Preparation process for extracting human fibrinogen from component I

Abstract

The invention relates to a preparation process for extracting human fibrinogen from a component I, and belongs to the technical field of blood products. The process comprises the following steps: dissolving an FI precipitate in a dissolving solution A containing sodium citrate, lysine hydrochloride and heparin sodium, and filtering to obtain a filtrate A; adding PEG (Polyethylene Glycol) into the filtrate A until the final concentration is 5%, purifying and separating, collecting PEG precipitate, dissolving and filtering to obtain filtrate B; adding polysorbate 80 and tributyl phosphate into the filtrate B to carry out S / D inactivation; cooling the inactivated liquid, adding an ethanol solution for purification and separation, collecting the precipitate, and dissolving to obtain a filtrate C; adding glycine into the filtrate C until the final concentration is 1.8 mol / L, performing purification and separation, collecting precipitates, and dissolving to obtain filtrate D; performing ultrafiltration dialysis on the filtrate D to obtain a stock solution; and diluting the stock solution to a proper concentration, sterilizing, filtering, sub-packaging, freeze-drying, and carrying out dry heat inactivation treatment to obtain a finished product. The method is simple in process and low in cost, and the prepared human fibrinogen is high in purity, good in safety and suitable for clinical hemostasis and tissue repair.

Description

Technical Field

[0001] This invention belongs to the field of blood products technology, specifically relating to a preparation process for extracting human fibrinogen from component I. Background Technology

[0002] Human fibrinogen (Fg) is a protein component in blood plasma, with a concentration as high as 2-4 g / L. It plays a crucial role in the coagulation system. The final stage of coagulation involves fibrinogen being converted into fibrin monomers by thrombin, which then cross-link with factor XIII to form an insoluble fibrin clot, achieving hemostasis. Patients with congenital fibrinogen deficiency or acquired fibrinogen deficiency due to disease are prone to coagulation dysfunction, and fibrinogen supplementation is currently the only reliable treatment.

[0003] Human fibrinogen is a blood product with national standards, included in the 2025 edition of the Chinese Pharmacopoeia, Volume III. This drug has been used clinically in China for many years. Currently, human fibrinogen can be produced on a large scale both domestically and internationally, but the yield and quality vary greatly, making it difficult to simultaneously achieve high yield and high quality. Analysis of existing human fibrinogen preparation processes reveals that domestic manufacturers with drug approval mainly use low-temperature ethanol processes and ion exchange methods. However, these methods involve the activation and degradation of fibrinogen during protein purification, and the presence of a certain amount of fibronectin, resulting in low purity of the currently produced products. Foreign manufacturers use processes such as aluminum hydroxide adsorption, ion exchange, and glycine purification, which improves purity but is more complex. Summary of the Invention

[0004] The purpose of this invention is to design a preparation process for extracting human fibrinogen from component I. In view of the current production status of human fibrinogen, this invention aims to develop an advanced preparation process to obtain a fibrinogen product with high yield, high purity, resistance to dry heat, and rapid solubility.

[0005] The technical solution adopted by this invention to solve its technical problem is: a preparation process for extracting human fibrinogen from component I, comprising the following steps:

[0006] (1) Dissolution and filtration of FI precipitate: Add dissolving solution A at 10 to 12 times the weight of FI precipitate, control the dissolution temperature at 20 to 30°C, and collect filtrate A after dissolving for 0.5 to 2 hours;

[0007] (2) PEG purification and separation: According to the weight of filtrate A in step (1), add PEG to a final concentration of 5%, stir at 10-20℃ for 0.5-1h, centrifuge, and collect the PEG precipitate; add dissolving solution B at 10-15 times the weight of PEG precipitate, dissolve at 20-30℃ for 1-2 hours, filter, and collect filtrate B.

