CN112521487A - Improved production process of human serum albumin - Google Patents
Improved production process of human serum albumin Download PDFInfo
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- CN112521487A CN112521487A CN202011535109.7A CN202011535109A CN112521487A CN 112521487 A CN112521487 A CN 112521487A CN 202011535109 A CN202011535109 A CN 202011535109A CN 112521487 A CN112521487 A CN 112521487A
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- 108091006905 Human Serum Albumin Proteins 0.000 title claims abstract description 20
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- 101000623895 Bos taurus Mucin-15 Proteins 0.000 claims abstract description 65
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- 230000008018 melting Effects 0.000 claims abstract description 11
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Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/76—Albumins
- C07K14/765—Serum albumin, e.g. HSA
Abstract
The invention relates to an improved human serum albumin production process, which comprises plasma melting, detection, component I preparation, component II + III preparation, component IV preparation, component V refining, ultrafiltration, diluted preparation, pasteurization, split charging and product incubation, wherein whether the corresponding steps meet the production requirements or not is judged according to precipitates obtained by pressure filtration in the steps of the component I preparation, the component II + III preparation, the component IV preparation and the component V preparation. The invention firstly detects the components and the content of the protein in the raw material plasma, and in the subsequent production, the weight of the filter-pressing sediment of each component is weighed to judge whether the component meets the requirement of purification, thereby reducing the detection procedures in the production and improving the production efficiency.
Description
Technical Field
The invention relates to a separation and purification process of protein drugs in the field of biological pharmacy, in particular to an improved production process of human serum albumin.
Background
Human serum albumin is the main product of blood preparations and is also the final product of the backbone of the plasma protein production process. Albumin is synthesized in liver cells, has the characteristics of extremely high synthesis speed, relatively small molecules (molecular weight of 66250), relatively large surface area, relatively symmetrical molecular configuration, simple and firm structure, high toughness and the like. Albumin has important function for hemorrhagic shock, serious burn, encephaledema, and blood disease of liver and kidney diseases. Since Cohn et al established the technology of low temperature ethanol human plasma protein isolation in the 40 s, the industrial mass production of plasma protein has advanced greatly. In the production of human serum albumin, other impurity proteins in the plasma are gradually separated out and separated by adjusting the concentration of ethanol at different stages, and finally the required protein components in the plasma are reserved. In the existing production process, because the components and the content of the blood plasma as a raw material are relatively stable, the amount of low-temperature ethanol added in each process is also stable, and the low-temperature ethanol does not need to be adjusted too much, and the low-temperature ethanol can be divided into the following types according to the components of protein in a solution according to the sequence of production processes: the component I, the component II + III, the component IV and the component V are all provided with impurity proteins, so that low-temperature ethanol is needed to be used for gradual purification, the purification effect of the former component can directly influence the treatment of the latter component, but in the prior art, whether the purification of the former component meets the production requirement of the latter component can be determined only through multiple detections, and the detection procedures are more, so that the production efficiency is influenced.
Disclosure of Invention
Aiming at the defects in the prior art, the invention aims to provide an improved human serum albumin production process, which comprises the steps of firstly detecting the components and the content of protein in raw plasma, and weighing the weight of filter-pressing precipitate of each component in subsequent production to judge whether the component meets the requirement of purification, thereby reducing the detection procedures in production and improving the production efficiency.
The technical purpose of the invention is realized by the following technical scheme:
an improved human serum albumin production process comprises plasma melting, detection, component I preparation, component II + III preparation, component IV preparation, component V refining, ultrafiltration, diluted blending, pasteurization, protein sterilization and split charging and product incubation, wherein in the steps of the component I preparation, the component II + III preparation, the component IV preparation and the component V preparation, whether the corresponding steps meet the production requirements is judged according to the sediment obtained by filter pressing, the specific production steps are as follows,
slurry melting: the method comprises the following steps of (1) breaking a bag of raw plasma, conveying the raw plasma to a plasma melting tank, performing interlayer circulation plasma melting by using water at the temperature of 30-35 ℃, stopping water circulation after the plasma is melted, controlling the temperature of the plasma to be 0-4 ℃, performing centrifugal separation and precipitation, and conveying the plasma after the precipitation is removed to a first reaction tank for extraction and separation;
and (3) detection: sampling from the reaction tank for detection, and respectively detecting the mass M of the substances to be separated in the steps of preparing the component I, preparing the components II and III, preparing the component IV and preparing the component V1、M2、M3And M4；
Preparing a component I: after plasma enters a first reaction tank, stirring is started, the temperature of the plasma is controlled to be 0-1 ℃, a buffer solution with the pH value of 4.0 is dripped to adjust the pH value to be 6.80-7.00, low-temperature ethanol is added until the volume specific concentration of the ethanol is 8%, the temperature is controlled to be-1-3 ℃ after the ethanol is added, centrifugation is carried out after the ethanol is added, a component I precipitate and a component I supernatant are obtained, and the mass of the component I precipitate is weighed to be m1Comparison of the component I precipitation masses m1Mass M of substance to be separated from component I1If the mass m of the component I precipitate1Mass M of substance to be separated greater than component I1Then the supernatant of the component I enters the next working procedure, if the mass m of the precipitation of the component I1Less than or equal to component IMass M of the dissociated substance1Adjusting the production parameters of the first reaction tank, adjusting the input amount of low-temperature ethanol to increase the mass of the component I precipitate, performing filter pressing again, and comparing the total mass m of the component I precipitate1Mass M of substance to be separated from component I1Repeating the operation until the component I precipitate is completely separated out, and allowing the component I supernatant to enter the next step, wherein the component I precipitate is used for preparing human fibrinogen;
