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Method for preparing angiogenesis inhibition factor

An angiogenesis inhibitory factor technology, applied in growth factors/inducing factors, animal/human proteins, medical raw materials derived from fish food, etc., can solve the problem of many differences in active protein components and structures, and the angiogenesis of red ray cartilage that has not yet formed Inhibitor factor preparation methods and other issues to achieve the effect of inhibiting angiogenesis

Inactive Publication Date: 2008-06-18
ZHEJIANG OCEAN UNIV
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Problems solved by technology

However, due to the many differences in active protein components and structures in red ray and shark cartilage, there is currently no preparation method for red ray cartilage angiogenesis inhibitors.

Method used

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  • Method for preparing angiogenesis inhibition factor
  • Method for preparing angiogenesis inhibition factor
  • Method for preparing angiogenesis inhibition factor

Examples

Experimental program
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Effect test

Embodiment Construction

[0022] The preparation method of the angiogenesis inhibitory factor provided in this embodiment, its process is

[0023] 1. Crush the red stingray cartilage.

[0024] 2. Mix 1000g of red ray cartilage fragments with 1000mL extract, which contains 8mol MES, 20g EDTA160mol guanidine hydrochloride, then adjust the pH value of the mixture to 6, shake at 20°C for 72h, and then Centrifuge at 4°C and 10,000r / min for 25min, discard the residue, and continue centrifuging at 4°C and 10,000r / min for 25min with the obtained supernatant, then discard the residue to obtain the supernatant.

[0025] 3. The supernatant obtained in step 2 was obtained by ultrafiltration to obtain the crude extract of red ray cartilage angiogenesis inhibitory factor with a molecular weight of 3-200KD, and then used 25%, 35%, 40%, 50%, 60%, and 70% , 80% (weight / volume percentage) of acetone carries out fractional precipitation to the obtained red ray cartilage crude extract, and after the precipitates are mixe...

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Abstract

The present invention provides a preparation method of angiogenesis inhibitory factor. Dasyatis cartilage is firstly crashed, then extracted, roughly separated and purified, finally desalinated, frozen and dried, thus obtaining the dasyatis cartilage angiogenesis inhibitory factor, shortened as DCAIF-I; wherein, the purification adopts Hitrap DEAEFF ion-exchange column chromatography, Superdex 75 10 / 300 GL ion-exchange column chromatography and Shim-packVP-ODS HPLC ion-exchange column chromatography. The preparation method of angiogenesis inhibitory factor provided by the present invention provides a method of extracting the angiogenesis inhibitory factor from the dasyatis cartilage. Through experiment, the DCAIF-I obtained in the method can effectively inhibit the angiogenesis and has a certain concentration dependence. The preparation method is a method in which the angiogenesis inhibitory factor can be effectively and optimally distilled from the dasyatis cartilage.

Description

technical field [0001] The invention relates to a preparation method of angiogenesis inhibitory factor, in particular to a method for preparing a protein capable of inhibiting angiogenesis from red ray cartilage. Background technique [0002] The growth, invasion, and metastasis of primary tumors all depend on tumor angiogenesis, and it is possible to effectively control tumor growth and metastasis by inhibiting angiogenesis and using tumor blood vessels as a new target for cancer treatment. Many researchers have found that there are active factors that inhibit tumor growth with different molecular weights, all of which have certain angiogenesis inhibitory activities, most of which come from shark cartilage. For example, Chinese invention patent 98111074.6 "Shark cartilage angiogenesis inhibitory factor and its separation and purification method" proposes a shark cartilage angiogenesis inhibitory factor, which is "prepared from shark cartilage through separation and purifica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/475A61K35/60
Inventor 罗红宇余新威
Owner ZHEJIANG OCEAN UNIV
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