Preparation method of human immune globulin freeze-dried dosage form for subcutaneous injection

Abstract

The invention relates to a preparation method of a human immune globulin freeze-dried dosage form for subcutaneous injection, and belongs to the technical field of blood products. The preparation method comprises the following steps: (1) performing precipitation separation on components I, II and III in plasma; (2) octanoic acid precipitation separation; (3) primary ultrafiltration; (4) inactivating; (5) anion exchange chromatography; (6) carrying out secondary ultrafiltration; (7) carrying out affinity chromatography; (8) filtering with a nano-membrane; (9) performing ultrafiltration for the third time; and (10) sub-packaging and freeze-drying. Compared with an existing hypodermic injection human immune globulin liquid dosage form, the hypodermic injection human immune globulin freeze-dried dosage form provided by the invention does not need an additional one-way flow ultrafiltration step, and the protein is freeze-dried, can be stored at room temperature and is longer in validity period.

Description

Technical Field

[0001] The present invention relates to the technical field of blood products, and in particular to a method for preparing a freeze-dried dosage form of human immunoglobulin for subcutaneous injection. Background Art

[0002] Subcutaneous injection immunoglobulin (SCIG) is a newer form of immunoglobulin infusion. Compared with intravenous immunoglobulin, subcutaneous immunoglobulin products offer advantages in terms of feasibility for home care treatment and fewer side effects. The US FDA approved ZLB Bering's Vivaglobin and CSL's Hizentra in 2010, respectively. SCIG is readily available and portable, allowing patients to self-administer it at home. It is also suitable for patients with poor intravenous access or those who are responsive to IVIG. Currently approved for the treatment of primary immunodeficiency diseases (PIDs), SCIG offers therapeutic efficacy similar to conventional intravenous administration, making it a safe and effective alternative to intravenous immunoglobulin. SCIG contains approximately 98% pure IgG in a 10% to 20% concentration liquid formulation, reducing the infusion volume. Subcutaneous human immunoglobulin is slowly absorbed by the body after infusion and has relatively stable pharmacokinetics. It is suitable for patients of all ages.

[0003] Currently, the subcutaneous human immunoglobulins that have been marketed or are being registered at home and abroad are all liquid preparations, which generally require additional unidirectional flow ultrafiltration to a concentration of 20%. The process is lengthy and the yield is low. The products are repeatedly ultrafiltered and the protein is repeatedly sheared, which damages the activity of the products to varying degrees. Summary of the Invention

[0004] The present invention addresses the problems of existing subcutaneous human immunoglobulin dosage forms, which are all liquid preparations and easily lose activity during preparation. This method provides a method for preparing a lyophilized formulation of subcutaneous human immunoglobulin to address these issues. This lyophilized formulation eliminates the need for an additional unidirectional flow ultrafiltration step, and the lyophilized protein can be stored at room temperature, extending its shelf life.

[0005] The technical solutions of the present invention are as follows: A method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection comprises the following steps: (1) Separation of plasma components I, II, and III by precipitation; (2) separation by caprylic acid precipitation; (3) primary ultrafiltration; (4) inactivation; (5) anion exchange chromatography; (6) secondary ultrafiltration; (7) affinity chromatography; (8) nanomembrane filtration; (9) tertiary ultrafiltration; (10) packaging and freeze-drying.

[0006] Furthermore, the plasma can be normal plasma, cryoprecipitate-removed plasma, DEAE Sephadex A-50 gel-adsorbed plasma, Capto Heparin-chromatographic plasma, or Capto Q-chromatographic plasma. Necessary measures should be taken before unpacking the plasma to prevent pyrogen and microbial contamination, such as cleaning, disinfection, or other measures. Frozen plasma must first be pulped, with the pulping circulating water temperature ≤37°C.