preparing a component II + III: taking the supernatant of the component I into a second reaction tank, dropwise adding a buffer solution with the pH of 4.0 to adjust the pH value of the supernatant of the component I to be 6.80-6.85, adding low-temperature ethanol until the volume concentration of the ethanol is 20%, controlling the temperature to be-4-6 ℃ after adding the ethanol, controlling the pH to be 6.80-7.00 after adding the ethanol, stirring, standing, starting stirring, then performing filter pressing, and obtaining a component II + III precipitate and a component II + III supernatant after filter pressing, wherein the mass of the component II + III precipitate is m2Comparison of the component II + III precipitation masses m2Mass M of substance to be separated from Components II + III2If the mass m of the component II + III precipitates2Mass M of substance to be separated greater than components II + III2Then the supernatant of the component II + III enters the next working procedure, if the mass m of the component II + III precipitate is2Mass M less than or equal to mass of substance to be separated of Components II + III2Adjusting the production parameters of the second reaction tank, adjusting the input amount of low-temperature ethanol to increase the mass of the component II + III precipitate, performing filter pressing again, and comparing the total mass m of the component II + III precipitate2Mass M of substance to be separated from Components II + III2Repeating the operation until the component II + III precipitates are completely separated out, and allowing the component II + III supernatant to enter the next step, wherein the component II + III precipitates are used for preparing immunoglobulin;
preparing a component IV: taking the component II + III supernatant fluid into a third reaction tank, dropwise adding a buffer solution with the pH of 4.0 to adjust the pH value of the component II + III supernatant fluid to be 5.70-5.90, controlling the temperature in the third reaction tank to be-4-6 ℃, adding low-temperature ethanol until the volume ratio concentration of the ethanol is 40%, after the ethanol is added, the pH value of the liquid is 5.90-6.10, stirring, standing, then starting stirring, then performing pressure filtration, and obtaining the finished product after pressure filtrationComponent IV precipitate and component IV supernatant, weighing the component IV precipitate to m3Comparison of the precipitated masses m of component IV3Mass M of substance to be separated from component IV3If component IV precipitates by mass m3Mass M of substance to be separated greater than component IV3The supernatant of the component IV enters the next working procedure, and if the component IV precipitates by mass m3Mass M less than or equal to that of the substance to be separated of component IV3Adjusting the production parameters of the third reaction tank, adjusting the input amount of low-temperature ethanol to increase the mass of the component I precipitate, performing filter pressing again, and comparing the total mass m of the component IV precipitate3Mass M of substance to be separated from component IV3Repeating the operation until the component IV precipitate is completely separated out, and allowing the component IV supernatant to enter the next step, wherein the component IV precipitate is discarded;
preparing a component V: taking the supernatant of the component IV, adding a buffer solution with the pH of 4.0 dropwise into a fourth reaction tank to adjust the pH value of the supernatant of the component IV to be 4.80-5.00, controlling the temperature in the fourth reaction tank to be below-8 ℃, stirring, performing filter pressing, and obtaining a component V precipitate and a component V supernatant after filter pressing, wherein the mass of the component V precipitate is weighed to be m4Comparison of the Mass m of the component V precipitate4Mass M of substance to be separated from component V4If component V precipitates a mass m4Mass M of the substance to be separated greater than component V4Then the component V is precipitated and enters the next working procedure, if the component V is precipitated by mass m4Mass M of the substance to be separated of fraction V or less4Adjusting the production parameters of the fourth reaction tank to increase the mass of the component V precipitate, and comparing the total mass m of the component V precipitate after filter pressing again4Mass M of substance to be separated from component V4Repeating the operation until the component V precipitate is completely separated out, and entering the next step, wherein the component V supernatant is transferred to an alcohol recovery tower to recover alcohol;
refining a component V: taking the component V precipitate, fully dissolving the component V precipitate with 5 times of injection water, dropwise adding a buffer solution with the pH of 4.0 to adjust the pH value of a component V precipitate dissolving solution to be 4.50-4.60, adding low-temperature ethanol until the volume ratio concentration of the ethanol is 10%, controlling the temperature to be-2-3 ℃ after adding the ethanol, stirring, and performing filter pressing to obtain a component V purified solution;
and (3) ultrafiltration: controlling the feed hydraulic pressure to be less than or equal to 0.45Mpa, the reflux pressure to be less than or equal to 0.30Mpa and the pressure difference between the feed hydraulic pressure and the reflux pressure to be greater than 0.10Mpa in the ultrafiltration process, concentrating the component V purified solution until the protein content is more than 15%, performing equal-volume dialysis on the concentrated component V purified solution by sequentially using 4 times of volume of 0.85% sodium chloride solution at 2-8 ℃ and 5 times of volume of 0.5% sodium chloride solution at 2-8 ℃, concentrating the protein concentration to be more than 20% after the dialysis is finished, and adjusting the pH value of the protein solution to be 6.90-7.10 by using 1mol/L NaHCO 3;
diluting and preparing: diluting according to 20% protein concentration, respectively adding sodium caprylate as protective agent to 0.16mmol/g protein concentration and sodium chloride to 145mmol/L final stock solution, adding water for injection to the final diluted amount, and stirring well.
And (3) pasteurizing inactivation: after the diluted preparation solution is uniformly stirred, jacket circulating water at 60.5 ℃ is used for heating, when the temperature of the protein solution rises to 59.5 ℃, timing is started, inactivation is carried out at 59.5-60.5 ℃ for 12 hours, and after the inactivation is finished, the temperature of the protein solution is reduced to 20-30 ℃.
