Method for separating purified immuno regulation active polypeptide from placenta of cattle

An immunoregulatory activity, separation and purification technology, applied in the field of biomedical extraction, can solve the problem of few reports on the separation and purification of active ingredients

Inactive Publication Date: 2005-10-26
浙江三鹿实业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on placental immunomodulatory factors is mainly focused on the function and application, but there are few reports on the separation and purification of its main active components

Method used

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  • Method for separating purified immuno regulation active polypeptide from placenta of cattle
  • Method for separating purified immuno regulation active polypeptide from placenta of cattle
  • Method for separating purified immuno regulation active polypeptide from placenta of cattle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] A. Placenta treatment:

[0043] Take fresh bovine placenta, wash and cut into pieces, add twice (W / V) phosphate buffer solution with pH 6-8-7.4, homogenate, centrifuge at 12000r / m, precipitate, and take the supernatant through 10,000 molecular weight cut-off ultrafiltration , Desalting with aromatic polyamide nanofiltration membrane, and freeze-drying to obtain sample freeze-dried powder containing immunomodulatory active polypeptide;

[0044] B. Anion exchange chromatography:

[0045] Take 30mg freeze-dried powder, dissolve in 5ml pH 7.4 20mmol / L Na 2 HPO 4 -NaH 2 PO 4 In the buffer solution, load the sample on the DEAE Sepharose CL-6B anion exchange column (Pharmacia company) of 2.6 × 35cm, the flow rate is 1mL / min, and the Na2HPO4-NaH2PO4 buffer solution of gradient elution eluent A liquid is 20mmol / L, Liquid B is a solution in which 1.0mol / L sodium chloride is added to liquid A. Gradient mode is:

[0046] 0-600min Liquid A

[0047] 600-1000min B solution 0-1...

Embodiment 2

[0060] Ion exchange chromatography and gel exclusion chromatography choose pH 6.8 Na 2 HPO 4 -NaH 2 PO 4 Buffer solution; Other operating techniques are the same as in Example 1.

Embodiment 3

[0062] Ion exchange chromatography and gel exclusion chromatography choose pH 7.2 Na 2 HPO 4 -NaH 2 PO 4 Buffer solution; Other operating techniques are the same as in Example 1.

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Abstract

The present invention provides a new placenta resource-bovine placanta, and adopts the processes of cleaning cutting into block, homogenization, extraction, centrifugal precipitation separation, ultrafiltration of supernatant fluid, nano-fitlering to remove salt and freeze-drying so as to obtain the basic product containing active immuno regulatory activity polypeptide, then making said basic product undergo the processes of separation and purification by means of anion-exchange chromatography, gel exclusion chromatography and inversed phase high-performance liquid chromatography and freeze-drying so as to obtain the purified product.

Description

technical field [0001] The invention relates to a method for separating and purifying immunoregulatory active polypeptides from bovine placenta, and belongs to the technical field of biomedicine extraction. Background technique [0002] The study of placental immune regulatory factors is the first in my country, and it is an active ingredient extracted from the placenta of healthy puerpera. In 1985, Liu Yuexin first reported a small molecular active substance extracted from the placenta of healthy puerperas by using the "homogenization-dialysis" method, and called this factor placental factor; in 1994, Huang Chuhua et al. washed and cut the fresh placenta Cut, add 2 times normal saline tissue homogenate, centrifuge, supernatant is ultrafiltered with ultrafiltration membrane, and the filtrate is degermed and subpackaged to obtain placenta injection; in 1996, Zhang Xuerong et al. reported that fresh placenta was cut off fascia , rinsed with sterile water, cut into pieces and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/50C07K2/00C07K14/47
CPCC07K14/4715A61K38/00A61K35/50A61P37/02
Inventor 王玉良生庆海夏文水房新平魏鹏翟红梅
Owner 浙江三鹿实业有限公司
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