CN110054685A - A kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III - Google Patents
A kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III Download PDFInfo
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- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
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Abstract
The present invention relates to human immunoglobulin(HIg) field more particularly to a kind of production methods of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III.It comprises the following steps: II+III precipitate production method of F；Control dissolution F II+III precipitates dissolution water and lysate temperature；The control of sodium acetate solution temperature is added at 0 ~ 4.0 DEG C；Adjust pH；It is added before ethyl alcohol and lysate cooling is precipitated to F II+III；Filter aid diatomite is added to be separated after stirring 15 minutes with filter press, obtains FII precipitating and FIII supernatant；It is 0.9794 ~ 0.9746 g/cm that densimeter method, which detects III supernatant density of F,3, it is 3.8 ~ 5.6g/L that biuret method, which detects protein content, and FIII Sediment weight scope control is throwing slurry amount × (0.07419 ± 3 × 0.00267) kg.
Description
Technical field
The present invention relates to human immunoglobulin(HIg) field more particularly to a kind of intravenous human immunoglobulin(HIg) (pH4) cold ethanols
The production method of component III.
Background technique
Cold ethanol method is one of preparation method of human plasma protein fraction, and this method is to be cured by Harvard University for 1940
The Edwin Cohn teaching inventive of institute.Cold ethanol method is to reduce acidity step by step using pooled plasma as raw material.Improve second
Determining alcohol, while temperature is reduced, substep is precipitated various albumen from solution in the form of component under different conditions, and passes through
It is separated by filtration out.Ethyl alcohol has many advantages as a kind of protein precipitant: dielectric constant is low, with water miscible, normal
Under environment and operating condition without explosion hazard, low molecular weight, relative inertness chemically, low toxicity, it is inexpensive, be easy to get and bacteriostasis.
So far, extensive plasma protein separation still uses cold ethanol partition method substantially.
Summary of the invention
The purpose of invention: in order to provide a kind of better intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III of effect
Production method, specific purposes see specific implementation part multiple substantial technological effects.
In order to reach purpose as above, the present invention is adopted the following technical scheme that:
A kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III, which is characterized in that include following step
It is rapid:
(1) II+III precipitate production method of F: refrigerated plasma with dedicated buffering liquid adjusts blood plasma pH value to 7.15 ~ 7.25 after melting,
Dedicated buffering liquid is acetic acid/sodium acetate solution；Add -15 DEG C of 95% ethyl alcohol below into reactor tank, until ethyl alcohol final concentration in tank
It is 8%, blood plasma temperature is controlled at -1.5 ~ -2.5 DEG C；Filters pressing separation obtains component I, that is, FI precipitating and FI supernatant, into FI supernatant
The mixed liquor of dedicated buffering liquid and ethyl alcohol is added, makes ethyl alcohol final concentration of 22%, -5.0 ~ -6.0 DEG C of liquid temperature, it is small to continue stirring 2.5
When, surveying pH value is 6.70 ~ 6.80, otherwise with dedicated buffering liquid or the NaHCO of 0.872mol/L3Filter aid pearl is added in adjustment
Rock, diatomite are separated after stirring 45 minutes with filter press, collect component, which is divided into the precipitating of F II+III；
(2) control dissolution F II+III precipitates dissolution water and lysate temperature is 0 ~ 4.0 DEG C, and mixing time is 2 ~ 4 hours；
(3) control of sodium acetate solution temperature is added at 0 ~ 4.0 DEG C；The timing since be initially added into sodium acetate solution, time control
In 4 ~ 5 hours, mixing time control was at 1 ~ 2 hour after addition；
(4) when adjusting pH, uniformly being mixed with both dedicated buffering liquid and water for injection for pH will be adjusted and be added, mixed liquor is controlled
Temperature is to 0 ~ 4.0 DEG C；The liquid feeding time controlled in 2 ~ 3 hours, and suspension pH value is controlled 5.15 ~ 5.25；
(5) it is added before ethyl alcohol and lysate cooling is precipitated to F II+III, temperature is down to 0 ~ 1.0 DEG C in one hour；
(6) be added concentration of alcohol be 95%, temperature control at -15.0 DEG C hereinafter, from be added ethyl alcohol timing, from addition second
Alcohol starts timing, and the liquid feeding time controlled at 3 ~ 5 hours, and lysate temperature is down to -5.0 ~ -6.0 by ethyl alcohol final concentration of 17%
℃；Suspension pH value should be 5.10 ~ 5.30 after ethyl alcohol is added；
(7) filter aid diatomite is added to be separated after stirring 15 minutes with filter press, obtains component FII) it precipitates and FIII supernatant；
(8) densimeter method detection III supernatant density of F is 0.9794 ~ 0.9746 g/cm3, biuret method detection protein content be
3.8 ~ 5.6g/L, FIII Sediment weight scope control are throwing slurry amount × (0.07419 ± 3 × 0.00267) kg.