Protein sterilization, subpackaging and product incubation: and (3) sterilizing and filtering the inactivated protein liquid by using a 0.2-micron sterilizing filter element, filling according to requirements, transferring the filled product to a hatching room at the temperature of 30-32 ℃, incubating for 14 days, and performing finished product verification after incubation.
In one embodiment, the fraction I precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass of the solid in the fraction I precipitate after ethanol removal is n1The calculation method is as follows,
taking the mass as a after filter pressing1The precipitate of component I is evaporated to dryness at low temperature to obtain the product with mass b1The mass percentage content of the solid in the component I precipitate is calculated as x1Wherein, in the step (A),
then according to the mass percent content x1To calculate the mass n of the solid in the component I precipitate after removing ethanol1I.e. n1＝m1×x1；
When n is1＝M1When n is less than n, the impurity substance in the component I is completely precipitated1＜M1In the meantime, it is indicated that the impurity substances in the component I are not completely precipitated and need to be continuously separated.
In one embodiment, the component II + III precipitate still contains a small amount of ethanol solution after filter pressing, wherein the mass n of the solid in the component II + III precipitate after ethanol removal is n2The calculation method is as follows,
taking the mass as a after filter pressing2The precipitate of the components II and III is evaporated to dryness at low temperature to obtain the product with the mass b2The mass percentage of the solid in the component II + III precipitate is calculated as x2Wherein, in the step (A),
then according to the mass percent content x2To calculate the mass n of the solid in the component II + III precipitate after removing ethanol2I.e. n2＝m2×x2；
When n is2＝M2When n is less than n, the impurity substances in the components II + III are completely separated out2＜M2In the meantime, it is indicated that the impurity substances in the components II + III are not completely separated out and need to be continuously separated.
In one embodiment, the fraction IV precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass of the solid in the fraction IV precipitate after ethanol removal is n3The calculation method is as follows,
taking the mass as a after filter pressing3The precipitate of component IV is evaporated to dryness at low temperature to obtain the product with mass b3The mass percentage content of the solid in the component IV precipitate is calculated as x3Wherein, in the step (A),
then according to the mass percent content x3To calculate the mass n of the solid in the component IV precipitate after removing ethanol3I.e. n3＝m3×x3；
When n is3＝M3When n is in the range, it is indicated that the impurity substance in the component IV is completely precipitated3＜M3In the meantime, it is indicated that the impurity substances in component IV are not completely precipitated and need to be continuously separated.
In one embodiment, the fraction V precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass of the solid in the fraction V precipitate after ethanol removal is n4The calculation method is as follows,
taking the mass as a after filter pressing4The precipitate of component V is evaporated to dryness at low temperature to obtain the product with mass b4The mass percentage content of the solid in the component V precipitate is calculated as x4Wherein, in the step (A),
then according to the mass percent content x4To calculate the mass n of the solid in the precipitate of component V after removal of ethanol4I.e. n4＝m4×x4；
When n is4＝M4When n is in the range, it indicates that the solid matter in the component V is completely precipitated4＜M4In the meantime, it is indicated that the solid matters in the component V are not completely precipitated and need to be continuously separated.
In one embodiment, the flow rates are controlled to be less than or equal to 1.0 liter/min during the preparation of component I, the preparation of components II + III, the preparation of component IV, the preparation of component V, and the purification of component V when the buffer is added.
In one embodiment, in the step of preparing the component I, 50% ethanol with the temperature lower than-15 ℃ is added until the volume ratio concentration of the ethanol is 8%; in the step of preparing the components II and III, 95 percent ethanol with the temperature lower than-15 ℃ is added until the volume concentration of the ethanol is 20 percent; in the step of preparing the component IV, 95 percent ethanol with the temperature lower than-15 ℃ is added until the volume concentration of the ethanol is 40 percent; in the step of refining component V, 25% ethanol at a temperature below-15 ℃ is added to a concentration of 10% by volume of ethanol.
In one embodiment, the flow rate of the ethanol added in the steps of preparing the component I, preparing the components II and III, preparing the component IV and refining the component V is 1.0-1.2 liters/minute.
In this example, in the steps of preparing the components II + III, preparing the component IV, preparing the component V and refining the component V, diatomaceous earth was added during pressure filtration, and the liquid inlet pressure was 0.20MPa or less.
In this example, in the step of preparing the components II + III, 18g of diatomaceous earth per liter of reaction solution was added for pressure filtration; in the step of preparing the component IV, 15g of diatomite is added into each liter of reaction liquid for filter pressing; in the step of preparing the component V, 4g of diatomite is added into each liter of reaction liquid for filter pressing; in the step of refining the component V, 20g of diatomaceous earth per liter of the reaction solution was added to carry out filter pressing.
The invention has the following beneficial effects:
the invention judges whether the corresponding steps meet the production requirements according to the precipitates obtained by pressure filtration in the steps of preparing the component I, preparing the component II + III, preparing the component IV and preparing the component V, because the components and the content thereof are stable in the production of the human serum albumin serving as the commercialized raw material plasma, namely the quality of the substances to be precipitated is also in a stable range in the steps of preparing the component I, preparing the component II + III, preparing the component IV and preparing the component V, the content of each main protein in the plasma is measured by sampling in the step of melting the plasma, and then the quality of the precipitated precipitates of each component is compared with the quality of the corresponding substances in the plasma in the steps of preparing the component I, preparing the component II + III, preparing the component IV and preparing the component V, so that whether the component is completely precipitated in the corresponding steps can be known, and whether the component meets the purification requirements is judged, the detection procedures in production are reduced, and the production efficiency is improved.