[0007] Furthermore, in step (1), the preparation of the component I+II+III precipitate includes controlling the plasma pH to 5.9-6.0, adding ethanol to the plasma to control the ethanol concentration in the plasma to 19%, the product temperature to -5.5--4.5°C, and the storage time to 4-15 hours. After the storage is completed, the product is filtered, and diatomaceous earth and perlite are added at a ratio of ≤0.5% by weight of the product and ≤0.5% by weight. During the filtration, the outlet temperature is controlled to be -5.5°C--4.5°C, and the pressure difference is ≤0.24 MPa. The resulting component I+II+III precipitate is stored below -30°C for a storage period of no more than 1 year.

[0008] Furthermore, the step (2) comprises dissolving the component I+II+III precipitate, wherein 5-15 kg of injection water is used to dissolve each 1 kg of the component I+II+III precipitate, the dissolution temperature is 0-26°C, and the dissolution time is 1-3 hours; adjusting the pH value of the product to 4.0-5.0, continuing to dissolve for 1-3 hours, and the dissolution temperature is 0-26°C; then adding octanoic acid / octanoate ion to the solution to 15-50 mmol / kg, adjusting the pH value to 4.0-5.4, continuing to stir for 1-4 hours, and the product temperature is 0-26°C; after stirring, filtering to remove the precipitate, and collecting the supernatant of the filtration. Control the filtration conditions: add perlite at a ratio of 0.5%-5.0% by weight of the product, the pressure difference is ≤0.24 MPa, and the filtrate should be clear and transparent.

[0009] Furthermore, the step (3) is to perform an ultrafiltration treatment on the supernatant obtained in step (2), and the supernatant is concentrated and then dialyzed, and the ultrafiltration membrane has a molecular weight cut-off of ≤50 kD. The ultrafiltration process controls the transmembrane pressure to ≤0.3 MPa.

[0010] Furthermore, the step (4) is to adjust the protein content of the ultrafiltrate to 10-30 g / L, control the pH value to 3.8-4.4, and keep it at 37°C ± 0.5°C for 9 hours to inactivate the virus to obtain an inactivated solution.

[0011] Furthermore, the step (5) is to use Fractogel® EMD DEAE anion exchange chromatography and Fractogel® EMD TMAE anion exchange chromatography respectively; the column filler used in the Fractogel® EMD DEAE anion exchange chromatography is Fractogel® EMD After low-pH incubation, adjust the pH of the product to 4.0-5.4 and the conductivity to 0.5-1.5 mS / cm. Filter the inactivation solution and the solution through a filter cartridge with a pore size of ≤0.5 μm before chromatography. First, treat the column with 3-6 column volumes of pre-equilibration solution (200-400 mmol / kg acetic acid-sodium acetate solution, pH 4.0-5.4). Then, treat the column with 3-6 column volumes of equilibration solution (10-25 mmol / kg acetic acid-sodium acetate solution, pH 4.0-5.4). The adjusted inactivation solution is passed through the equilibrated column, allowing human immunoglobulin to flow through, while contaminants are adsorbed on the column. Wash with the equilibration solution to displace proteins from the column, and collect the flow-through product. The flow rate for pre-equilibration, equilibration, sample loading, and wash lines should be 40-120 cm / h.

[0012] Column cleaning: Use eluent, 0.5 mol / kg sodium hydroxide, and water for injection. Eluent: 100-400 mmol / kg acetic acid-sodium acetate solution, sodium chloride content ≥ 800 mmol / kg, pH 4.0-5.4.

[0013] Column storage: 20% ethanol or 0.01 mol / kg sodium hydroxide.

[0014] The Fractogel® EMD TMAE anion exchange chromatography column is packed with Fractogel® EMD TMAE. The pH of the product after Fractogel® EMD DEAE anion exchange chromatography was adjusted to 4.6-6.0, and the conductivity to 0.8-1.8 mS / cm. Both the product and the solution were filtered using a filter cartridge with a pore size of 0.5 μm or less before chromatography. The column was first treated with a pre-equilibration solution (200-400 mmol / kg acetic acid-sodium acetate solution, pH 4.6-6.0) for 3-6 column volumes. The column was then treated with an equilibration solution (10-25 mmol / kg acetic acid-sodium acetate solution, pH 4.6-6.0) for 3-6 column volumes. The adjusted product was passed through the equilibrated column, with human immunoglobulin flowing through and contaminants being adsorbed on the column. The column was then washed with the equilibration solution to displace proteins, and the product that flowed through was collected. The flow rate of the pre-equilibration, equilibration, loading and washing lines is 40~120 cm / h.