[0008] (3) S / D inactivation: Add S / D reagent to filtrate B in step (2) so that the final content of polysorbate 80 in filtrate B is 1% and the final content of tributyl phosphate is 0.3%. Inactivate at 24-26℃ for no less than 6 hours to obtain inactivation solution;

[0009] (4) Ethanol purification and separation: When the inactivation solution in step (3) is cooled to 0-2℃, slowly add 50% ethanol solution below -15℃ until the ethanol content of the inactivation solution is 8%. After adding ethanol, cool the inactivation solution to -2.5-0℃, stir for 40-60 min and centrifuge to collect the ethanol precipitate. Dissolve the ethanol precipitate with 12-17 times the amount of dissolving solution B at 20-30℃ for 1-2 hours, filter and collect the filtrate C.

[0010] (5) Glycine purification and separation: Add glycine to the filtrate C in step (4) to a final concentration of 1.8 mol / L. After it is fully dissolved, cool the solution to 2-10℃, stir for at least 60 min, centrifuge, and collect the precipitate. Dissolve the glycine precipitate in 4-8 times the amount of dissolving solution C. After dissolving at room temperature for 1-2 h, filter and collect the filtrate D.

[0011] (6) Ultrafiltration dialysis: Dialyze the filtrate D from step (5) with a constant weight of 3 to 6 times using the dissolving solution C, and then concentrate it by ultrafiltration to a protein concentration of 30 g / L or higher, which is the original solution;

[0012] (7) Preparation: Calculate the final volume based on the protein content and packaging specifications of the stock solution in step (6), dilute the stock solution to 23-28 g / L with dialysis buffer, stir evenly, and confirm the pH value of the product to be 6.8-7.2; filter the prepared product with a sterile filter cartridge to obtain the semi-finished product;

[0013] (8) Finished product: The semi-finished product is packaged and freeze-dried. The freeze-dried product is then capped and sealed, and then subjected to dry heat inactivation treatment at 100±1℃ for 30 minutes to obtain the finished product.

[0014] Preferably, the FI precipitate in step (1) is freshly prepared or thawed component I. Frozen component I is first chopped into small cubes of about 1 cm, then dissolved, and filtered with a 30SP filter cartridge to obtain filtrate A.

[0015] Preferably, the formulation of solution A is as follows: each kilogram of water for injection contains 29.4 g of sodium citrate, 9.1 g of lysine hydrochloride, 6000 IU of sodium heparin, and a pH value of 6.8–7.0.

[0016] Preferably, the formulation of solution B is as follows: per kilogram of water for injection, there are 15g of sodium citrate, 9g of sodium chloride, 3.5g of lysine hydrochloride, 10g of sucrose, and the pH value is 6.8-7.2.

[0017] Preferably, the formulation of solution C is as follows: each kilogram of water for injection contains 10g of sodium citrate, 2g of sodium glutamate, 20g of arginine hydrochloride, and a pH value of 6.8–7.2.

[0018] Preferably, the S / D reagent formulation includes: 11% polysorbate 80 and 3.3% tributyl phosphate.

[0019] Preferably, the specific steps of step (2) are as follows: adjust the filtrate A from step (1) to 6.8-7.2 and then add PEG. After stirring, centrifuge with a maximum processing capacity of ≤2.0Kg per minute. Control the temperature of the centrifuged liquid at 10-17℃. Filter the dissolved PEG precipitate with a 60SP filter cartridge to obtain filtrate B.

[0020] Preferably, step (3) is as follows: adjust the pH of the filtrate B from step (2) to 6.8-7.2, add S / D reagent at 0.1 times the weight of the product, stir evenly and filter into the S / D inactivation tank, heat the product to 24-26°C and perform S / D virus inactivation for at least 6 hours.

[0021] Preferably, further, step (4) is as follows: adjust the pH of the inactivation solution in step (3) to 6.8 to 7.2, add ethanol after it cools down, stir and centrifuge, with a maximum processing capacity of ≤2.0Kg per minute, and control the temperature of the centrifuged liquid outlet at -2.5 to 0℃. The dissolved ethanol precipitate is filtered with a 60SP filter element to obtain filtrate C.

[0022] Preferably, step (5) is as follows: adjust the pH of the filtrate C from step (4) to 7.1-7.3, add glycine, stir and centrifuge, with a maximum processing capacity of ≤2.0Kg per minute, and control the temperature of the centrifuged liquid at 0-4℃. The dissolved glycine precipitate is filtered with a 0.45μm filter element to obtain filtrate D.