Drawings
FIG. 1 is a flow chart of the present invention.
Detailed Description
The invention is described in detail below with reference to the figures and examples.
It should be noted that all the directional terms such as "upper" and "lower" referred to herein are used with respect to the view of the drawings, and are only for convenience of description, and should not be construed as limiting the technical solution.
As shown in figure 1, an improved human serum albumin production process comprises plasma melting, detection, component I preparation, component II + III preparation, component IV preparation, component V refining, ultrafiltration, diluted preparation, pasteurization, protein sterilization, split charging and product incubation, wherein whether corresponding steps meet production requirements is judged according to precipitates obtained by pressure filtration in the steps of component I preparation, component II + III preparation, component IV preparation and component V preparation, and the specific production steps are as follows,
slurry melting: spraying raw plasma with 75% alcohol at a temperature lower than 30 ℃, washing the raw plasma with injection water at a temperature lower than 30 ℃ to remove the alcohol, drying the raw plasma, breaking the bag of the raw plasma, conveying the raw plasma to a plasma melting tank, performing interlayer circulation plasma melting with water at a temperature of 30-35 ℃, stopping water circulation in time after the plasma is melted, controlling the temperature of the plasma between 0-4 ℃, performing centrifugal separation and precipitation, and conveying the plasma after the precipitation is removed to a first reaction tank for extraction and separation;
and (3) detection: sampling from the reaction tank for detection, and respectively detecting the mass M of the substances to be separated in the steps of preparing the component I, preparing the components II and III, preparing the component IV and preparing the component V1、M2、M3And M4；
Preparing a component I: after plasma enters a first reaction tank, metering, starting stirring, controlling the temperature of the plasma to be between 0 and 1 ℃, dropwise adding a buffer solution with the pH value of 4.0, controlling the flow rate to be less than or equal to 1.0 liter/min when adding the buffer solution, adjusting the pH value to be between 6.80 and 7.00, adding 50 percent ethanol with the temperature of less than-15 ℃ until the volume ratio concentration of the ethanol is 8 percent, adding the ethanol at the flow rate of 1.0 to 1.2 liters/min, controlling the temperature to be between-1 and-3 ℃ after adding the ethanol, centrifuging after adding the ethanol, obtaining a component I precipitate and a component I supernatant through centrifuging, weighing the mass of the component I precipitate to be m1Comparison of the component I precipitation masses m1Mass M of substance to be separated from component I1If the mass m of the component I precipitate1Mass M of substance to be separated greater than component I1Then the supernatant of the component I enters the next working procedure, if the mass m of the precipitation of the component I1Is less than or equal toMass M of the substance to be separated of component I1Adjusting the production parameters of the first reaction tank, adjusting the input amount of low-temperature ethanol to increase the mass of the component I precipitate, performing filter pressing again, and comparing the total mass m of the component I precipitate1Mass M of substance to be separated from component I1Repeating the operation until the component I precipitate is completely separated out, and allowing the component I supernatant to enter the next step, wherein the component I precipitate is used for preparing human fibrinogen;
preparing a component II + III: taking the supernatant of the component I into a second reaction tank, dropwise adding a buffer solution with the pH of 4.0 to adjust the pH value of the supernatant of the component I to be 6.80-6.85, controlling the flow rate to be less than or equal to 1.0 liter/min when adding the buffer solution, adding 95% ethanol with the temperature of less than-15 ℃ until the volume ratio concentration of the ethanol is 20%, adding the ethanol at the flow rate of 1.0-1.2 liters/min, controlling the temperature to be-4 to-6 ℃ after adding the ethanol, controlling the pH to be 6.80-7.00 after adding the ethanol, stirring for more than 120 minutes, standing for more than 60 minutes, starting stirring, adding 18g of diatomite into each liter of reaction liquid for filter pressing, obtaining the component II + III sediment and the supernatant of the component II + III sediment after filter pressing, and weighing the mass of the component II + III sediment to be m2Comparison of the component II + III precipitation masses m2Mass M of substance to be separated from Components II + III2If the mass m of the component II + III precipitates2Mass M of substance to be separated greater than components II + III2Then the supernatant of the component II + III enters the next working procedure, if the mass m of the component II + III precipitate is2Mass M less than or equal to mass of substance to be separated of Components II + III2Adjusting the production parameters of the second reaction tank, adjusting the input amount of low-temperature ethanol to increase the mass of the component II + III precipitate, performing filter pressing again, and comparing the total mass m of the component II + III precipitate2Mass M of substance to be separated from Components II + III2Repeating the operation until the component II + III precipitates are completely separated out, and allowing the component II + III supernatant to enter the next step, wherein the component II + III precipitates are used for preparing immunoglobulin;
