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Method for preparing blood coagulation factor ⅷ, fibrinogen and fibronectin by cryoprecipitation

A technology of fibronectin and fibrinogen, which is applied to the preparation methods of fibrinogen and peptides, coagulation/fibrinolytic factors, etc., can solve the problems of insufficient utilization, achieve small batch differences, improve utilization, The effect of high yield and purity

Active Publication Date: 2011-12-28
SHANDONG TAIBANG BIOLOGICAL PROD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of preparation technology obviously cannot make full use of various useful components in the cryoprecipitate, and realize the comprehensive development and utilization of various plasma proteins in the cryoprecipitate.

Method used

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  • Method for preparing blood coagulation factor ⅷ, fibrinogen and fibronectin by cryoprecipitation
  • Method for preparing blood coagulation factor ⅷ, fibrinogen and fibronectin by cryoprecipitation
  • Method for preparing blood coagulation factor ⅷ, fibrinogen and fibronectin by cryoprecipitation

Examples

Experimental program
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Effect test

example 1

[0041] Step 1: Take 10kg of cryoprecipitate, add 30kg of water for injection at 30°C, stir and dissolve for 1 hour. Add 0.1mol / L acetic acid by spraying at a speed of 120ml / min, adjust the pH value to 6.9, adjust the conductivity to less than 4 mS / cm, and use about 1.4L to remove acetic acid. Add the well-balanced DEAE Sephadex A50 gel, stir and adsorb for 40 minutes, use a continuous flow centrifuge (tube centrifuge) to remove the gel and a small amount of undissolved precipitate, and collect the supernatant.

[0042] The following preparations should be made before the first step: Weigh 20g of DEAE Sephadex A50 gel (dried gel). Place the dry glue in a beaker, add 1L of 80mmol / L sodium chloride solution to swell for 6 hours. Prepare balance solution: 20mmol / L sodium citrate, 60mmol / L sodium chloride, pH 6.9. Discard the upper layer of swelling liquid in the beaker, keep the lower layer of gel, then add 1L of balance solution, stir evenly, and balance for 10-20 minutes, remo...

example 2

[0054] Step 1: Take 10kg of cryoprecipitate, add 50kg of water for injection at 35°C, stir and dissolve for 1 hour; add acetic acid to adjust the pH value to 6.7 by spraying at a speed of 100 ml / min. Add well-balanced DEAE Sephadex A50 gel, stir and absorb for 50 minutes;

[0055] The following preparations should be made before the first step: Weigh 15g of DEAE Sephadex A50 gel (dried gel). Prepare balance solution: 10mmol / L sodium citrate, 80mmol / L sodium chloride, pH 6.7;

[0056] All the other operating steps are the same as the first step of embodiment 1.

[0057] Step 2: Add 0.1 mol / L acetic acid by spraying continuously at a speed of 100 ml / min, and adjust the pH to 6.5. While adding acetic acid, cool down with an ice-water bath, and adjust the temperature to 12°C.

[0058] All the other operating steps are the same as the second step of Example 1.

[0059] The third step: collect 55 kg of the supernatant obtained in the second step, and the operation steps are the ...

Embodiment 3

[0076] Example 3: Research and testing of different adsorption methods and adsorption conditions

[0077] Many purification processes for coagulation factor VIII and fibrinogen include the step of aluminum hydroxide gel adsorption. Aluminum hydroxide gel can adsorb a variety of coagulation factors, but it is a non-specific adsorbent with limited adsorption capacity. Moreover, the preparation and quality control of aluminum hydroxide gel are more complicated and difficult to control. One of the main reasons why the existing technology cannot make good use of various active ingredients in the cryoprecipitate is that when the coagulation factors II, VII, IX, and X are not completely removed, due to the complicated extraction process of various proteins, The process and time-consuming are relatively long, and a fraction separated from the cryoprecipitate (such as a fraction rich in fibrinogen and fibronectin) will activate coagulation factors and coagulate when used to purify o...

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Abstract

The invention relates to the field of blood products, in particular to a process method for preparing three kinds of blood products from cryoprecitation, and aims to provide a complete separation and purification process for respectively and sequentially preparing the three kinds of blood products, i.e. a coagulation factor VIII, fibrinogen and fibronectin, from the cryoprecitation. The process method is implemented by the following technical scheme that the process method comprises the preparation steps of cryoprecitation dissolution, gel adsorption, precipitation of the fibrinogen and the fibronectin, two-step ion exchange chromatography and glycin / sodium chloride precipitation and inactivation of virus. The process method has the beneficial effects that the three kinds of useful blood products, i.e. the coagulation factor VIII, the fibrinogen and the fibronectin, can be separated and extracted from the cryoprecitation sequentially; and the utilization rate of blood plasma is greatly improved.

Description

technical field [0001] The invention relates to the field of blood products, in particular to a process capable of preparing three blood products from cryoprecipitate. technical background [0002] Cryoprecipitation is a precipitation component precipitated after fresh frozen plasma is thawed at low temperature, rich in coagulation factor VIII, fibrinogen and fibronectin. About half of the coagulation factor VIII and fibrinogen in the plasma, and most of the fibronectin will be enriched in the cryoprecipitate. [0003] Coagulation factor Ⅷ is the only specific drug for the treatment of hemophilia A, which has great clinical application value. Fibrinogen is the drug of choice for the treatment of congenital and acquired fibrinogen deficiency. Fibronectin also has high clinical value in the treatment of herpetic corneal ulcer. [0004] At present, almost all plasma-derived coagulation factor VIII in the world is prepared from cryoprecipitate; most of fibrinogen is prepared ...

Claims

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Application Information

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IPC IPC(8): C07K14/755C07K14/75C07K14/78C07K1/36C07K1/30C07K1/22C07K1/18
Inventor 师秀梅朱孟沼李斌菅长永席智赢邵玉娟郑志华张翠萍乔福锋孙永张谢忠兵
Owner SHANDONG TAIBANG BIOLOGICAL PROD CO LTD
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