[0015] Column cleaning: Use eluent, 0.5 mol / kg sodium hydroxide, and water for injection. Eluent: 100-400 mmol / kg acetic acid-sodium acetate solution with a sodium chloride content ≥800 mmol / kg and a pH of 4.6-6.0.

[0016] Column storage: 20% ethanol or 0.01 mol / kg sodium hydroxide.

[0017] Furthermore, the step (6) is to perform a second ultrafiltration on the product after Fractogel® EMD TMAE chromatography, and then perform dialysis after the product is concentrated, and the ultrafiltration membrane molecular weight cutoff is ≤50 kD; the transmembrane pressure during the ultrafiltration process is controlled to be ≤0.3 MPa.

[0018] Furthermore, the step (7) is to perform affinity chromatography on the solution of the secondary ultrafiltration, wherein the column fillers used in the affinity chromatography are ESHMUNO® P anti-A gel and ESHMUNO® P anti-B gel, and the ratio of the volume of ESHMUNO® P anti-A gel to the volume of ESHMUNO® P anti-B gel is in the range of 1 to 3:1; the solution of the secondary ultrafiltration and the solution before chromatography are both filtered using a filter element with a pore size of ≤0.5 μm; first, the chromatography column is treated with a pre-equilibration solution for 3 to 6 column volumes; the pre-equilibration solution is 200 to 500 mmol / kg acetic acid-sodium acetate solution, pH value 4.0 to 5.0; then, the chromatography column is treated with an equilibrium solution for 3 to 6 column volumes, wherein the equilibrium solution is 0.5 to 10 mmol / kg acetic acid-sodium acetate solution, pH value 4.0 to 5.0; the adjusted product passes through the equilibrated chromatography column, the human immunoglobulin flows through the chromatography column, and the impurity protein is adsorbed on the chromatography column. Use the equilibration solution to displace the protein from the column and collect the flow-through. The flow rate for pre-equilibration, equilibration, loading, and washing should be 40-120 cm / h.

[0019] Column cleaning: Use eluent, 0.5 mol / kg sodium hydroxide, cleaning solution, and water for injection. Eluent: 80-120 mmol / kg glycine solution, pH 2.6-2.8. Cleaning solution: 100-500 mmol / kg acetic acid-sodium acetate solution, sodium chloride content ≥ 800 mmol / kg, pH 4.0-5.0.

[0020] Column storage: 20% ethanol or 0.01 mol / kg sodium hydroxide.

[0021] Furthermore, step (8) is as follows: first, add 10-30 g / kg of glycine to the solution obtained by affinity chromatography, control the pH value to 3.5-4.5, the protein content in the solution obtained by affinity chromatography is ≤ 60 g / L, pre-filter with a 0.1 μm filter membrane, and filter with a nanomembrane filter to remove viruses. The transmembrane pressure difference, filtration time, and filtration load control should meet the requirements of the nanomembrane filter and verification. The nanomembrane should be integrity tested before and after use.

[0022] Furthermore, step (9) is as follows: the solution after nano-membrane filtration is subjected to three ultrafiltrations, and the ultrafiltration dialysate is dialyzed, and the ultrafiltration membrane has a molecular weight cut-off of ≤50 kD. The ultrafiltration dialysate is a 10-30 g / kg glycine solution with a pH of 3.5-4.5. The transmembrane pressure during the ultrafiltration process is controlled to be ≤0.3 MPa. The product after ultrafiltration is the stock solution.

[0023] Furthermore, the step (10) is as follows: the protein content is ≥100 g / L, and the auxiliary materials are: glycine at 10-30 g / kg; polysorbate 80 at 20-80 μg / ml. The pH value is controlled at 4.0-5.0; the specification is 4g / bottle.