preparing a component IV: taking component II + III supernatant fluid into a third reaction tank, dropwise adding a buffer solution with the pH of 4.0 to adjust the pH of the component II + III supernatant fluidThe value is 5.70-5.90, the flow rate is controlled to be less than or equal to 1.0 liter/min when buffer solution is added, the temperature in a third reaction tank is controlled to be-4-6 ℃, 95% ethanol with the temperature lower than-15 ℃ is added until the volume concentration of the ethanol is 40%, the flow rate of the added ethanol is 1.0-1.2 liters/min, the pH value of the liquid is 5.90-6.10 after the ethanol is added, the liquid is stirred for more than 120 minutes, the stirring is started for more than 60 minutes, 15g of diatomite is added into each liter of reaction liquid for filter pressing, the liquid inlet pressure is not more than 0.20MPa at most, component IV sediment and component IV supernatant are obtained after the filter pressing, and the mass of the component IV sediment is weighed to be m3Comparison of the precipitated masses m of component IV3Mass M of substance to be separated from component IV3If component IV precipitates by mass m3Mass M of substance to be separated greater than component IV3The supernatant of the component IV enters the next working procedure, and if the component IV precipitates by mass m3Mass M less than or equal to that of the substance to be separated of component IV3Adjusting the production parameters of the third reaction tank, adjusting the input amount of low-temperature ethanol to increase the mass of the component I precipitate, performing filter pressing again, and comparing the total mass m of the component IV precipitate3Mass M of substance to be separated from component IV3Repeating the operation until the component IV precipitate is completely separated out, and allowing the component IV supernatant to enter the next step, wherein the component IV precipitate is discarded;
preparing a component V: taking the supernatant of the component IV into a fourth reaction tank, dropwise adding a buffer solution with the pH of 4.0 to adjust the pH value of the supernatant of the component IV to be 4.80-5.00, controlling the flow rate to be less than or equal to 1.0 liter/min when the buffer solution is added, controlling the temperature in the fourth reaction tank to be below-8 ℃, stirring for more than 120 min, then adding 4g of diatomite into each liter of reaction liquid, performing filter pressing, wherein the pressure of the liquid inlet is not more than 0.20MPa at most, obtaining a component V precipitate and a component V supernatant after the filter pressing, and weighing the mass of the component V precipitate to be m4Comparison of the Mass m of the component V precipitate4Mass M of substance to be separated from component V4If component V precipitates a mass m4Mass M of the substance to be separated greater than component V4Then the component V is precipitated and enters the next working procedure, if the component V is precipitated by mass m4Mass M of the substance to be separated of fraction V or less4Adjusting the production parameters of the fourth reaction tank to increase the mass of the component V precipitate, and comparing the total mass m of the component V precipitate after filter pressing again4Mass M of substance to be separated from component V4Repeating the operation until the component V precipitate is completely separated out, and entering the next step, wherein the component V supernatant is transferred to an alcohol recovery tower to recover alcohol;
refining a component V: taking a component V precipitate, fully dissolving the component V precipitate with 5 times of injection water, dropwise adding a buffer solution with the pH value of 4.0 to adjust the pH value of a component V precipitate dissolving solution to be 4.50-4.60, controlling the flow rate to be less than or equal to 1.0 liter/min when adding the buffer solution, adding 25% ethanol with the temperature of less than-15 ℃ until the volume ratio concentration of the ethanol is 10%, adding the ethanol with the flow rate of 1.0-1.2 liters/min (injection: 1kg of the component V precipitate is equivalent to 1L of the 30% ethanol-containing precipitate dissolving solution), controlling the temperature to be-2-3 ℃, stirring for more than 120 minutes, adding 20g of diatomite into each liter of reaction solution, carrying out filter pressing, wherein the highest liquid inlet pressure is not more than 0.20MPa, filtering the filtrate by using a Zetaplus deep filter core to obtain a component V purified solution, and the highest liquid inlet pressure is not more than 0.20MPa in the filtering process;
and (3) ultrafiltration: controlling the inlet hydraulic pressure to be not more than 0.45Mpa, the reflux pressure to be not more than 0.30Mpa and the pressure difference between the inlet hydraulic pressure and the reflux pressure to be more than 0.10Mpa in the ultrafiltration process, concentrating the component V purified solution until the protein content is more than 15%, carrying out isovolumetric dialysis by sequentially using 4 times of volume of 0.85% sodium chloride solution at the temperature of 2-8 ℃ and 5 times of volume of 0.5% sodium chloride solution at the temperature of 2-8 ℃ after the concentration is finished, concentrating the protein concentration to be more than 20% after the dialysis is finished, and adjusting the pH value of the protein solution to be 6.90-7.10 by using 1mol/L NaHCO 3;
diluting and preparing: diluting according to 20% protein concentration, respectively adding sodium caprylate as protective agent to 0.16mmol/g protein concentration and sodium chloride to 145mmol/L final stock solution, adding water for injection to the final diluted amount, and stirring well.
And (3) pasteurizing inactivation: after the diluted preparation solution is uniformly stirred, jacket circulating water at 60.5 ℃ is used for heating, when the temperature of the protein solution rises to 59.5 ℃, timing is started, inactivation is carried out at 59.5-60.5 ℃ for 12 hours, and after the inactivation is finished, the temperature of the protein solution is reduced to 20-30 ℃.
Protein sterilization, subpackaging and product incubation: and (3) sterilizing and filtering the inactivated protein liquid by using a 0.2-micron sterilizing filter element, filling according to requirements, transferring the filled product to a hatching room at the temperature of 30-32 ℃, incubating for 14 days, and performing finished product verification after incubation.
Packaging a finished product: and packaging the qualified products.
Wherein, the reagents required in the production process are as follows:
ph4.0 buffer: 65.6g of anhydrous sodium acetate is fully dissolved by using a proper amount of water for injection, 244.9ml of glacial acetic acid is added, and the water for injection is added to 1L and mixed evenly.
1mol/L sodium bicarbonate solution: dissolving 84g of sodium bicarbonate with appropriate amount of water for injection, adding water for injection to 1L, and mixing.