[0023] The beneficial effects of this invention are:

[0024] 1. This invention employs three different precipitation methods (PEG precipitation, ethanol precipitation, and glycine precipitation) to separate and purify human fibrinogen, achieving a high fibrinogen yield of over 1600 vials (0.5g / vial) per ton of plasma, significantly exceeding the level of existing technologies.

[0025] 2. The key quality indicators of the fibrinogen product prepared by this invention are superior to those of commercially available products, with a purity of 91-98%, a coagulation activity of 20-23 seconds, and a short reconstitution time of only 2-5 minutes, which can better meet the needs of clinical applications.

[0026] 3. The precipitation method used in this invention requires less equipment investment, is simple to implement, and is conducive to commercial production. It solves the problem that the preparation process for extracting human fibrinogen from component I in the prior art is difficult to commercialize. Attached Figure Description

[0027] Figure 1 This is a process flow diagram of the present invention. Detailed Implementation

[0028] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings and embodiments. Obviously, the described embodiments are merely some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0029] Example 1

[0030] (1) Dissolution and filtration of FI precipitate

[0031] Chop the 5 kg of frozen FI precipitate into small cubes of about 1 cm, add dissolving solution A at 10 times the weight of the FI precipitate, control the temperature of the dissolving solution at 25.0℃, and filter after dissolving for 2 hours.

[0032] Filter using a 30SP filter cartridge and collect filtrate A.

[0033] (2) Purification and separation of polyethylene glycol (PEG)

[0034] Measure the pH of filtrate A and adjust it to 7.07. According to the weight of filtrate A in step (1), add 11.7 kg of PEG solution, and at the same time cool filtrate A to 10.2℃, stir for 1 h and then centrifuge.

[0035] The maximum processing capacity is 1.5 kg per minute. The centrifuged liquid temperature is controlled at 12-14℃, and the PEG precipitate is collected.

[0036] Add dissolving solution B at 10 times the weight of the PEG precipitate, dissolve at 25.8°C for 2 hours, and then filter.

[0037] The dissolved PEG precipitate was filtered using a 60SP filter cartridge, and the filtrate B was collected for S / D inactivation.

[0038] (3) S / D inactivation

[0039] Measure the pH of filtrate B and adjust it to 6.8–7.2. Add 4.1 kg of the prepared S / D reagent according to the weight of filtrate B to make the final content of polysorbate 80 in the product 1% and the final content of tributyl phosphate 0.3%. Stir well and filter into the S / D inactivation tank.

[0040] The product was heated to 24–26°C for S / D virus inactivation, and the inactivation was carried out continuously for 6 hours and 8 minutes.

[0041] (4) Ethanol purification and separation

[0042] Adjust the pH of the inactivated product to 7.05, and when it cools down to -2.0℃, slowly add 7.9 kg of 50% ethanol solution below -15℃. After adding the ethanol, cool the product down to -2.0℃, stir for 60 min, and then centrifuge.

[0043] The maximum processing capacity is 1.3 kg per minute, the centrifuged liquid outlet temperature is controlled at -1.0℃, and the ethanol precipitate is collected.

[0044] The ethanol precipitate was dissolved in 15 times the amount of solution B at 24.9°C for 2 hours and then filtered.

[0045] The dissolved ethanol precipitate was filtered using a 60SP filter cartridge, and the filtrate C was collected for glycine purification.

[0046] (5) Glycine purification and separation

[0047] Measure the pH of filtrate C and adjust it to 7.18. Add 6.27 kg of glycine to filtrate C in step (4) and let it dissolve completely. Then cool the solution to 3.8°C, stir for at least 60 min, and centrifuge.

[0048] The maximum processing capacity is 1.0 kg per minute. The temperature of the centrifuged liquid is controlled at 2-3℃, and the glycine precipitate is collected.

[0049] The glycine precipitate was dissolved in 6 times the amount of solution C, and filtered after dissolving at room temperature for 2 hours.

[0050] Filter the solution using a 0.45μm filter cartridge to obtain filtrate D.