The further technical solution of the present invention is, the producer of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III
Method are as follows:
The dissolution that F II+III is precipitated in step (2): II+III theoretical weight (A) of F for not adding diatomite, formula: A=blood plasma are calculated
It measures (kg) × 0.04；The amount that dissolution F II+III precipitates water for injection used is calculated, formula: injection water (kg) needed for dissolving=
Plasma volume (kg) × 0.6405-F II+III precipitates (kg)；Dissolving F II+III to precipitate the temperature of water for injection used is 0 ~ 4 DEG C, control
It is 0 ~ 4.0 DEG C that F II+III processed, which precipitates lysate temperature,；It precipitates within stirring 2 ~ 4 hours and is completely dissolved to F II+III.
The further technical solution of the present invention is, sodium acetate solution is added in step (3): calculating and sodium acetate solution is added
Amount, formula: NaAc 3H2O(kg) = A×0.054；Injection water (kg)=constant A × 2；Sodium acetate solution amount
(kg)=plasma volume (kg) × 0.08216, sodium acetate solution temperature are controlled at 0 ~ 4.0 DEG C；By sodium acetate solution at 4 ~ 5 hours
Interior constant speed is added in II+III lysate of F, continues stirring 1 ~ 2 hour.
The further technical solution of the present invention is, buffer is added in step (4) and adjusts pH: calculating and dedicated buffering is added
The amount of liquid, formula: dedicated buffering liquid additional amount (kg)=(F II+III precipitation capacity (kg)+0.24 × plasma volume (kg)+sodium acetate
Amount of solution (kg)) × 0.0043, dedicated buffering liquid dilution injection water (kg)=constant A, by dedicated buffering liquid and injection
It is uniformly mixed with both water, controls mixeding liquid temperature to 0 ~ 4.0 DEG C；It is small 2 ~ 3 by dedicated buffering liquid and water for injection mixed liquor
When interior constant speed be added in reactor tank, adjust in reactor tank suspension pH value to 5.15 ~ 5.25.
The further technical solution of the present invention is, ethyl alcohol is added in step (6): calculating and amount of alcohol: formula is added: being added
Suspension total amount (kg) × 0.180 in amount of alcohol (kg)=reactor tank；Cool down to product, temperature is down to 0 ~ 1.0 DEG C in one hour；
By 95% ethyl alcohol (- 15.0 DEG C or less), constant speed is added in reactor tank in 3 ~ 5 hours, until the ethyl alcohol of suspension final concentration of 17%,
Temperature is down to -5.0 ~ -6.0 DEG C；It finishes and continues to sample for stirring 10 minutes, survey suspension pH value, should be 5.10 ~ 5.30, such as exceed
Range is adjusted with dedicated buffering liquid or 0.872mol/L NaHCO3 liquid.
The further technical solution of the present invention is that filters pressing separates in step (7) a metallic, obtains III supernatant F III of F precipitating, F III
Supernatant continues separation component II, uses densitometer detection III supernatant density of F for 0.9794 ~ 0.9746 g/cm3, using biuret
Method detect protein content be 3.8 ~ 5.6g/L, FIII Sediment weight scope control throw slurry amount × (0.07419 ± 3 ×
0.00267) kg.
The further technical solution of the present invention is that the product variety that this technique is suitable for production includes: intravenous people is immune
Globulin (pH4) liquid and freeze-dried formulation each specification [0.5g/ bottles (5 %, 10ml), lg/ bottles (5 %, 20ml), 1.
25g/ bottles (5 %, 25ml), 2. 5 g/bottle (5 %, 50ml), 5g/ bottles (5%, 100ml), 10g/ bottles (5%,
200ml)] and human immunoglobulin(HIg) class product.