50% ethanol solution: 0.474kg of 95% ethanol is taken, and water for injection is added until the volume is 1kg, and the mixture is stirred evenly.
25% ethanol solution: 0.220kg of 95% ethanol is taken, and water for injection is added until the volume is 1kg, and the mixture is uniformly stirred.
0.85% sodium chloride dialysate: 8.5g of sodium chloride is fully dissolved by using a proper amount of water for injection, and the water for injection is added to 1L and stirred uniformly.
0.5% sodium chloride dialysate: 5g of sodium chloride is fully dissolved by using a proper amount of water for injection, and the water for injection is added to 1L and stirred uniformly.
In this example, the fraction I precipitate still contained a small amount of ethanol solution after pressure filtration, wherein the mass of the solid in the fraction I precipitate after removal of ethanol was n1The calculation method is as follows,
taking the mass as a after filter pressing1The precipitate of component I is evaporated to dryness at low temperature to obtain the product with mass b1The mass percentage content of the solid in the component I precipitate is calculated as x1Wherein, in the step (A),
then according to the mass percent content x1To calculate the mass n of the solid in the component I precipitate after removing ethanol1I.e. n1＝m1×x1；
When n is1＝M1When n is less than n, the impurity substance in the component I is completely precipitated1＜M1In this case, it was shown that the impurity substances in component I were not completely precipitated.
In this example, the fraction II + III precipitate still contained a small amount of ethanol solution after pressure filtration, wherein the mass n of the solid from which ethanol was removed in the fraction II + III precipitate was n2The calculation method is as follows,
taking the mass as a after filter pressing2The precipitate of the components II and III is evaporated to dryness at low temperature to obtain the product with the mass b2The mass percentage of the solid in the component II + III precipitate is calculated as x2Wherein, in the step (A),
then according to the mass percent content x2To calculate the mass n of the solid in the component II + III precipitate after removing ethanol2I.e. n2＝m2×x2；
When n is2＝M2When n is less than n, the impurity substances in the components II + III are completely separated out2＜M2When the impurity substances in the components II + III are not completely separated out.
In this example, the fraction IV precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass of the solid in the fraction IV precipitate after removal of ethanol is n3The calculation method is as follows,
taking the mass as a after filter pressing3The precipitate of component IV is evaporated to dryness at low temperature to obtain the product with mass b3The mass percentage content of the solid in the component IV precipitate is calculated as x3Wherein, in the step (A),
then according to the mass percent content x3To calculate the mass n of the solid in the component IV precipitate after removing ethanol3I.e. n3＝m3×x3；
When n is3＝M3When n is in the range, it is indicated that the impurity substance in the component IV is completely precipitated3＜M3In this case, it was found that the impurity substances in the component IV were not completely precipitated.
In this example, the fraction IV precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass of the solid in the fraction IV precipitate after removal of ethanol is n3The calculation method is as follows,
taking the mass as a after filter pressing3The precipitate of component IV is evaporated to dryness at low temperature to obtain the product with mass b3The mass percentage content of the solid in the component IV precipitate is calculated as x3Wherein, in the step (A),
then according to the mass percent content x3To calculate the mass n of the solid in the component IV precipitate after removing ethanol3I.e. n3＝m3×x3；
When n is3＝M3When n is in the range, it is indicated that the impurity substance in the component IV is completely precipitated3＜M3In this case, it was found that the impurity substances in the component IV were not completely precipitated.
In this example, fraction V precipitate still contained a small amount of ethanol solution after pressure filtration, wherein the mass of the solid in fraction V precipitate after removal of ethanol was n4The calculation method is as follows,
taking the mass as a after filter pressing4The precipitate of component V is evaporated to dryness at low temperature to obtain the product with mass b4The mass percentage content of the solid in the component V precipitate is calculated as x4Wherein, in the step (A),
then according to the mass percent content x4To calculate the mass n of the solid in the precipitate of component V after removal of ethanol4I.e. n4＝m4×x4；
When n is4＝M4When n is in the range, it indicates that the solid matter in the component V is completely precipitated4＜M4In this case, it is indicated that the solid matter in component V is not completely precipitated.
The above description is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may occur to those skilled in the art without departing from the principle of the invention, and are considered to be within the scope of the invention.