[0051] (6) Ultrafiltration dialysis

[0052] After concentrating filtrate D to 3 to 6 times the weight of glycine precipitate, 41.15 kg of dissolving solution C was used to perform constant weight dialysis on filtrate D from step (5), and ultrafiltration was used to concentrate it to a protein concentration of 31 g / L, which is the original solution.

[0053] (7) Preparation

[0054] Calculate the final volume based on the protein content and packaging specifications of the stock solution in step (6), dilute the stock solution to 26.5 g / L with dialysis buffer, stir evenly, and confirm the pH value of the product as 6.8 to 7.2.

[0055] The prepared product is sterilized and filtered using a 0.22μm sterilization filter cartridge to obtain a semi-finished product.

[0056] (8) Finished product

[0057] After the semi-finished products are packaged, they are freeze-dried. The temperature of the products during the freeze-drying process must not exceed 35°C, and they are then vacuum-sealed.

[0058] After freeze-drying, the product is transferred to the capping room under aseptic conditions for capping and sealing.

[0059] After capping, the product is transferred to a water bath sterilizer for continuous dry heat inactivation at 100±1℃ for 30 minutes to obtain the finished product.

[0060] Example 2

[0061] (1) Dissolution and filtration of FI precipitate

[0062] Chop the 5 kg of frozen FI precipitate into small cubes of about 1 cm, add dissolving solution A at 10 times the weight of the FI precipitate, control the temperature of the dissolving solution at 26.3℃, and filter after dissolving for 2 hours.

[0063] Filter using a 30SP filter cartridge and collect filtrate A.

[0064] (2) Purification and separation of polyethylene glycol (PEG)

[0065] Measure the pH of filtrate A and adjust it to 6.99. According to the weight of filtrate A in step (1), add 11.6 kg of PEG solution, and at the same time cool filtrate A to 10.1℃, stir for 1 h and then centrifuge.

[0066] The maximum processing capacity is 1.6 kg per minute. The centrifugal outlet temperature is controlled at 15-16℃, and the PEG precipitate is collected.

[0067] Add 10 times the weight of the PEG precipitate to solution B, dissolve at 26.4℃ for 2 hours, and then filter.

[0068] The dissolved PEG precipitate was filtered using a 60SP filter cartridge, and the filtrate B was collected for S / D inactivation.

[0069] (3) S / D inactivation

[0070] Measure the pH of filtrate B and adjust it to 6.8–7.2. Add 4.2 kg of the prepared S / D reagent according to the weight of the product, so that the final content of polysorbate 80 in the product is 1% and the final content of tributyl phosphate is 0.3%. After stirring evenly, filter into the S / D inactivation tank.

[0071] The product was heated to 24-26°C to inactivate the S / D virus for 6 hours and 5 minutes.

[0072] (4) Ethanol purification and separation

[0073] Adjust the pH of the inactivated product to 7.00, and when it cools down to -1.6℃, slowly add 7.9 kg of 50% ethanol solution below -15℃. After adding the ethanol, cool the product down to -1.7℃, stir for 60 min, and then centrifuge.

[0074] The maximum processing capacity is 1.5 kg per minute, the centrifuged liquid outlet temperature is controlled at -1.0℃, and the ethanol precipitate is collected.

[0075] The ethanol precipitate was dissolved in 15 times the amount of solution B at 24.9°C for 2 hours and then filtered.

[0076] The dissolved ethanol precipitate was filtered using a 60SP filter cartridge, and the filtrate C was collected for glycine purification.

[0077] (5) Glycine purification and separation

[0078] Measure the pH of filtrate C and adjust it to 7.18. Add 6.27 kg of glycine to filtrate C in step (4) and let it dissolve completely. Then cool the solution to 2.6°C, stir for at least 60 min, and centrifuge.

[0079] The maximum processing capacity is 1.4 kg per minute. The centrifugal outlet temperature is controlled at 3℃, and the glycine precipitate is collected.

[0080] The glycine precipitate was dissolved in 6 times the amount of dissolving solution C, and filtered after dissolving at room temperature for 2 hours.

