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Preparation method of human coagulation factor VIII

A technology of human blood coagulation factor and plasma, which is applied in the field of preparation of human blood coagulation factor VIII, can solve problems such as production process optimization, and achieve the effect of saving resources and increasing efficiency

Active Publication Date: 2018-06-29
GUIZHOU TAIBANG BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In summary, the existing technology has not comprehensively optimized the production process. Therefore, it is imperative to optimize the preparation process of human blood coagulation factor VIII (FVIII) stock solution and improve the yield and purity of FVIII.

Method used

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  • Preparation method of human coagulation factor VIII
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  • Preparation method of human coagulation factor VIII

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] A preparation method for human blood coagulation factor VIII, comprising the steps of:

[0035] 1) Preparation of cryoprecipitate: prepare plasma cryoprecipitate;

[0036]2) Dissolving: cryoprecipitate is dissolved to obtain a solution;

[0037] 3) Acid precipitation: adjust the pH of the solution with acetic acid solution, and lower the temperature to 14°C, collect the supernatant by centrifugation after acid precipitation, the liquid temperature is 15°C, and the liquid output speed of the centrifuge is ≤5000ml / min;

[0038] 4) DEAE Sephadex A-50 gel adsorption: Add DEAE Sephadex A-50 gel to the supernatant obtained in step 3) for adsorption, the amount of A-50 gel is 0.0025 kg per kilogram of cryoprecipitate dry gel, A -50 dry glue soaked in swelling solution for more than 3 hours before use, and balanced with the balance solution for 3 times, each time not less than 10 minutes, the adsorption pH is 6.9, and the adsorption time is 60 minutes. The output speed of eac...

Embodiment 2

[0056] A preparation method for human blood coagulation factor VIII, comprising the steps of:

[0057] 1) Preparation of cryoprecipitate: prepare plasma cryoprecipitate;

[0058] 2) Dissolving: cryoprecipitate is dissolved to obtain a solution;

[0059] 3) Acid precipitation: adjust the pH of the solution with acetic acid solution, and lower the temperature to 12°C. After the acid precipitation, centrifuge to collect the supernatant, the liquid temperature is 10°C, and the liquid output speed of the centrifuge is ≤5000ml / min;

[0060] 4) DEAE Sephadex A-50 gel adsorption: add DEAE Sephadex A-50 gel to the supernatant obtained in step 3) for adsorption, the amount of A-50 gel is 0.0025k per kg of cryoprecipitate dry gel, A -50 dry glue soaked in swelling solution for more than 3 hours before use, and balanced with balance solution for 3 times, each time not less than 10 minutes, the adsorption pH is 6.9, and the adsorption time is 30-60 minutes. After adsorption, the supernata...

Embodiment 3

[0078] A preparation method for human blood coagulation factor VIII, comprising the steps of:

[0079] 1) Preparation of cryoprecipitate: prepare plasma cryoprecipitate;

[0080] 2) Dissolving: cryoprecipitate is dissolved to obtain a solution;

[0081] 3) Acid precipitation: adjust the pH of the solution with acetic acid solution, and lower the temperature to 16°C, collect the supernatant by centrifugation after acid precipitation, the liquid temperature is 20°C, and the liquid output speed of the centrifuge is ≤5000ml / min;

[0082] 4) DEAE Sephadex A-50 gel adsorption: add DEAE Sephadex A-50 gel to the supernatant obtained in step 3) for adsorption, the amount of A-50 gel is 0.0025 kg per kg of cryoprecipitate dry gel, A -50 dry glue is soaked in swelling solution for more than 3 hours before use, and balanced with balance solution for 3 times, each time not less than 10 minutes, the adsorption pH is 6.9, and the adsorption time is 30-60 minutes. After adsorption, the super...

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Abstract

The invention belongs to the technical field of biological pharmacy and blood products, and especially relates to a preparation method of a human coagulation factor VIII. According to the invention, apreparation process of the human coagulation factor VIII is subjected to total research, cryoprecitation preparation, dissolution, washing liquid and an eluate are screened, after elution, the residual quantity of sorbate 80 and tributyl phosphate can achieve the requirement of current pharmacopeia; a dialysate is screened, the finally obtained product has the advantages of no collapse, no atrophy, and fast redissolving time, and FVIII titer after dry heat inactivation is 28.9 IU / ml. The processes for preparing the human coagulation factor VIII are optimized and screened, an optimum scheme isobtained, the recovery rate of each process is calculated, the recovery rate of FVIII is 28.91%, the efficiency for preparing FVIII by blood plasma is effectively increased, and a rear blood plasma resource is indirectly saved.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals and blood products, and in particular relates to a preparation method of human blood coagulation factor VIII. Background technique [0002] Human coagulation factor VIII (FVIII) is a plasma protein product mainly containing FⅧ, which is isolated from healthy human plasma, and is mainly used for the treatment of hemophilia A (hemophilia A, HA). According to the World Health Organization (WHO) and World Federation of Hemophilia (World Federation of Hemophilia, WFH) reports, the global incidence of HA is 15-20 / 100,000, and China is about 2.27-2.84 / 100,000. For HA patients, FⅧ infusion is required for life To maintain normal life (Liang Xiaoming, Huang Pan, Zhou Xing, et al. Research on the extraction process of human coagulation factor Ⅷ [J]. Anhui Agricultural Sciences, 2013, 41(30): 12043-12044, 12048.). [0003] At present, the FⅧ market is still in a tense situation. Because the conte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/755C07K1/36C07K1/34C07K1/30C07K1/16
CPCC07K14/755
Inventor 陈云华陈艾军向宏王勇符平松赵学梅刘欣晏杨刚
Owner GUIZHOU TAIBANG BIOLOGICAL PROD
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