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Method for extracting and purifying calf serum protein-removing extract

A calf serum, deproteinized technology, applied in the directions of drug combination, medical raw materials derived from mammals, metabolic diseases, etc., can solve problems such as low activity, potential safety hazards, and inability to guarantee the biological safety of products, and achieve broad application prospects, Guaranteed safety, improved purity and biological activity

Active Publication Date: 2008-03-12
JINZHOU AHON PHARM CO LTD
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Problems solved by technology

However, the product extracted by the above method has low purity and low activity, and no virus inactivation is carried out, so the biological safety of the product cannot be guaranteed (such as cattle may carry multiple viruses such as mad cow disease and foot-and-mouth disease), which brings the safety of clinical application. Hidden danger

Method used

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  • Method for extracting and purifying calf serum protein-removing extract

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Embodiment 1

[0012] Embodiment 1, extract calf serum deproteinized extract

[0013] 1. Extraction and purification method

[0014] (1) Take venous blood from young cattle and separate the serum:

[0015] Under sterile conditions, collect venous blood from calves less than 6 months old, put it in a refrigerator at 4°C overnight, let it naturally separate part of the serum, and centrifuge the rest at 4000rpm, 4°C for 15 minutes under sterile conditions to separate the serum spare.

[0016] (2) Ethanol removes protein:

[0017] Add 96% ethanol to the serum according to the volume ratio of ethanol and serum of 2:1, stir constantly, let it stand for 60 minutes to denature and precipitate the protein, centrifuge (4000 rpm, 10 minutes) and leave the supernatant to discard the precipitate.

[0018] (3) In addition to ethanol:

[0019] Use a vacuum rotary evaporator to remove ethanol under reduced pressure at 37°C and 1 atmosphere.

[0020] (4) Coarse filtration:

[0021] Roughly filter with ...

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Abstract

The invention discloses a method for extracting and purifying calf serum protein-removing extract, which comprises extracting calf venous blood, separating plasma, removing protein with ethanol, removing ethanol through decompression, charging composite proteolytic enzyme, subjecting supernatant fluid to ultrafiltration, desalinizing ultrafiltrate with gluglucosan G-15 gel chromatography, collecting elution portion with specific absorption at 280nm, then separating with DEAE-Sephadex-50 gel chromatography, collecting elution portion with specific absorption at a second 280nm, then carrying out virus deactivation by means of microporous membrane filtration and ultraviolet irradiation.

Description

technical field [0001] The invention relates to a method for extracting and purifying calf serum deproteinized extract. Background technique [0002] Calf serum deproteinized extract (protein-free calf blood extract) is a small molecule biologically active substance obtained from 1-6 month old calf serum after deproteinization, containing small molecule peptides, amino acids, keto acids and other substances. Its main pharmacological effect is to enhance the uptake and utilization of oxygen and glucose by cells, and has the physiological effect of improving cell metabolism. It is widely used clinically. [0003] At present, the commonly used extraction methods of deproteinized calf serum mainly include acid hydrolysis and enzymatic hydrolysis. Among them, acid hydrolysis will seriously damage the basic structure of the extract and seriously inactivate the product; enzymatic hydrolysis can ensure biological activity, such as China The method disclosed in the invention patent ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/16A61P3/00
Inventor 于洪儒王洪新杨菁
Owner JINZHOU AHON PHARM CO LTD
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