[0024] The freeze-drying process is as follows: after entering the cabinet, the temperature is lowered to -10~0℃ and kept constant for 2~4 hours; the temperature is lowered to -55~-45℃ and kept constant at -55~-45℃ for 2~6 hours; the temperature begins to rise to -20~-10℃ and kept constant at -20~-10℃ for 1~4 hours; the temperature is lowered to -55~-45℃ and kept constant at -55~-45℃ for 4~10 hours, and vacuum sublimation drying is started; within 4~10 hours, the temperature is raised to -15~-5℃ and kept constant at -15~-5℃ for 60~120 hours; within 2~10 hours, the temperature is raised to -5~5℃ and kept constant at -5~5℃ for 2~10 hours; within 2~10 hours, the temperature is raised to 28~35℃ and kept constant at 28~35℃ for 6~12 hours.

[0025] Stoppering and unloading: The stopper is pressed under vacuum conditions, and the product is capped after unloading. This should comply with the "Biological Product Subpackaging and Storage and Transportation Management" regulations of the 2020 edition of the "Chinese Pharmacopoeia".

[0026] Sterilization: It should comply with the "Biological Products Subpackaging, Storage and Transportation Management" regulations of the 2020 edition of the "Chinese Pharmacopoeia".

[0027] Repackaging: It should comply with the relevant provisions of the "Repackaging and Storage and Transportation Management of Biological Products" and General Chapter 0102 of the 2020 edition of the "Chinese Pharmacopoeia".

[0028] The vacuum degree and appearance of the subcutaneous human immunoglobulin lyophilized dosage form prepared by the present invention shall be inspected and shall comply with the provisions of the "Biological Product Subpackaging, Storage and Transportation Management" of the 2020 edition of the "Chinese Pharmacopoeia".

[0029] The packaging of lyophilized human immunoglobulin for subcutaneous injection should comply with the "Biological Products Subpackaging, Storage and Transportation Management" and General Chapter 0102 of the 2020 edition of the Chinese Pharmacopoeia. Packaging: 1 bottle / box, with an additional 20 ml bottle of sterile water for injection for reconstitution.

[0030] When the subcutaneous human immunoglobulin lyophilized preparation prepared by the present invention is used, 20 ml of sterile injection water needs to be added to each bottle of lyophilized preparation for reconstitution.

[0031] The beneficial effects of the present invention are: Compared with the existing liquid dosage form of human immunoglobulin for subcutaneous injection, the freeze-dried dosage form of human immunoglobulin for subcutaneous injection provided by the present invention does not require an additional unidirectional flow ultrafiltration step, and the protein can be stored at room temperature after freeze-drying and has a longer shelf life. DETAILED DESCRIPTION

[0032] In order to enable those skilled in the art to better understand the technical solutions of the present invention, the technical solutions in the embodiments of the present invention are clearly and completely described below. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work should fall within the scope of protection of the present invention.

[0033] Example 1 A method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection comprises the following steps: (1) Separation of components I+II+III by precipitation Before opening the plasma bag, cleaning and disinfection measures should be taken to prevent pyrogen and microbial contamination. The raw material used is frozen plasma, a total of 750 kg, and the pulping temperature is: the pulping circulating water temperature is ≤37°C, and the plasma temperature is ≤10°C.

[0034] The pH of the plasma obtained by slurrying was controlled at 5.9-6.0. Ethanol was added to maintain the ethanol concentration at 19%. The product was kept at a temperature of -5.5-4.5°C and allowed to stand for 12 hours. After standing, the product was filtered, and 0.5 kg of diatomaceous earth and 0.5 kg of perlite were added as filter aids. During the filtration, the outlet temperature was controlled at -5.5-4.5°C, and the pressure differential was ≤0.24 MPa. A total of 70 kg of precipitate containing components I, II, and III was obtained and stored below -30°C.