Using the present invention of technical solution as above, have the advantages that compared with the existing technology: by carefully studying and
Many experiments, discovery influence cold ethanol method protein precipitation reaction factor it is main there are five, it may be assumed that pH value, temperature, protein compression
Degree, ionic strength and concentration of alcohol.Intravenous human immunoglobulin(HIg) (pH4) product is in cold ethanol method production process due to influencing
The selection of five technological parameters of protein precipitation reaction influences intravenous human immunoglobulin(HIg) (pH4) yield and product quality refers to
Mark.By control F II+III precipitate dissolution conditions, dissolution injection water, be added sodium acetate promote IgG dissolution, adjustment pH,
The method and monitoring III supernatant density of F, III Sediment weight of albumen and F of ethyl alcohol is added, accurately controls intravenous people's immune globulin
The production process of white (pH4) cold ethanol component III, it can be ensured that intravenous human immunoglobulin(HIg) (pH4) high income, quality index
It is good.
Detailed description of the invention
Fig. 1 is the situation of change that F II+III precipitates protein content in course of dissolution；
Fig. 2 is that table 1 is intravenous human immunoglobulin(HIg) (pH4) quality index situation.
Specific embodiment
With reference to embodiment, the present invention is furture elucidated, it should be understood that following specific embodiments are only used for
It is bright the present invention rather than limit the scope of the invention.This patent provides a variety of concomitant regimens, and different expression place belongs to and is based on
The modified scheme of basic scheme either parallel type scheme.Every kind of scheme has the unique features of oneself.
Present invention employs the characteristic technology controlling and process means of six aspects
(1) control dissolution F II+III precipitates dissolution water and lysate liquid temperature is 0 ~ 4.0 DEG C, and mixing time is 2 ~ 4 hours.
(2) control of sodium acetate solution temperature is added at 0 ~ 4.0 DEG C.The liquid feeding time controlled in 4 ~ 5 hours, stirred after addition
Time control is mixed at 1 ~ 2 hour.
(3) when adjusting pH, by adjustment pH, with both dedicated buffering liquid and water for injection, uniformly mixing is added, control mixing
Liquid temperature is to 0 ~ 4.0 DEG C.The liquid feeding time controlled in 2 ~ 3 hours, and suspension pH value is controlled 5.15 ~ 5.25.
(4) cool down before ethyl alcohol is added to product, temperature is down to 0 ~ 1.0 DEG C in 1 hour.
(5) concentration of alcohol being added is 95%, temperature control at -15.0 DEG C hereinafter, the control of liquid feeding time was at 3 ~ 5 hours, second
Alcohol final concentration of 17%, temperature is down to -5.0 ~ -6.0 DEG C.Suspension pH value should be 5.10 ~ 5.30 after ethyl alcohol is added.
(6) densimeter method detection III supernatant density of F is 0.9794 ~ 0.9746 g/cm3, biuret method detection protein contain
Amount is 3.8 ~ 5.6g/L, and FIII Sediment weight scope control is throwing slurry amount × (0.07419 ± 3 × 0.00267) kg.
On the basis of above-mentioned six characteristic steps, a kind of intravenous human immunoglobulin(HIg) (pH4) low temperature second of the invention
The production method of alkoxide component III includes:
(1) dissolution that F II+III is precipitated: II+III theoretical weight (A) of F for not adding diatomite is calculated, formula: A=plasma volume (kg) ×
0.04。
Calculate the amount that dissolution F II+III precipitates water for injection used, formula: injection water (kg)=blood plasma needed for dissolving
It measures (kg) × 0.6405-F II+III and precipitates (kg).Dissolving F II+III to precipitate the temperature of water for injection used is 0 ~ 4 DEG C, controls F
II+III precipitating lysate temperature is 0 ~ 4.0 DEG C.It precipitates within stirring 2 ~ 4 hours and is completely dissolved to F II+III.
(2) sodium acetate solution is added: calculating the amount that sodium acetate solution is added, formula: NaAc 3H2O (kg)=constant A
×0.054；Injection water (kg)=constant A × 2；Sodium acetate solution amount (kg)=plasma volume (kg) × 0.08216, acetic acid
Sodium solution temperature is controlled at 0 ~ 4.0 DEG C.By sodium acetate solution, constant speed is added in II+III lysate of F in 4 ~ 5 hours, continues to stir
It mixes 1 ~ 2 hour.