[0081] Filter the solution using a 0.45μm filter cartridge to obtain filtrate D.

[0082] (6) Ultrafiltration dialysis

[0083] After concentrating filtrate D to 3 to 6 times the weight of glycine precipitate, 41.1 kg of dissolving solution C was used to perform constant weight dialysis on filtrate D from step (5), and ultrafiltration was used to concentrate it to a protein concentration of 35 g / L, which is the original solution.

[0084] (7) Preparation

[0085] Calculate the final volume based on the protein content and packaging specifications of the stock solution in step (6), dilute the stock solution to 26.0 g / L with dialysis buffer, stir evenly, and confirm the pH value of the product as 6.8 to 7.2.

[0086] The prepared product is sterilized and filtered using a 0.22μm sterilization filter cartridge to obtain a semi-finished product.

[0087] (8) Finished product

[0088] After the semi-finished products are packaged, they are freeze-dried. The temperature of the products during the freeze-drying process must not exceed 35°C, and they are then vacuum-sealed.

[0089] After freeze-drying, the product is transferred to the capping room under aseptic conditions for capping and sealing.

[0090] After capping, the product is transferred to a water bath sterilizer for continuous dry heat inactivation at 100±1℃ for 30 minutes to obtain the finished product.

[0091] Example 3

[0092] (1) Dissolution and filtration of FI precipitate

[0093] Chop the 5 kg of frozen FI precipitate into small cubes of about 1 cm, add dissolving solution A at 10 times the weight of the FI precipitate, control the temperature of the dissolving solution at 25.0℃, and filter after dissolving for 2 hours.

[0094] Filter using a 30SP filter cartridge and collect filtrate A.

[0095] (2) Purification and separation of polyethylene glycol (PEG)

[0096] Measure the pH of filtrate A and adjust it to 6.97. According to the weight of filtrate A in step (1), add 11.4 kg of PEG solution, and at the same time cool the filtrate to 10.1℃, stir for 1 h and then centrifuge.

[0097] The maximum processing capacity is 1.7 kg per minute. The centrifuged liquid temperature is controlled at 12-14℃, and the PEG precipitate is collected.

[0098] Add 10 times the weight of the PEG precipitate to solution B, dissolve at 26.9°C for 2 hours, and then filter.

[0099] The dissolved PEG precipitate was filtered using a 60SP filter cartridge, and the filtrate B was collected for S / D inactivation.

[0100] (3) S / D inactivation

[0101] Measure the pH of filtrate B and adjust it to 6.8–7.2. Add 4.6 kg of the prepared S / D reagent according to the weight of the product, so that the final content of polysorbate 80 in the product is 1% and the final content of tributyl phosphate is 0.3%. After stirring evenly, filter into the S / D inactivation tank.

[0102] The product was heated to 24–26°C to inactivate the S / D virus for 6 hours and 6 minutes.

[0103] (4) Ethanol purification and separation

[0104] Adjust the pH of the inactivated product to 7.00, and when it cools down to -1.3℃, slowly add 8.7 kg of 50% ethanol solution below -15℃. After adding the ethanol, cool the product down to -1.6℃, stir for 60 min, and then centrifuge.

[0105] The maximum processing capacity is 1.5 kg per minute, the centrifuged liquid outlet temperature is controlled at -1.0℃, and the ethanol precipitate is collected.

[0106] The ethanol precipitate was dissolved in 15 times its volume of solution B at 25.0°C for 2 hours and then filtered.

[0107] The dissolved ethanol precipitate was filtered using a 60SP filter cartridge, and the filtrate C was collected for glycine purification.

[0108] (5) Glycine purification and separation

[0109] Measure the pH of filtrate C and adjust it to 7.20. Add 6.24 kg of glycine to filtrate C in step (4) and let it dissolve completely. Then cool the solution to 2.7°C, stir for at least 60 min, and centrifuge.

[0110] The maximum processing capacity is 1.4 kg per minute. The centrifugal outlet temperature is controlled at 1℃, and glycine precipitate is collected.

[0111] The glycine precipitate was dissolved in 6 times the amount of solution C, and then filtered after dissolving at 25.2℃ for 2 hours.