Claims (10)
1. An improved human serum albumin production process is characterized by comprising plasma melting, detection, component I preparation, component II + III preparation, component IV preparation, component V refining, ultrafiltration, diluted preparation, pasteurization, split charging and product incubation, wherein whether corresponding steps meet production requirements or not is judged according to precipitates obtained by filter pressing in the steps of component I preparation, component II + III preparation, component IV preparation and component V preparation, and the specific operation is as follows,
and (3) detection: sampling from the reaction tank for detection, and respectively detecting the mass M of the substances to be separated in the steps of preparing the component I, preparing the components II and III, preparing the component IV and preparing the component V1、M2、M3And M4；
Preparing a component I: after the plasma enters a first reaction tank, dropwise adding a buffer solution to adjust the pH value, adding low-temperature ethanol, centrifuging after adding the ethanol, centrifuging to obtain a component I precipitate and a component I supernatant, and weighing the component I precipitate to obtain the mass m1Comparison of the component I precipitation masses m1Mass M of substance to be separated from component I1If the mass m of the component I precipitate1Mass M less than or equal to that of the substance to be separated of component I1Adjusting the production parameters of the first reaction tank to increase the mass of the component I precipitate, and comparing the total mass m of the component I precipitate after filter pressing again1Mass M of substance to be separated from component I1Repeating the operation until the component I is completely precipitated, and allowing the component I supernatant to enter the next step;
preparing a component II + III: taking the supernatant of the component I into a second reaction tank, dropwise adding a buffer solution to adjust the pH value of the supernatant of the component I, and adding low-temperature ethanolAfter the ethanol is added, stirring and standing, starting stirring, then performing filter pressing, obtaining component II + III precipitates and component II + III supernatant after filter pressing, and weighing the mass m of the component II + III precipitates2Comparison of the component II + III precipitation masses m2Mass M of substance to be separated from Components II + III2If the mass m of the component II + III precipitates2Mass M less than or equal to mass of substance to be separated of Components II + III2Adjusting the production parameters of the second reaction tank to increase the mass of the component II + III precipitate, and comparing the total mass m of the component II + III precipitate after pressure filtration again2Mass M of substance to be separated from Components II + III2Repeating the operation until the component II + III precipitates are completely separated out, and enabling the component II + III supernatant to enter the next working procedure;
preparing a component IV: taking component II + III supernatant fluid into a third reaction tank, dropwise adding a buffer solution to adjust the pH value of the component II + III supernatant fluid, adding low-temperature ethanol, stirring, standing, starting stirring, then performing filter pressing, obtaining component IV precipitate and component IV supernatant fluid after filter pressing, and weighing the mass of the component IV precipitate as m3Comparison of the precipitated masses m of component IV3Mass M of substance to be separated from component IV3If component IV precipitates by mass m3Mass M less than or equal to that of the substance to be separated of component IV3Adjusting the production parameters of the third reaction tank to increase the mass of the component I precipitate, and comparing the total mass m of the component IV precipitate after pressure filtration again3Mass M of substance to be separated from component IV3Repeating the operation until the component IV precipitate is completely separated out, and allowing the component IV supernatant to enter the next step;
preparing a component V: taking the supernatant of the component IV into a fourth reaction tank, dropwise adding a buffer solution to adjust the pH value of the supernatant of the component IV, stirring, performing filter pressing, and obtaining a component V precipitate and a component V supernatant after filter pressing, wherein the mass of the component V precipitate is weighed to be m4Comparison of the Mass m of the component V precipitate4Mass M of substance to be separated from component V4If component V precipitates a mass m4Mass M of the substance to be separated of fraction V or less4Adjusting the production parameters of the fourth reaction tankIncreasing the mass of the component V precipitate, and comparing the total mass m of the component V precipitate after filter pressing again4Mass M of substance to be separated from component V4And (4) repeating the operation until the component V is completely precipitated, and enabling the component V to be precipitated to enter the next step, wherein the component V supernatant is transferred to an alcohol recovery tower to recover alcohol.
2. The improved process for the production of human serum albumin of claim 1, wherein fraction I precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass of the fraction I precipitate after ethanol removal from solids is n1The calculation method is as follows,
taking the mass as a after filter pressing1The precipitate of component I is evaporated to dryness at low temperature to obtain the product with mass b1The mass percentage content of the solid in the component I precipitate is calculated as x1Wherein, in the step (A),
then according to the mass percent content x1To calculate the mass n of the solid in the component I precipitate after removing ethanol1I.e. n1＝m1×x1；
When n is1＝M1When n is less than n, the impurity substance in the component I is completely precipitated1＜M1In the meantime, it is indicated that the impurity substances in the component I are not completely precipitated and need to be continuously separated.
3. The improved process for the production of human serum albumin of claim 1, wherein the fraction II + III precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass n of the fraction II + III precipitate after ethanol removal from the solids is n2The calculation method is as follows,
taking the mass as a after filter pressing2The precipitate of the components II and III is evaporated to dryness at low temperature to obtain the product with the mass b2The mass percentage of the solid in the component II + III precipitate is calculated as x2Wherein, in the step (A),
then according to the mass percent content x2To calculate the mass n of the solid in the component II + III precipitate after removing ethanol2I.e. n2＝m2×x2；
When n is2＝M2When n is less than n, the impurity substances in the components II + III are completely separated out2＜M2In the meantime, it is indicated that the impurity substances in the components II + III are not completely separated out and need to be continuously separated.
4. The improved process for the production of human serum albumin of claim 1, wherein fraction IV precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass of the fraction IV precipitate after ethanol removal from solids is n3The calculation method is as follows,
taking the mass as a after filter pressing3The precipitate of component IV is evaporated to dryness at low temperature to obtain the product with mass b3The mass percentage content of the solid in the component IV precipitate is calculated as x3Wherein, in the step (A),
then according to the mass percent content x3To calculate the mass n of the solid in the component IV precipitate after removing ethanol3I.e. n3＝m3×x3；
When n is3＝M3When n is in the range, it is indicated that the impurity substance in the component IV is completely precipitated3＜M3In the meantime, it is indicated that the impurity substances in component IV are not completely precipitated and need to be continuously separated.
5. The improved process for the production of human serum albumin of claim 1, wherein fraction V precipitate still contains a small amount of ethanol solution after pressure filtration, wherein the mass of the fraction V precipitate after ethanol removal from solids is n4The calculation method is as follows,
taking the mass as a after filter pressing4The precipitate of component V is evaporated to dryness at low temperature to obtain the product with mass b4The mass percentage content of the solid in the component V precipitate is calculated as x4Wherein, in the step (A),
then according to the mass percent content x4To calculate the mass n of the solid in the precipitate of component V after removal of ethanol4I.e. n4＝m4×x4；
When n is4＝M4When n is in the range, it indicates that the solid matter in the component V is completely precipitated4＜M4In the meantime, it is indicated that the solid matters in the component V are not completely precipitated and need to be continuously separated.