(3) buffer is added and adjusts pH: calculating the amount that dedicated buffering liquid is added, formula: dedicated buffering liquid additional amount (kg)
=(II+III+0.24 × plasma volume of precipitation capacity kg (kg) of F+sodium acetate solution amount (kg)) × 0.0043, the dilution of dedicated buffering liquid
With injection water (kg)=A, both dedicated buffering liquid and water for injection are uniformly mixed, control mixeding liquid temperature to 0 ~ 4.0
℃.By dedicated buffering liquid and water for injection mixed liquor, constant speed is added in reactor tank in 2 ~ 3 hours, adjusts suspension in reactor tank
PH value is to 5.15 ~ 5.25.
(4) be added ethyl alcohol: calculate be added amount of alcohol: formula: be added amount of alcohol (kg)=reactor tank in suspension total amount (kg) ×
0.180.Cool down to product, temperature is down to 0 ~ 1.0 DEG C in 1 hour.By 95% ethyl alcohol (- 15.0 DEG C or less) in 3 ~ 5 hours
Constant speed is added in reactor tank, until the ethyl alcohol of suspension final concentration of 17%, temperature is down to -5.0 ~ -6.0 DEG C.It finishes and continues stirring 10
Minute sampling, surveys suspension pH value, should be 5.10 ~ 5.30, such as go beyond the scope, with dedicated buffering liquid or 0.872mol/L
The adjustment of NaHCO3 liquid.
(5) filters pressing separates, and obtains III supernatant F III of F precipitating, and III supernatant of F is continued separation component II, detected using densitometer
III supernatant density of F is 0.9794 ~ 0.9746 g/cm3, uses biuret method detection protein content for 3.8 ~ 5.6g/L, FIII
Sediment weight scope control is throwing slurry amount × (0.07419 ± 3 × 0.00267).
Using production method of the invention, following income can be obtained:
It using the present invention of technical solution as above, has the advantages that compared with the existing technology: F II can be ensured using this law
+ III precipitating in protein can dissolve out completely, in process of production by control liquid feeding speed, solution temperature, liquid feeding mode,
The modes such as pH and concentration of alcohol of control, it is ensured that intravenous human immunoglobulin(HIg) (pH4) purity, molecular size distribution, PKA, Fc biology
The indexs such as activity, anti-complement activity, which meet the requirements, to be shown in Table: 1.This law to the filters pressing separation density of supernatant, protein content and
Component III Sediment weight quality control, accurately control in supernatant influence cold ethanol separation protein content, pH, temperature,
Five factors of concentration of alcohol and ionic strength.
Dedicated buffering liquid (4.0mol/L acetic acid, 0.8mol/L sodium acetate).
Table 1 intravenous human immunoglobulin(HIg) (pH4) quality index situation
| Purity (%) | Molecular size is distributed (%) | PKA(IU/ml) | Fc biological activity (%) | Anti-complement activity (%) | |
| Chinese Pharmacopoeia standard | ≥95.0 | ≥95.0 | ≤35 | Nothing | ≤50 |
| Average value ± SD | 99.0±0.2 | 98.9±0.5 | ≤1 | 99.6±13 | 17±5 |
(1) dissolution that F II+III is precipitated: II+III theoretical weight (A) of F for not adding diatomite is calculated, formula: A=plasma volume (kg) ×
0.04。
Calculate the amount that dissolution F II+III precipitates water for injection used, formula: injection water (kg)=blood plasma needed for dissolving
It measures (kg) × 0.6405-F II+III and precipitates (kg).Dissolving F II+III to precipitate the temperature of water for injection used is 0 ~ 4 DEG C, controls F
II+III precipitating lysate temperature is 0 ~ 4.0 DEG C.It precipitates within stirring 2 ~ 4 hours and is completely dissolved to F II+III.
(2) sodium acetate solution is added: calculating the amount that sodium acetate solution is added, formula: NaAc 3H2O (kg)=constant A
×0.054；Injection water (kg)=constant A × 2；Sodium acetate solution amount (kg)=plasma volume (kg) × 0.08216, acetic acid
Sodium solution temperature is controlled at 0 ~ 4.0 DEG C.By sodium acetate solution, constant speed is added in II+III lysate of F in 4 ~ 5 hours, continues to stir
It mixes 1 ~ 2 hour.
