CN1721440A - Process for preparing thymosin - Google Patents

Abstract

The present invention belongs to the field of biomedicine preparing technology. The preparation process of thymulin includes the following steps: crushing fresh thymus tissue at 20-23 deg.c inside high speed shearing machine to obtain homogeneous slurry; adding dilute to regulate pH value to 4.5-4.8; setting the homogeneous slurry at 20-23 deg.c for 3-4 hr; heating in 60-65 deg.c for 30 min and cooling naturally to room temperature; filtering; purifying the filtrate with hollow fiber column with intercepted molecular weight 10 kDa and disinfection to obtain the product. The present invention has the advantages of simple process, high yield, low production cost, easy use in large scale production, etc.

Description

The preparation method of Zadaxin

Technical field

The present invention relates to a kind of technology of preparing of biologics, be meant a kind of preparation method of Zadaxin especially.

Background technology

Zadaxin has immunoregulation effect, it is the small-molecule substance that multiple peptide class is arranged that adopts the biochemical method extraction from the thymic tissue of animal, traditional Zadaxin preparation technology and improved technology method generally include following steps: (1) with mincer or colloidal mill with fresh thymus gland homogenate, and carry out multigelation 2～3 times, with smudge cells; Add PH2.0～4.0 buffered soln then, and using ultrasonication below 10 ℃.(2) homogenate is heated to 70 ℃～85 ℃, and kept 10 minutes, centrifugal removal albumen precipitation.(3) centrifugate is carried out ultrafiltration with the ultrafiltration post of 6000～10000 molecular weight that dam.(4) ultrafiltrated is filtered or nanofiltration concentrates and to obtain product.There is the deficiency that facility investment is big, the process time is long, extracted amount is low, production cost is high in processes such as above-mentioned processing method need pass through multigelation, centrifugation, ultrafiltration, concentrate.

Summary of the invention

The objective of the invention is provides the preparation method of a kind of Zadaxin that a kind of facility investment is little, the process time is short, extracted amount is high, production cost is low for overcoming above-mentioned technology deficiency.

The object of the present invention is achieved like this: a kind of preparation method of Zadaxin may further comprise the steps:

(1) fresh thymic tissue is obtained homogenate with the high speed shear crusher machine down at 20 ℃～23 ℃;

(2) in homogenate, add dilute hydrochloric acid and regulate pH value between 4.5～4.8;

(3) homogenate that will regulate behind the pH value was placed 3～4 hours under 20 ℃～23 ℃ conditions;

(4) aforesaid liquid was heated 30 minutes in 60 ℃～65 ℃ water-baths, be chilled to room temperature naturally;

(5) mixed solution is filtered;

(6) with filtrate after with the tubular fibre column purification, degerming of the molecular weight 10000 that dams product.

The present invention compared with prior art has following characteristics:

(1) with high-speed shearing machine homogenate thymus gland, can make it broken fully, avoid ultrasonic wave active and influence to Zadaxin.

(2) pH value of mixing well slurries with hydrochloric acid is 4.5～4.8, can make the foreign protein precipitation, and inhibitory enzyme improves the activity of Zadaxin to the destruction of Zadaxin.

(3) filter with filter cloth or canvas, save facility investment, reduce operation and consume.

(4) do not use concentration technique, the content of Zadaxin is not less than 14mg/ml in the extracting solution that obtains.Reduce the equipment input, shortened the process time.

Embodiment

Get 1kg fresh calf thymus gland, add 1000 milliliters of injection liquid waters at 20 ℃～23 ℃ with high-speed shearing machine homogenate 15 minutes, until becoming milky white liquid.Add dilute hydrochloric acid and transfer pH value to 4.5～4.8, place to be placed in 3 hours in 60 ℃～65 ℃ water-baths and heated 30 minutes, filter with canvas, filtrate is through the tubular fibre column purification of 10000 molecular weight that dam, degerming promptly gets thymus peptide solution 700ml, and surveying Zadaxin content is 15mg/ml.

Claims (1)

1. the preparation method of a Zadaxin, its feature comprises the steps:

(1) fresh thymic tissue is obtained homogenate with the high speed shear crusher machine down at 20 ℃～23 ℃;

(2) in homogenate, add dilute hydrochloric acid and regulate pH value between 4.5～4.8;

(3) homogenate that will regulate behind the pH value was placed 3～4 hours under 20 ℃～23 ℃ conditions;

(4) aforesaid liquid was heated 30 minutes in 60 ℃～65 ℃ water-baths, be chilled to room temperature naturally;

(5) mixed solution is filtered;

(6) with filtrate after with the tubular fibre column purification, degerming of the molecular weight 10000 that dams product.