[0112] Filter the solution using a 0.45μm filter cartridge to obtain filtrate D.

[0113] (6) Ultrafiltration dialysis

[0114] After concentrating filtrate D to 3 to 6 times the weight of glycine precipitate, 42.9 kg of dissolving solution C was used to perform constant weight dialysis on filtrate D from step (5), and ultrafiltration was used to concentrate it to a protein concentration of 40 g / L, which is the original solution.

[0115] (7) Preparation

[0116] Calculate the final volume based on the protein content and packaging specifications of the stock solution in step (6), dilute the stock solution to 26.0 g / L with dialysis buffer, stir evenly, and confirm the pH value of the product as 6.8 to 7.2.

[0117] The prepared product is sterilized and filtered using a 0.22μm sterilization filter cartridge to obtain a semi-finished product.

[0118] (8) Finished product

[0119] After the semi-finished products are packaged, they are freeze-dried. The temperature of the products during the freeze-drying process must not exceed 35°C, and they are then vacuum-sealed.

[0120] After freeze-drying, the product is transferred to the capping room under aseptic conditions for capping and sealing.

[0121] After capping, the product is transferred to a water bath sterilizer for continuous dry heat inactivation at 100±1℃ for 30 minutes to obtain the finished product.

[0122] The test results of the human fibrinogen products prepared in Examples 1-3 are as follows: sample Reconstitution time (min) Visible foreign objects purity(%) Solidification vitality (S) Total Fg (g) Example 1 5 The solution was clear with a slight opalescence. 91.0 20 0.6 Example 2 3 The solution was clear with a slight opalescence. 97.6 22 0.6 Example 3 2 The solution was clear with a slight opalescence. 98.2 23 0.6 Pharmacopoeia Standard ≤30 A small amount of flocculent material or protein particles is permissible. ≥70 ≤60 0.5

[0123] This process uses fibrinogen (FI) as raw material to separate and purify it. It employs three precipitation methods: PEG precipitation, ethanol precipitation, and glycine precipitation, combined with S / D method, freeze drying, and dry heat method for virus inactivation. Compared with fibrinogen separated by cryoprecipitation, it has a higher comprehensive utilization rate of plasma and a product yield of 1600 bottles / plasma. Compared with chromatographic separation and purification methods, large-scale production requires less investment in equipment, only centrifuges are needed, which can save on the cost of chromatographic equipment and chromatographic packing materials.

Claims

1. A process for the preparation of human fibrinogen from Fraction I, characterized in that, The method comprises the following steps: (1) Dissolution and filtration of the FI precipitate Add 10-12 times the weight of the FI precipitate with dissolving solution A, control the temperature at 20-30℃, and collect the filtrate A after 0.5-2 hours of dissolution; (2) PEG purification and separation According to the weight of the filtrate A in step (1), add PEG to a final concentration of 5%, stir for 0.5-1 hour at 10-20℃, and collect the PEG precipitate after centrifugation; add 10-15 times the weight of the PEG precipitate with dissolving solution B, and collect the filtrate B after 1-2 hours of dissolution at 20-30℃ and filtration; (3) S / D inactivation Add S / D reagent to the filtrate B in step (2) to make the final content of polysorbate 80 in the filtrate B 1% and the final content of tributyl phosphate 0.3%, and inactivate the mixture at 24-26℃ for not less than 6 hours to obtain an inactivated solution; (4) Ethanol purification and separation Slowly add 50% ethanol solution below -15℃ to the inactivated solution in step (3) to make the ethanol content 8%, cool the inactivated solution to 0-2℃, and collect the ethanol precipitate after stirring for 40-60 minutes and centrifugation; dissolve the ethanol precipitate with 12-17 times the weight of dissolving solution B at 20-30℃, and collect the filtrate C after filtration; (5) Glycine purification and separation Add glycine to the filtrate C in step (4) to a final concentration of 1.8 mol / L, cool the solution to 2-10℃, and collect the precipitate after stirring for at least 60 minutes and centrifugation; dissolve the precipitate with 4-8 times the weight of dissolving solution C, and collect the filtrate D after 1-2 hours of dissolution at room temperature and filtration; (6) Ultrafiltration and dialysis Dialyze the filtrate D in step (5) with dissolving solution C at a constant weight of 3-6 times, and concentrate the solution by ultrafiltration to a protein concentration of more than 30 g / L to obtain a stock solution; (7) Preparation Calculate the final volume according to the protein content of the stock solution in step (6) and the specifications of the final product, dilute the stock solution to 23-28 g / L with dialysis buffer, stir the solution, and confirm that the pH value of the product is 6.8-7.2; sterilize the prepared product with a sterilization filter to obtain a semi-finished product; (8) Finished product Package the semi-finished product, freeze-dry the product, seal the product, and perform dry heat inactivation treatment under the conditions of 100±1℃ for 30 minutes to obtain a finished product. The FI precipitate in step (1) is freshly prepared or thawed component I, which is cut into small cubes of about 1 cm in size before dissolution, and then filtered with a 30SP filter to obtain the filtrate A.