[0035] (2) Caprylic acid precipitation and separation The precipitate of components I+II+III obtained in step (1) was dissolved in 700 kg of water for injection at a temperature of 6°C for 1 hour. The pH value of the product was adjusted to 4.5, and the solution was dissolved for 1.5 hours at a temperature of 6°C. Caprylic acid / caprylate ion was then added to the solution to a value of 30 mmol / kg, the pH value was adjusted to 5.0, and stirring was continued for 3 hours at a temperature of 10°C. After stirring, the precipitate was removed by filtration, and the supernatant was collected. 25 kg of perlite was added during the filtration, and the pressure difference was ≤0.24 MPa. The filtrate was clear and transparent.

[0036] (3) Primary ultrafiltration The supernatant obtained in step (2) is subjected to a first ultrafiltration treatment. The supernatant is concentrated and then dialyzed. The molecular weight cut-off of the ultrafiltration membrane is ≤50 kD. The transmembrane pressure during the ultrafiltration process is controlled to be ≤0.3 MPa.

[0037] (4) Inactivation The protein content of the primary ultrafiltrate was adjusted to 30 g / L, the pH value was controlled at 4.2, and the virus was inactivated by keeping the mixture at 37°C ± 0.5°C for 9 hours to obtain an inactivated solution.

[0038] (5) Anion exchange chromatography (a) Fractogel® EMD DEAE anion exchange chromatography The column was packed with Fractogel® EMD DEAE. After low-pH incubation, the pH of the product was adjusted to 4.8 and the conductivity to 1.0 mS / cm. Both the inactivation solution and the solution were filtered through a filter cartridge with a pore size of ≤0.5 μm before chromatography. The column was first treated with 4 column volumes of pre-equilibration solution (300 mmol / kg acetic acid-sodium acetate solution, pH 4.8). The column was then treated with 4 column volumes of equilibration solution (20 mmol / kg acetic acid-sodium acetate solution, pH 4.8). The adjusted inactivation solution was passed through the equilibrated column, allowing human immunoglobulin to flow through, while contaminants were adsorbed on the column. The column was washed with equilibration solution to displace proteins, and the flow-through product was collected. The flow rate during pre-equilibration, equilibration, sample loading, and washes was 80 cm / h.

[0039] (b) Fractogel® EMD TMAE anion exchange chromatography The column was packed with Fractogel® EMD TMAE. After anion exchange chromatography with Fractogel® EMD DEAE, the pH of the product was adjusted to 5.6 and the conductivity to 1.4 mS / cm. Both the product and the solution were filtered using a filter cartridge with a pore size of 0.5 μm or less before chromatography. The column was first treated with 4 column volumes of pre-equilibration solution (300 mmol / kg acetic acid-sodium acetate solution, pH 5.6) and then with 4 column volumes of equilibration solution (20 mmol / kg acetic acid-sodium acetate solution, pH 5.6). The adjusted product was passed through the equilibrated column, and human immunoglobulin flowed through the column, while contaminants were adsorbed on the column. The column was washed with equilibration solution to displace proteins, and the flow-through product was collected. The flow rate for pre-equilibration, equilibration, sample loading, and wash lines was 80 cm / h.

[0040] (6) Secondary ultrafiltration The Fractogel® EMD TMAE chromatography product was subjected to a second ultrafiltration step. The product was concentrated and then dialyzed. The ultrafiltration membrane had a molecular weight cutoff of ≤50 kD. The transmembrane pressure during the ultrafiltration process was controlled to ≤0.3 MPa.

[0041] (7) Affinity chromatography The secondary ultrafiltration solution was subjected to affinity chromatography using ESHMUNO® Panti-A gel and ESHMUNO® P anti-B gel as column packings, with a volume ratio of ESHMUNO® P anti-A gel to ESHMUNO® Panti-B gel of 1.5:1. Both the secondary ultrafiltration solution and the solution prior to chromatography were filtered through a filter cartridge with a pore size of 0.5 μm or less. The column was first treated with a pre-equilibration solution (350 mmol / kg acetic acid-sodium acetate solution, pH 4.5) for four column volumes, followed by a equilibration solution (5 mmol / kg acetic acid-sodium acetate solution, pH 4.5) for four column volumes. The adjusted product was then passed through the equilibrated column, with human immunoglobulin flowing through and contaminants being adsorbed on the column. The column was then washed with the equilibration solution to displace proteins, and the flow-through product was collected. The flow rate for pre-equilibration, equilibration, sample loading, and washes was 80 cm / h.

