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Method of extracting and preparing deer DGF

A technology of epidermal growth factor and deer antler, which is applied in the field of medical or edible products of deer antler extract, and can solve problems such as restriction of dosage, waste of resources, protein denaturation such as EGF, etc.

Active Publication Date: 2007-10-17
海宁市紫金港生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because the wine contains a high concentration of alcohol as an organic solvent, it will denature proteins such as EGF, and it is not conducive to dissolving proteins such as EGF from large-grain velvet antler, which will lead to low utilization efficiency of EGF in velvet antler, resulting in waste of resources.
In addition, medicinal wine with high alcohol content also limits the amount of each dose

Method used

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  • Method of extracting and preparing deer DGF
  • Method of extracting and preparing deer DGF

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1, Pretreatment of velvet antler

[0028] Take 0.5 kg of fresh velvet antler, use a cryostat to cut the velvet antler into slices with a thickness of 1 to 2 mm, and then add an appropriate amount (200 mL) of pre-cooled deionized water to crush with a homogenizer at 4°C, and proceed to homogenization. It ends when there are no visible tissue blocks in the slurry. Then, 200 mL of pre-cooled deionized water was added to the homogenate, and the mixture was mixed and ultrasonically broken under ice bath conditions. The resulting crushed product was centrifuged at 5000 revolutions per minute (rpm) for 20 minutes, and the supernatant was taken. Add 200 mL of pre-cooled deionized water to the centrifugal pellet, resuspend the pellet and centrifuge again at 5000 rpm for 20 minutes, and take the supernatant. Then, repeat the above resuspension and centrifugation process once, and take the supernatant. Combine the supernatants obtained from each centrifugal separation, put the...

Embodiment 2

[0029] Example 2. Chromatographic purification of velvet antler epidermal growth factor and preparation of lyophilized agent

[0030] Take 820 mL of the deer antler extract obtained in Example 1 and apply it to Q Sepharose TM Fast Flow column (column parameter 5.5cm×20cm) (purchased from Amersham), and then eluted. During elution, the impurity protein peak is eluted with 300mL 0.02mol / L Tris-HCl buffer (pH8.0) at a flow rate of 2mL / min, and then 450mL 0.02mol / L Tris containing 0.5mol / L NaCl -HCl buffer (pH 8.0) for elution, and use an ultraviolet detector to detect at a wavelength of 280 nm (the elution profile is shown in Figure 1). The product obtained by dialysis of each eluted fraction was detected by the detection method described in Example 3, and it was found that the peak referred to by the "target protein peak" in FIG. 1 was velvet antler epidermal growth factor. Therefore, collect the target protein peak, put it into a dialysis bag with a molecular weight cut-off of 3500...

Embodiment 3

[0031] Example 3. Detection of epidermal growth factor

[0032] According to the "Molecular Cloning Test Guide" (Science Press, 2002) and the biological activity detection method of Xiong Sheng et al. (Chinese Journal of Health Inspection, 2006, 16(10): 1153-1155, 1174), velvet epidermal growth factor was detected. The velvet antler epidermal growth factor solution obtained in Example 2 was subjected to SDS-PAGE (15% polyacrylamide separation gel, 4% layered gel, 10mA constant current electrophoresis), and stained with Coomassie brilliant blue after the electrophoresis. As shown in Figure 2, there is a single band of epidermal growth factor at a molecular weight of about 7kD, and its purity is 97.2% by image analysis and area normalization method. The proliferation of epidermal cells was used to detect the biological activity of EGF. The blank group added with buffer was used as a control. In the experimental group added with the velvet antler epidermal growth factor solution obta...

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Abstract

The present invention relates to method for extracting epidermal growth factor from deer antler which comprise : carrying out anion exchange chromatography to deer antler leachate, obtaining high purity deer antler epidermal growth factor without any other chromatography step. In addition, The present invention also relates to deer antler extract enriched with EGF prepared from the method and its uses in drug, food and cosmetics.

Description

Technical field [0001] The invention belongs to the field of medical or edible products of deer antler extract. More specifically, the present invention relates to a method for extracting epidermal growth factor (EGF) from deer antler and a deer antler extract rich in epidermal growth factor prepared by the method. In addition, the present invention also relates to the application of the deer antler extract in medicine, food and cosmetics. Background technique [0002] Deer antler (deer antlers that have not yet been ossified) and antlers are traditional Chinese medicinal materials, which have the functions of strengthening muscles and bones, invigorating the kidney and strengthening yang, and dredging blood vessels. Since ancient times, they have been widely used to formulate various nourishing drugs, foods, and health products. And cosmetics. Through modern medicinal chemistry research and analysis, velvet antler contains many kinds of physiologically act...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/60C07K1/16A61K38/25A61K9/19A61P13/12A61P21/00A61K8/64A23L1/305A23K1/165A23L33/17
Inventor 朱成钢王利忠
Owner 海宁市紫金港生物科技有限公司
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