2. The process for the preparation of human fibrinogen from Fraction I according to claim 1, characterized in that: The formula of the dissolving solution A is: 29.4 g of sodium citrate, 9.1 g of lysine hydrochloride, and 6000 IU of heparin sodium per kilogram of injection water, and the pH value is 6.8-7.

0.

3. The preparation process for extracting human fibrinogen from component I according to claim 1, characterized in that: The formula of the dissolving solution B is: 15 g of sodium citrate, 9 g of sodium chloride, 3.5 g of lysine hydrochloride, and 10 g of sucrose per kilogram of injection water, and the pH value is 6.8-7.

2.

4. The process for the preparation of human fibrinogen from Fraction I according to claim 1, characterized in that: The formula of the dissolving solution C is: 10 g of sodium citrate, 2 g of glutamic acid sodium, and 20 g of arginine hydrochloride per kilogram of injection water, and the pH value is 6.8-7.

2.

5. The preparation process for extracting human fibrinogen from component I according to claim 1, characterized in that: ​ 6. The process for the preparation of human fibrinogen from Fraction I as claimed in claim 4, wherein the said process is characterized by the steps of: The S / D reagent formulation comprises 11% polysorbate 80 and 3.3% tributyl phosphate.

7. The process for the preparation of human fibrinogen from Fraction I according to claim 1, characterized in that: The specific step of step (2) is: adjusting the filtrate A of step (1) to 6.8-7.2, then adding PEG, stirring and centrifuging, the maximum processing capacity per minute is less than or equal to 2.0 Kg, the centrifugation effluent temperature is controlled at 10-17℃, the dissolved PEG precipitate is filtered by using a 60SP filter core, and the filtrate B is obtained.

8. The process for the preparation of human fibrinogen from Fraction I according to claim 1, characterized in that: The specific step of step (3) is: adjusting the pH of the filtrate B of step (2) to 6.8-7.2, adding S / D reagent at 0.1 times the weight of the product, stirring uniformly, then filtering into an S / D inactivation tank, heating the product to 24-26℃, and performing S / D virus inactivation for at least 6 hours continuously.

9. The process for the preparation of human fibrinogen from Fraction I according to claim 1, characterized in that: The specific step of step (4) is: adjusting the pH of the inactivated liquid of step (3) to 6.8-7.2, adding ethanol after cooling, stirring and centrifuging, the maximum processing capacity per minute is less than or equal to 2.0 Kg, the centrifugation effluent temperature is controlled at -2.5-0℃, the dissolved ethanol precipitate is filtered by using a 60SP filter core, and the filtrate C is obtained.

10. The process for the preparation of human fibrinogen from Fraction I according to claim 1, characterized in that: The specific step of step (5) is: adjusting the pH of the filtrate C of step (4) to 7.1-7.3, then adding glycine, stirring and centrifuging, the maximum processing capacity per minute is less than or equal to 2.0 Kg, the centrifugation effluent temperature is controlled at 0-4℃, the dissolved glycine precipitate is filtered by using a 0.45μm filter core, and the filtrate D is obtained.

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