[0042] (8) Nanomembrane filtration First, add 20 g / kg of glycine to the solution obtained by affinity chromatography to control the pH value to 4.0. The protein content in the solution obtained by affinity chromatography is ≤ 60 g / L. Pre-filter with a 0.1 μm filter membrane and filter with a nanomembrane filter to remove viruses.

[0043] (9) Three-stage ultrafiltration The solution after nano-membrane filtration was subjected to three ultrafiltration cycles, and the dialysate was dialyzed using an ultrafiltration membrane with a molecular weight cut-off of ≤50 kD. The dialysate consisted of a 20 g / kg glycine solution with a pH of 4.0. The transmembrane pressure was controlled to ≤0.3 MPa during the ultrafiltration process. The product after ultrafiltration was the stock solution.

[0044] (10) Packaging and freeze-drying Prepare the product to a protein content of ≥100 g / L. Excipients: glycine at 20 g / kg and polysorbate 80 at 50 μg / ml. Maintain a pH of 4.5. 4 g per bottle.

[0045] The freeze-drying process is as follows: after entering the cabinet, the temperature is reduced to -4℃ and kept constant for 2 hours; the temperature is reduced to -50℃ and kept constant at 050℃ for 5 hours; the temperature begins to return to -20~-10℃-15, and the temperature is kept constant at -15℃ for 2 hours; the temperature is reduced to -50℃, and the temperature is kept constant at -50℃ for 6 hours, and vacuum sublimation drying is started; within 6 hours, the temperature is increased to -10℃, and the temperature is kept constant at -10℃ for 100 hours; within 8 hours, the temperature is increased to 0℃, and the temperature is kept constant at 0℃ for 5 hours; within 4 hours, the temperature is increased to 32℃, and the temperature is kept constant at 32℃ for 8 hours.

[0046] Stoppering and unloading: The stopper is pressed under vacuum conditions, and the product is capped after unloading. This should comply with the "Biological Product Subpackaging and Storage and Transportation Management" regulations of the 2020 edition of the "Chinese Pharmacopoeia".

[0047] Sterilization: It should comply with the "Biological Products Subpackaging, Storage and Transportation Management" regulations of the 2020 edition of the "Chinese Pharmacopoeia".

[0048] Repackaging: It should comply with the relevant provisions of the "Repackaging and Storage and Transportation Management of Biological Products" and General Chapter 0102 of the 2020 edition of the "Chinese Pharmacopoeia".

[0049] The vacuum degree and appearance of the subcutaneous human immunoglobulin lyophilized dosage form prepared by the present invention shall be inspected and shall comply with the provisions of the "Biological Product Subpackaging, Storage and Transportation Management" of the 2020 edition of the "Chinese Pharmacopoeia".

[0050] The packaging of lyophilized human immunoglobulin for subcutaneous injection should comply with the "Biological Products Subpackaging, Storage and Transportation Management" and General Chapter 0102 of the 2020 edition of the Chinese Pharmacopoeia. Packaging: 1 bottle / box, with an additional 20 ml bottle of sterile water for injection for reconstitution.

[0051] Test Case The performance of the subcutaneous human immunoglobulin lyophilized preparation prepared in Example 1 was compared with the existing subcutaneous human immunoglobulin liquid injection. The subcutaneous human immunoglobulin liquid injection was selected from a batch of samples produced by our company. The detection method is based on the 2020 edition of the Pharmacopoeia, and is as follows: Table 1 - Stability test results

[0052] Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Without departing from the spirit and substance of the present invention, persons of ordinary skill in the art may make various equivalent modifications or substitutions to the embodiments of the present invention, and such modifications or substitutions shall be within the scope of the present invention. Any changes or substitutions that can be readily conceived by persons skilled in the art within the technical scope disclosed in the present invention shall be within the scope of protection of the present invention.

