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Method for purifying ulinastatin based on cation exchange resin

A technology of cation exchange and ulinastatin, which is applied in the field of purification of ulinastatin based on cation exchange resin, can solve the problems of cation exchange resin purification, improve recovery efficiency and purity, shorten the process flow, and shorten the purification process Effect

Inactive Publication Date: 2016-07-13
GUANGDONG TECHPOOL BIO-PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] But up to now, there is no related report that cation exchange resin is used for the purification of ulinastatin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1) Get 1t of clear urine, adjust the pH to 6, add 8kg of silica gel, stir, filter with diatomaceous earth as a medium, get the filtrate, adjust the pH of the filtrate to 3, add 8kg of chitosan (chitosan) in the filtrate The molecular weight is 50,000, the degree of deacetylation is 93%) and 0.4kg potassium alginate (molecular weight is 50,000), stirred for 1h to obtain the mixed solution 1, and the mixed solution 1 was prepared at a pressure of 0.1MPa, a temperature of 30°C, and The molecular weight is 7×10 4 Under the condition of ultrafiltration, the retentate 1 is obtained;

[0034] 2) Adjust the pH of the retentate 1 to 7.5, filter with diatomaceous earth to obtain the filtrate 1, adjust the pH of the filtrate 1 to 4, and then flow through the cation exchange resin (AmberliteIR-120Na:AmberliteIRC54 mass ratio of 5:1 mixing), collecting the effluent to obtain effluent A;

[0035] 3) Dilute the effluent A at a pressure of 0.2 MPa, pH=7.2, and a molecular weight cut-...

Embodiment 2

[0042] 1) Get 1t of clarified urine, adjust the pH to 6.5, add 12kg of silica gel, stir, and filter with diatomaceous earth as a medium to obtain the filtrate, adjust the pH of the filtrate to 4.5, add 12kg of chitosan (chitosan) in the filtrate The molecular weight is 30,000, the degree of deacetylation is 99%) and 0.24kg sodium alginate (molecular weight is 50,000), stirred for 1.5h to obtain the mixed solution 1, and the mixed solution 1 was prepared at a pressure of 0.7MPa and a temperature of 4°C. The molecular weight cut off is 5×10 4 Under the condition of ultrafiltration, the retentate 1 is obtained;

[0043] 2) Adjust the pH of the retentate 1 to 8, filter to obtain the filtrate 1, adjust the pH of the filtrate 1 to 5, and then flow through the cation exchange resin AmberliteIR120Na at a speed of 100mL / min, collect the effluent, and reconstitute the above effluent at 80mL / min The speed of min flows through the above-mentioned cation exchange resin AmberliteIR120Na, c...

Embodiment 3

[0051] 1) Get 1t of clarified urine, adjust the pH to 6.5, add 12kg of silica gel, stir, and filter with diatomaceous earth as a medium to obtain the filtrate, adjust the pH of the filtrate to 5, add 10kg of chitosan (chitosan) in the filtrate The molecular weight is 30,000, the degree of deacetylation is 95%) and 0.4kg potassium alginate (molecular weight is 50,000), stirring for 2 to obtain the mixed solution 1, the mixed solution 1 is 0.1MPa at a pressure, the temperature is 25 ℃, and the cut-off The molecular weight is 6×10 4 Under the condition of ultrafiltration, the retentate 1 is obtained;

[0052] 2) Adjust the pH of the retentate 1 to 7, filter to obtain the filtrate 1, adjust the pH of the filtrate 1 to 6, and then flow through the cation exchange resin at a speed of 120mL / min (AmberliteIR-120Na:Amberlite200 mass ratio is 5:1 mixed) , collect the effluent, flow the above effluent through the above-mentioned cation exchange resin again at a speed of 120mL / min, repea...

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Abstract

The invention belongs to the technical field of medicines, and in particular relates to a method for purifying ulinastatin based on cation exchange resin. The method comprises the following steps: mixing chitosan and alginate with primarily filtered urine, performing ultrafiltration, and purifying the trapped fluid obtained through ultrafiltration through cation exchange resin and ultrafiltration in sequence. By adoption of the method for purifying ulinastatin based on the cation exchange resin, the purification process is shortened, the yield and the purity of the ulinastatin are high, the yield of the ulinastatin is 75% or above, and the specific activity of the ulinastatin is up to 4750 IU / mg protein.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a method for purifying ulinastatin based on a cation exchange resin. Background technique [0002] Ulinastatin is an acidic glycoprotein isolated and purified from male urine. Since the two active functional regions in the structure have a broad spectrum of enzyme inhibition, it can simultaneously inhibit trypsin and phospholipase A 2 , hyaluronidase, elastase and other hydrolytic enzyme activities; moreover, the low molecular weight components formed by the decomposition of ulinastatin also have a strong inhibitory effect on hydrolytic enzymes. Ulinastatin is not only a broad-spectrum protease inhibitor, but also has the functions of inhibiting the production of myocardial inhibitory factors, improving the circulatory state during shock, stabilizing lysosomal membranes, and inhibiting the release of inflammatory mediators. The results of a large number of clinical...

Claims

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Application Information

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IPC IPC(8): C07K14/81C07K1/36C07K1/34C07K1/18
CPCC07K14/8114
Inventor 王旭郑少亮田友军肖益热
Owner GUANGDONG TECHPOOL BIO-PHARMA CO LTD
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