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A kind of production method of acarina

A production method and technology of cypressase are applied in the production field of cypressase, which can solve the problems of long production cycle, complex production process, long production process cycle, etc., and achieve good production process reproducibility and short production cycle. Effect

Active Publication Date: 2016-08-31
FUJIAN HUITIAN BIOLOGICAL PHARMA
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Problems solved by technology

The purity of the Aggressin prepared by this production process needs to be further improved, and the production process is complicated, the production cycle is long, and it is not easy to promote industrialization
[0004] In the prior art, there is also a target component that is initially obtained by gel filtration chromatography and then ion exchange chromatography. This component is detected as a protein component by gel electrophoresis, which is actually a molecular weight and isoelectric component. Mixtures of proteins or peptides with similar points need to be further purified, that is, there is a problem that the purity of the obtained acutinase is not high enough
In addition, the existing production technology of Agkistrodon enzyme generally has the problem of long production process cycle

Method used

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  • A kind of production method of acarina
  • A kind of production method of acarina
  • A kind of production method of acarina

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Embodiment 1

[0043]Step 1: At a temperature of 4-10°C, dissolve the lyophilized powder of Achilles venom in an excess of 0.04-0.06 mol / L Tris-HCl buffer solution with a pH value of 8-9, let it stand for more than 60 minutes, 0- Centrifuge at 10°C to obtain the supernatant, put the supernatant into a dialysis bag (molecular weight cut-off 7000 Daltons), and dialyze with 0.01mol / L Tris-HCl buffer solution with a pH value of 8-9 to remove salt and Impurities of small biological molecules; filter the dialyzed Agritus venom solution with a 0.22 μm microporous membrane.

[0044] Step 2: The solution obtained in step 1 was eluted and separated with a DEAE‐Sepharose Fast Flow anion exchange column to remove hematological substances. figure 1 It is the chromatogram of DEAE‐Sepharose Fast Flow column chromatography, the abscissa is time (min), and the ordinate is response value (mAU). Such as figure 1 As shown, 11 protein peaks were obtained, and the peak of VIII and the first half peak of IX with...

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Abstract

The invention relates to a production method of acutase. The production method comprises the following steps: (1) dissolving long-noded pit viper venom freeze-dried powder in excess 0.05mol / L Tris-HCL buffer solution with the pH value of 8.5; (2) putting the long-noded pit viper venom solution on a DEAT-Sepharose Fast Flow anion-exchange column to perform elution separation; (3) putting the solution obtained in the step (2) on a SOURCE 30Q gel column to perform chromatography, and then performing separation by virtue of phenyl sepharose high performance column chromatography; and (4) putting the solution obtained in the step (3) on a Hiload 26 / 60Superdex75PG column, and collecting a second protein peak with thrombin-like enzyme activity, thereby obtaining an acutase solution. The acutase solution prepared by the production method disclosed by the invention is a single component and has the advantage of high purity.

Description

technical field [0001] The invention relates to the field of biopharmaceuticals, in particular to a method for producing an acarina enzyme. Background technique [0002] Achilles enzyme is a thrombin-like enzyme isolated and purified from Agkistrodon venom, and it is a mixture of serine-type proteolytic enzymes. Aggressin has thrombin-like enzyme activity, its molecular weight is 24,000-30,000 Daltons, and its isoelectric point is 4.5-5.0. Achilles enzyme preparation--Acutesin injection can be used to treat acute cerebral infarction and various thrombotic diseases, and it is administered clinically by intravenous infusion. [0003] There are existing documents about the production process of acarina, and the Chinese patent application publication number is CN1229136A. The patent document discloses acarina and its production process, dissolves the upper Sephadex G‐75 column with a known buffer solution, and collects blood coagulation products. Peaks A and B of enzyme-like e...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/64
CPCC12N9/6418
Inventor 万兴平蒋宗解陈碧强李玉洁宁千年余成恢
Owner FUJIAN HUITIAN BIOLOGICAL PHARMA
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