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Method for removing endotoxin in human urokinase raw material refining process

A technology for urokinase and endotoxin, which is applied in the directions of biochemical equipment and methods, ion exchange column/bed methods, enzymes, etc., can solve the problems of reducing production costs, complex methods, increasing costs, etc. Simple, material-saving effect

Active Publication Date: 2020-10-16
JIANGSU YOULIKA BIOTECHNOLOYG CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method of this invention is relatively complicated, and at the same time, the ultrafiltration membrane is mainly used for ultrafiltration in this method. Since most of the ultrafiltration membranes cannot be chemically treated, the service life is relatively short, and the replacement cost will be increased.
[0004] In view of the complexity, high cost, and low yield of urokinase activity in the purification method of urokinase, it is urgent to find a method for removing endotoxin from urokinase with high activity yield and excellent removal effect, so as to effectively remove the intermediate product of urokinase reduce production costs while eliminating endotoxins in

Method used

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  • Method for removing endotoxin in human urokinase raw material refining process

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Experimental program
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Effect test

Embodiment 1

[0031] A method for removing endotoxin during the refining process of human urokinase raw materials, comprising the following steps:

[0032] (1) Sample pretreatment: adjust the pH of the raw material intermediate solution of human urokinase to 6.0 and the conductance to 1ms / cm;

[0033] (2) Chromatographic column preparation and packing: add water to the TOYOPEARL ion exchange resin superQ-650M of Tosoh Co., Ltd., Japan, to obtain a suspension, pack the column, and obtain the chromatographic column; wherein, the column height of the packing is 10cm , the diameter of the chromatographic column is 1.6cm; wherein, the specific steps of packing the column are: replace the preservation solution in the chromatographic filler with water for injection, a total of three times, adjust the chromatographic filler after the last replacement: the ratio of water is 1: 0.4; Start the chromatography system, drain the air in the chromatography system and the column head screen with water for i...

Embodiment 2

[0038] A method for removing endotoxin during the refining process of human urokinase raw materials, comprising the following steps:

[0039] (1) Sample pretreatment: adjust the pH of the raw material intermediate solution of human urokinase to 9.0 and the conductance to 10ms / cm;

[0040] (2) Chromatographic column preparation and column packing: add water to the SOURCE 30Q high-speed low back pressure ion exchange medium of U.S. General Electric Company, obtain suspension, pack column, obtain chromatographic column; Wherein, the column height of packing is 30cm, and the diameter of the chromatographic column is 30cm; wherein, the specific steps of packing the column are: replace the preservation solution in the chromatographic filler with water for injection, a total of three times, adjust the chromatographic filler after the last replacement: the ratio of water is 1: 1. Start the chromatography system, drain the air in the chromatography system and the column head screen wit...

Embodiment 3

[0045] A method for removing endotoxin during the refining process of human urokinase raw materials, comprising the following steps:

[0046] (1) Sample pretreatment: adjust the pH of the raw material intermediate solution of human urokinase to 8.0 and the conductance to 5ms / cm;

[0047] (2) Chromatographic column preparation and column packing: add water to the Chromalite MQ / F weak base anion resin of British Brilliant Resin Co., Ltd. to obtain a suspension, pack the column, and obtain the chromatographic column; wherein, the column height of the packed column is 20cm, and the diameter of the chromatography column is 20cm; wherein, the specific steps of packing the column are: replace the preservation solution in the chromatography filler with water for injection, a total of three times, adjust the chromatography filler after the last replacement: the ratio of water is 1 :0.8; start the chromatography system, drain the air in the chromatography system and the column head scre...

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Abstract

The invention provides a method for removing endotoxin in a human urokinase raw material refining process, and relates to the field of bioengineering and chemical engineering, and the method specifically comprises the following steps: (1) sample pretreatment; (2) preparing and packing of a chromatographic column; and (3) disinfection and sample loading treatment. According to the method, a novel polymer anion chromatography medium is adopted, various process conditions are optimized, the treatment process is simple, endotoxin in a urokinase intermediate product can be effectively removed, andmeanwhile it is guaranteed that the urokinase activity yield reaches 75% or above. In the treatment process, working hours and materials are saved, and the production cost is reduced.

Description

technical field [0001] The invention relates to the fields of bioengineering and chemical engineering, in particular to a method for removing endotoxin during the refining process of human urokinase raw materials. Background technique [0002] Urokinase is a thrombolytic drug extracted from fresh human urine. It can activate plasminogen and convert it into active plasmin. dissolve. Therefore, it is mostly used clinically to treat thrombosis, thromboembolism and other diseases. When urokinase is used in combination with anticancer agents, because it can dissolve fibrin around cancer cells, anticancer agents can penetrate into cancer cells more effectively, thereby improving the ability of anticancer agents to kill cancer cells. Therefore, urokinase is also a It is a good adjuvant therapy for cancer, and it has no antigenicity problem and can be used for a long time. Urokinase is an injection drug, and the finished product has strict requirements on endotoxin. The Pharmacop...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/72B01J47/02
CPCC12N9/6462B01J47/02C12Y304/21073
Inventor 顾京李争倪萌蔡宏兵唐维
Owner JIANGSU YOULIKA BIOTECHNOLOYG CO LTD
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