6. The improved process for the production of human serum albumin of any of claims 1-5, wherein the buffer is added at a controlled flow rate of less than or equal to 1.0 liter/min for each of component I, component II + III, component IV, component V and component V purification.
7. The improved process for the production of human serum albumin of any of claims 1-5, wherein in the step of preparation of component I, 50% ethanol at a temperature below-15 ℃ is added to a concentration of 8% by volume of ethanol; in the step of preparing the components II and III, 95 percent ethanol with the temperature lower than-15 ℃ is added until the volume concentration of the ethanol is 20 percent; in the step of preparing the component IV, 95 percent ethanol with the temperature lower than-15 ℃ is added until the volume concentration of the ethanol is 40 percent; in the step of refining component V, 25% ethanol at a temperature below-15 ℃ is added to a concentration of 10% by volume of ethanol.
8. The improved process for producing human serum albumin of claim 1, wherein the ethanol is added at a flow rate of 1.0 to 1.2 liters/minute in the steps of component I preparation, component II + III preparation, component IV preparation and component V purification.
9. The improved process for producing human serum albumin as claimed in claim 1, wherein in the steps of preparing component II + III, preparing component IV, preparing component V and refining component V, diatomaceous earth is added during pressure filtration, and the inlet pressure is less than or equal to 0.20 MPa.
10. The improved process for producing human serum albumin as claimed in claim 9, wherein in the step of preparing the components II + III, 18g of diatomaceous earth is added per liter of the reaction solution to carry out filter pressing; in the step of preparing the component IV, 15g of diatomite is added into each liter of reaction liquid for filter pressing; in the step of preparing the component V, 4g of diatomite is added into each liter of reaction liquid for filter pressing; in the step of refining the component V, 20g of diatomaceous earth per liter of the reaction solution was added to carry out filter pressing.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113773381A (en) * | 2021-09-27 | 2021-12-10 | 广东丹霞生物制药有限公司 | Improved production method of human immunoglobulin |
| CN113801222A (en) * | 2021-09-27 | 2021-12-17 | 广东丹霞生物制药有限公司 | Method for producing real-time monitored human immune globulin |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102311496A (en) * | 2011-08-22 | 2012-01-11 | 西安回天血液制品有限责任公司 | Method for recovering alhumin from deposited components I+II+III from cold ethanol method |
| CN103394076A (en) * | 2013-08-15 | 2013-11-20 | 江西博雅生物制药股份有限公司 | Process for preparing human serum albumin |
| CN103709245A (en) * | 2013-12-20 | 2014-04-09 | 华兰生物工程重庆有限公司 | Method for low-temperature extraction of human serum albumin employing ethanol |
| CN104004084A (en) * | 2014-04-12 | 2014-08-27 | 浙江海康生物制品有限责任公司 | Producing method of human serum albumin |
| CN104558156A (en) * | 2015-01-23 | 2015-04-29 | 郑州莱士血液制品有限公司 | Method for extracting human serum albumin from plasma and increasing yield |
| CN107857811A (en) * | 2017-12-20 | 2018-03-30 | 广东丹霞生物制药有限公司 | A kind of preparation technology of human serum albumin |
| CN110305208A (en) * | 2018-03-27 | 2019-10-08 | 发贵科技（贵州）有限公司 | The human serum albumin of cold ethanol two-step method separation |
| CN110317262A (en) * | 2018-03-28 | 2019-10-11 | 发贵科技（贵州）有限公司 | Cold ethanol two-step method human serum albumin separating technology |
-
2020
- 2020-12-22 CN CN202011535109.7A patent/CN112521487A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102311496A (en) * | 2011-08-22 | 2012-01-11 | 西安回天血液制品有限责任公司 | Method for recovering alhumin from deposited components I+II+III from cold ethanol method |
| CN103394076A (en) * | 2013-08-15 | 2013-11-20 | 江西博雅生物制药股份有限公司 | Process for preparing human serum albumin |
| CN103709245A (en) * | 2013-12-20 | 2014-04-09 | 华兰生物工程重庆有限公司 | Method for low-temperature extraction of human serum albumin employing ethanol |
| CN104004084A (en) * | 2014-04-12 | 2014-08-27 | 浙江海康生物制品有限责任公司 | Producing method of human serum albumin |
| CN104558156A (en) * | 2015-01-23 | 2015-04-29 | 郑州莱士血液制品有限公司 | Method for extracting human serum albumin from plasma and increasing yield |
| CN107857811A (en) * | 2017-12-20 | 2018-03-30 | 广东丹霞生物制药有限公司 | A kind of preparation technology of human serum albumin |
| CN110305208A (en) * | 2018-03-27 | 2019-10-08 | 发贵科技（贵州）有限公司 | The human serum albumin of cold ethanol two-step method separation |
| CN110317262A (en) * | 2018-03-28 | 2019-10-11 | 发贵科技（贵州）有限公司 | Cold ethanol two-step method human serum albumin separating technology |
Non-Patent Citations (1)
| Title |
|---|
| 张安山: "实现自控分离对人血浆蛋白质量的影响", 《中国生物制品学杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113773381A (en) * | 2021-09-27 | 2021-12-10 | 广东丹霞生物制药有限公司 | Improved production method of human immunoglobulin |
| CN113801222A (en) * | 2021-09-27 | 2021-12-17 | 广东丹霞生物制药有限公司 | Method for producing real-time monitored human immune globulin |
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Application publication date: 20210319 |