging the venom of the snake; chromatography of the supernatant by an affinity column; chromatography of the eluent that has been desalinated by flowing it through polydextran gel G25; entrapment and concentration of the eluent that containing effective compositions and chromatography of the concentrated sample by a molecular sieve so as to obtain the Batroxobin with sole composition. The method features convenient operation, short production cycle, high purity and is suitable for production in large scale.

Description

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Claims

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Application Information

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Owner QILU PHARMA
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