Claims

1. A method for preparing a freeze-dried dosage form of human immunoglobulin for subcutaneous injection, characterized in that: The following steps are involved: (1) Separation of plasma components I, II, and III by precipitation; (2) separation by caprylic acid precipitation; (3) primary ultrafiltration; (4) inactivation; (5) anion exchange chromatography; (6) secondary ultrafiltration; (7) affinity chromatography; (8) nanomembrane filtration; (9) tertiary ultrafiltration; (10) packaging and freeze-drying.

2. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 1, wherein: The plasma is ordinary plasma, plasma removed from cryoprecipitate, plasma adsorbed by DEAE Sephadex A-50 gel, plasma chromatographed by Capto Heparin or plasma chromatographed by Capto Q. Frozen plasma needs to be pulped first, wherein the pulping circulating water temperature is ≤37°C.

3. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 1, wherein: The step (1) is as follows: controlling the plasma pH to 5.9-6.0, adding ethanol to the plasma, controlling the ethanol concentration in the plasma to 19%, the product temperature to -5.5--4.5°C, and the standing time to 4-15 hours; after the standing time is completed, filtering, and controlling the outlet temperature to -5.5--4.5°C during the filtering, and the pressure difference to ≤0.24 MPa.

4. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 3, wherein: The step (2) comprises dissolving the component I+II+III precipitate, wherein 5-15 kg of injection water is used to dissolve each 1 kg of the component I+II+III precipitate, the dissolution temperature is 0-26°C, and the dissolution time is 1-3 hours; adjusting the pH value of the product to 4.0-5.0, continuing to dissolve for 1-3 hours, and the dissolution temperature is 0-26°C; then adding octanoic acid / octanoate ion to the solution to 15-50 mmol / kg, adjusting the pH value to 4.0-5.4, continuing to stir for 1-4 hours, and the product temperature is 0-26°C; after the stirring is completed, filtering to remove the precipitate, and collecting the filter press supernatant.

5. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 4, wherein: The step (3) is to perform an ultrafiltration treatment on the supernatant obtained in the step (2), and then dialyze the supernatant after concentration, with the ultrafiltration membrane having a molecular weight cut-off of ≤50 kD; and the transmembrane pressure is controlled to be ≤0.3 MPa during the ultrafiltration process.

6. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 5, wherein: The step (4) is to adjust the protein content of the ultrafiltrate to 10-30 g / L, control the pH value to 3.8-4.4, and keep it at 37°C ± 0.5°C for 9 hours to inactivate the virus and obtain an inactivated solution.

7. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 6, wherein: The step (5) is to use Fractogel® EMD DEAE anion exchange chromatography and Fractogel® EMDTMAE anion exchange chromatography respectively; the column filler used in the Fractogel® EMD DEAE anion exchange chromatography is Fractogel® EMD DEAE, and the pH value of the product after low pH incubation is adjusted to 4.0-5.4, and the conductivity is adjusted to 0.5-1.5 mS / cm; the inactivation solution and the solution are filtered with a filter element with a pore size of ≤0.5 μm before chromatography; first, the chromatography column is treated with a pre-equilibration solution for 3-6 column volumes, the pre-equilibration solution is 200-400 mmol / kg acetic acid-sodium acetate solution, pH value 4.0-5.4; then, the chromatography column is treated with an equilibrium solution for 3-6 column volumes, the equilibrium solution is 10-25 mmol / kg acetic acid-sodium acetate solution, pH value 4.0-5.4; the flow rate of the pre-equilibration solution and the equilibrium solution is 40-120 cm / h; The Fractogel® EMD TMAE anion exchange chromatography column filler is Fractogel® EMD TMAE; the pH value of the product after Fractogel® EMD DEAE anion exchange chromatography is adjusted to 4.6-6.0, and the conductivity is adjusted to 0.8-1.8 mS / cm; the product and solution are filtered using a filter element with a pore size of ≤0.5 μm before chromatography; the chromatography column is first treated with a pre-equilibration solution for 3-6 column volumes, wherein the pre-equilibration solution is a 200-400 mmol / kg acetic acid-sodium acetate solution with a pH value of 4.6-6.0; and then the chromatography column is treated with an equilibration solution for 3-6 column volumes, wherein the equilibration solution is a 10-25 mmol / kg acetic acid-sodium acetate solution with a pH value of 4.6-6.0; the linear flow rate of the pre-equilibration solution and the equilibration solution is 40-120 cm / h.

8. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 7, wherein: The step (6) is to perform a second ultrafiltration on the product after Fractogel® EMD TMAE chromatography, and then perform dialysis after the product is concentrated, with the ultrafiltration membrane having a molecular weight cutoff of ≤50 kD; and the transmembrane pressure is controlled to be ≤0.3 MPa during the ultrafiltration process.

9. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 8, wherein: The step (7) is to perform affinity chromatography on the solution of the secondary ultrafiltration, wherein the column fillers used in the affinity chromatography are ESHMUNO® P anti-A gel and ESHMUNO® P anti-B gel, and the ratio of the volume of ESHMUNO® P anti-A gel to the volume of ESHMUNO® P anti-B gel is in the range of 1 to 3:1; the solution of the secondary ultrafiltration and the solution are filtered using a filter element with a pore size of ≤0.5 μm before chromatography; first, the chromatography column is treated with a pre-equilibration solution for 3 to 6 column volumes; the pre-equilibration solution is 200 to 500 mmol / kg acetic acid-sodium acetate solution with a pH value of 4.0 to 5.0; and then the chromatography column is treated with an equilibrium solution for 3 to 6 column volumes, wherein the equilibrium solution is 0.5 to 10 mmol / kg acetic acid-sodium acetate solution with a pH value of 4.0 to 5.0; the linear flow rate of the pre-equilibration solution and the equilibrium solution is 40 to 120 cm / h.

10. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 9, wherein: The step (8) is as follows: first, 10-30 g / kg of glycine is added to the solution obtained by affinity chromatography, the pH value is controlled to be 3.5-4.5, the protein content in the solution obtained by affinity chromatography is ≤60 g / L, pre-filtering is performed with a 0.1 μm filter membrane, and filtering is performed with a nano-membrane filter.

11. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 10, characterized in that: The step (9) is as follows: the solution after nano-membrane filtration is ultrafiltered three times, and the ultrafiltration dialysate is dialyzed, and the ultrafiltration membrane molecular weight cut-off is ≤50kD; the ultrafiltration dialysate is 10-30g / kg glycine solution, and the pH value is 3.5-4.5; the ultrafiltration process controls the transmembrane pressure to ≤0.3MPa.

12. The method for preparing a lyophilized dosage form of human immunoglobulin for subcutaneous injection according to claim 11, wherein: The step (10) is as follows: the preparation is carried out according to the protein content ≥100 g / L, and the auxiliary materials are: glycine is 10~30 g / kg; polysorbate 80 is 20~80 μg / ml; the pH value is controlled at 4.0~5.0; the specification is 4g / bottle; the freeze-drying process is as follows: after the cabinet is put into operation, the temperature is reduced to -10~0°C and kept constant for 2~4 hours; the temperature is reduced to -55~-45°C and kept constant at -55~-45°C for 2~6 hours; the temperature is started to return to -20~-10°C and kept constant at -20~-10°C for 1~4 hours; the temperature is reduced to -55~-45°C and kept constant at -55~-45°C for 4~10 hours, and vacuum sublimation drying is started; within 4~10 hours, the temperature is increased to -15~-5°C and kept constant at -15~-5 ℃ and keep constant temperature for 60~120 hours; within 2~10 hours, increase the temperature to -5~5 ℃ and keep constant temperature at -5~5 ℃ for 2~10 hours; within 2~10 hours, increase the temperature to 28~35 ℃ and keep constant temperature at 28~35 ℃ for 6~12 hours.

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