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Dry heat treatment stabilizer for human blood coagulation factor VIII and application thereof

A technology of human coagulation factor and dry heat treatment, which is applied in the direction of coagulation/fibrinolytic factor, enzyme stabilization, factor VII, etc., can solve the problems of low activity recovery rate of human coagulation factor VIII, low application value, and low cost, and achieve good clinical results Application prospects, low cost, and few types of effects

Active Publication Date: 2014-07-23
CHENGDU RONGSHENG PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The stabilizer has only two components, does not need to use a large amount of human serum albumin, and is low in cost, but the activity recovery rate of human coagulation factor VIII is the lowest, and its practical application value is low

Method used

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  • Dry heat treatment stabilizer for human blood coagulation factor VIII and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1 Preparation of human coagulation factor VIII preparation of the present invention

[0023] 1. Preparation method

[0024] Ⅰ. Separation and purification

[0025] (1) Using fresh frozen plasma as raw material, melt the plasma and centrifuge to prepare cryoprecipitate, dissolve the cryoprecipitate in 0.02M Tris buffer, precipitate with 30% polyethylene glycol, absorb with aluminum hydroxide and centrifuge to obtain the supernatant;

[0026] (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 inactivation);

[0027] (3) Use Toyopearl DEAE 650M as the gel for further purification by ion exchange chromatography, the buffer is 0.001M sodium citrate buffer, by changing the sodium chloride ionic strength of the chromatography buffer , and the eluate containing hu...

Embodiment 2

[0031] Embodiment 2 Preparation of human coagulation factor VIII preparation of the present invention

[0032] 1. Preparation method

[0033] Ⅰ. Separation and purification

[0034] (1) Using fresh frozen plasma as raw material, melt the plasma and centrifuge to prepare cryoprecipitate, dissolve the cryoprecipitate in 0.02M Tris buffer, precipitate with 30% polyethylene glycol, absorb with aluminum hydroxide and centrifuge to obtain the supernatant;

[0035] (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 inactivation);

[0036] (3) Using Toyopearl DEAE 650M as the gel as the filler for further purification by ion exchange chromatography, the buffer is 0.05M sodium citrate buffer, and collected by changing the sodium chloride ionic strength of the chromatography buffer E...

Embodiment 3

[0040] Embodiment 3 Preparation of human coagulation factor VIII preparation of the present invention

[0041] 1. Preparation method

[0042] Ⅰ. Separation and purification

[0043] (1) Using fresh frozen plasma as raw material, melt the plasma and centrifuge to prepare cryoprecipitate, dissolve the cryoprecipitate in 0.02M Tris buffer, precipitate with 30% polyethylene glycol, absorb with aluminum hydroxide and centrifuge to obtain the supernatant;

[0044] (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 inactivation);

[0045] (3) Use Toyopearl DEAE 650M as the gel as the filler for further purification by ion exchange chromatography, the buffer is 0.02M sodium citrate buffer, and the sodium chloride ionic strength of the chromatography buffer is changed to collect and ...

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Abstract

Provided is a dry heat treatment stabilizer for human blood coagulation factor VIII, which comprises the following proportions of components: 0.005 M-0.015 M of sodium citrate, 0.001 M-0.002 M of calcium chloride, 0. 15 M-0.25 M of arginine hydrochloride, and 6-10 g / L of human serum albumin. Also provided are a human blood coagulation factor VIII preparation and a preparation method therefor. The dry heat treatment stabilizer for human blood coagulation factor VIII can maintain the activity of human blood coagulation factor VIII.

Description

technical field [0001] The invention relates to the field of plasma products, in particular to a dry heat treatment stabilizer for human blood coagulation factor VIII and its application. Background technique [0002] Human blood coagulation factor Ⅷ is a kind of coagulation factor, which is a composite factor consisting of two parts, of which the small molecular weight (about 260KD) part Ⅷ: C has procoagulant activity and acts as FIXa in the cascade reaction of blood coagulation A cofactor that activates FX into FXa, which can correct the coagulation abnormalities of hemophilia A. The preparation of human coagulation factor VIII from plasma is the main human coagulation factor VIII product currently on the market. Common methods for the preparation of human blood coagulation factor VIII include precipitation and chromatography. Precipitation includes isoelectric point precipitation, salting out precipitation, ether precipitation, acid precipitation and PEG precipitation. C...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/00
CPCC07K14/755C07K1/30C07K1/36A61K38/37A61K9/00C07K1/18C12N9/96
Inventor 牟蕾鲁涛王黔川李伟余伟
Owner CHENGDU RONGSHENG PHARMA
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