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Method for preparing cryoprecipitate and method for preparing blood coagulation factor VIII preparation by using cryoprecipitate

A technology of cryoprecipitation and polyethylene glycol precipitation, which is applied in the preparation method of peptides, coagulation/fibrinolytic factors, factor VII, etc., can solve the problems of waste of plasma resources and less cryoprecipitation, and achieve full utilization and good economic benefits , the effect of good market application prospects

Active Publication Date: 2014-06-11
CHENGDU RONGSHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the amount of cryoprecipitate produced by this method is relatively small, usually only 8.9kg of cryoprecipitate is obtained per ton of plasma, and the content of human coagulation factor VIII in the cryoprecipitate is 39.951IU / g (see battalion chief Yong et al., "Human coagulation factor VIII Separation and Purification Technology Research", Master's Thesis of Shandong University), the amount of human blood coagulation factor Ⅷ preparation prepared by further separation and purification of cryoprecipitate is also relatively small, resulting in a waste of plasma resources

Method used

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  • Method for preparing cryoprecipitate and method for preparing blood coagulation factor VIII preparation by using cryoprecipitate
  • Method for preparing cryoprecipitate and method for preparing blood coagulation factor VIII preparation by using cryoprecipitate
  • Method for preparing cryoprecipitate and method for preparing blood coagulation factor VIII preparation by using cryoprecipitate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 Prepare cryoprecipitate with the method of the present invention

[0033] 1. Experimental equipment

[0034] Weir filter: model is FL-2023 weir tank filter;

[0035] Continuous centrifuge: GQ142 high-speed tube centrifuge.

[0036] 2. Experimental method

[0037] (1) After collection of fresh healthy human plasma, store it at -20°C for no more than 3 years;

[0038] (2) Pre-thawing: put 2500L of fresh frozen plasma (2575Kg, the content of human coagulation factor VIII is 2500,000IU), place it at an ambient temperature of 0°C, and heat it up to -10°C to 0°C;

[0039] (3) Melting: melt in a water bath at 25°C, and heat the plasma to 0-5°C to melt the plasma;

[0040] (4) Filtration: Under the condition of maintaining the temperature of the melted plasma at 0-5°C, filter with a weir filter to obtain filtrate and filter residue;

[0041] (5) Centrifugation: Under the condition of maintaining the temperature of the filtrate at 0-5°C, use a continuous centri...

Embodiment 2

[0047] Embodiment 2 Prepare cryoprecipitate with the method of the present invention

[0048] 1. Experimental equipment

[0049] Weir filter: model is FL-2023 weir tank filter;

[0050] Continuous centrifuge: GQ142 high-speed tube centrifuge.

[0051] 2. Experimental method

[0052] (1) After collection of fresh healthy human plasma, store it at -20°C for no more than 3 years;

[0053] (2) Pre-thawing: put 2500L of fresh frozen plasma (2575Kg, the content of human blood coagulation factor VIII is 2500,000IU), place it at an ambient temperature of 2°C, and heat it up to -10°C to 0°C;

[0054] (3) Melting: melt in a water bath at 37°C, and heat the plasma to 0-5°C to melt the plasma;

[0055] (4) Filtration: Under the condition of maintaining the temperature of the melted plasma at 0-5°C, filter with a weir filter to obtain filtrate and filter residue;

[0056] (5) Centrifugation: Under the condition of maintaining the temperature of the filtrate at 0-5°C, use a continuous ...

Embodiment 3

[0058] Example 3 Preparation of human blood coagulation factor VIII preparation by cryoprecipitation of the present invention

[0059] 1. Experimental method

[0060] 300g of the cryoprecipitate (filter residue) obtained by filtration in step (4) of Example 1 (repeated three times) and the cryoprecipitate (precipitate) obtained by centrifugation in step (5) of Example 1 (the weights are 3.8kg, 3.525kg, 2kg respectively) according to The following method is used to purify and prepare human coagulation factor VIII:

[0061] (1) Dissolve the cryoprecipitate with 0.02M tromethamine (Tris) buffer, precipitate with 30% polyethylene glycol, centrifuge, and obtain the supernatant;

[0062] (2) After the supernatants were combined and clarified, Tween-80 and tributyl phosphate were added to make the final concentrations 1% and 0.3%, respectively, and treated at 25°C±1°C for 6 hours to complete the first virus inactivation (that is, SD virus inactivated);

[0063] (3) Sodium chloride...

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Abstract

The invention discloses a method for preparing a cryoprecipitate. The method comprises the steps of (1) thawing: selecting fresh frozen plasma, and heating to obtain thawed plasma of which the temperature is 0-5 DEG C; (2) filtering: filtering under the condition that the temperature of the thawed plasma is 0-5 DEG C to obtain filtrate and filter residues; (3) centrifuging: centrifuging under the condition that the temperature of the filtrate is 0-5 DEG C to obtain a precipitate; (4) combining the filter residues obtained in step (2) and the precipitate obtained in step (3) to obtain the cryoprecipitate. The preparation method disclosed by the invention is simple, and the prepared cryoprecipitate is high in yield and human blood coagulation factor VIII content, so the preparation method has a good industrial application prospect.

Description

technical field [0001] The invention relates to a method for preparing blood products, in particular to a method for preparing cryoprecipitate and a method for preparing blood coagulation factor VIII by using it. Background technique [0002] Cryoprecipitate is a white precipitate that is insoluble in fresh frozen plasma under low temperature conditions. Raw material for blood coagulation factor VIII preparations. [0003] Currently, there are two methods for plasma cryoprecipitation: rapid thawing centrifugation and siphoning. In the cryoprecipitate prepared by the siphon method, the content of human coagulation factor VIII is low, and the rapid melting centrifugation method is usually used at present. [0004] The specific steps of the rapid thawing centrifugation method are as follows: take out the fresh frozen plasma to be prepared for cryoprecipitate, put it in a refrigerator at 4±2°C to thaw overnight or melt it in a water bath at 4±2°C; Centrifuge at ±2°C, and the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K14/755C07K1/36C07K1/30C07K1/18
Inventor 牟蕾鲁涛初毅波苗松邓红王黔川李伟余伟
Owner CHENGDU RONGSHENG PHARMA
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