(3) buffer is added and adjusts pH: calculating the amount that dedicated buffering liquid is added, formula: dedicated buffering liquid additional amount (kg)
=(II+III+0.24 × plasma volume of precipitation capacity kg (kg) of F+sodium acetate solution amount (kg)) × 0.0043, the dilution of dedicated buffering liquid
With injection water (kg)=constant A, both dedicated buffering liquid and water for injection are uniformly mixed, control mixeding liquid temperature to 0 ~
4.0℃.By dedicated buffering liquid and water for injection mixed liquor, constant speed is added in reactor tank in 2 ~ 3 hours, is adjusted in reactor tank and is hanged
Liquid pH value is to 5.15 ~ 5.25.
(4) be added ethyl alcohol: calculate be added amount of alcohol: formula: be added amount of alcohol (kg)=reactor tank in suspension total amount (kg) ×
0.180.Cool down to product, temperature is down to 0 ~ 1.0 DEG C in one hour.By 95% ethyl alcohol (- 15.0 DEG C or less) at 3 ~ 5 hours
Interior constant speed is added in reactor tank, until the ethyl alcohol of suspension final concentration of 17%, temperature is down to -5.0 ~ -6.0 DEG C.It finishes and continues to stir
It samples within 10 minutes, surveys suspension pH value, should be 5.10 ~ 5.30, such as go beyond the scope, with dedicated buffering liquid or 0.872mol/L
NaHCO3Liquid adjustment.
(5) filters pressing separates, and obtains III supernatant F III of F precipitating, and III supernatant of F is continued separation component II, detected using densitometer
III supernatant density of F is 0.9794 ~ 0.9746 g/cm3, use biuret method detection protein content for 3.8 ~ 5.6g/L, FIII
Sediment weight scope control is throwing slurry amount × (0.07419 ± 3 × 0.00267) kg.
It using the present invention of technical solution as above, has the advantages that compared with the existing technology: can be true using this law
The protein protected in the precipitating of F II+III can dissolve out completely, and it is abundant to solve components precipitate protein in plasma protein separation process
The problem of dissolution.
On basic principles and main features and advantages of the present invention of the invention have been shown and described.Those skilled in the art
Member is it should be recognized that the present invention is not limited to the above embodiments, and the above embodiments and description only describe the present invention
Principle, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these variation and
Improvement is both fallen in claimed range.
Claims (7)
1. a kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III, which is characterized in that include following step
It is rapid:
(1) II+III precipitate production method of F: refrigerated plasma with dedicated buffering liquid adjusts blood plasma pH value to 7.15 ~ 7.25 after melting,
Dedicated buffering liquid is acetic acid/sodium acetate solution of pH4.0；Add -15 DEG C of 95% ethyl alcohol below into reactor tank, until second in tank
Alcohol final concentration of 8%, blood plasma temperature are controlled at -1.5 ~ -2.5 DEG C；Filters pressing separation obtains component I, i.e. FI precipitating and FI supernatant；To
The mixed liquor of dedicated buffering liquid and ethyl alcohol is added in FI supernatant, makes ethyl alcohol final concentration of 22%, -5.0 ~ -6.0 DEG C of liquid temperature, continues
Stirring 2.5 hours, surveying pH value is 6.70 ~ 6.80, otherwise with dedicated buffering liquid or the NaHCO of 0.872mol/L3Adjustment, addition help
Filtering agent perlite, diatomite are separated after stirring 45 minutes with filter press, collect component, which is divided into the precipitating of F II+III；
(2) control dissolution F II+III precipitates dissolution water and lysate temperature is 0 ~ 4.0 DEG C, and mixing time is 2 ~ 4 hours；
(3) control of sodium acetate solution temperature is added at 0 ~ 4.0 DEG C；The timing since be initially added into sodium acetate solution, time control
In 4 ~ 5 hours, mixing time control was at 1 ~ 2 hour after addition；
(4) when adjusting pH, uniformly being mixed with both dedicated buffering liquid and water for injection for pH will be adjusted and be added, mixed liquor is controlled
Temperature is to 0 ~ 4.0 DEG C；The liquid feeding time controlled in 2 ~ 3 hours, and suspension pH value is controlled 5.15 ~ 5.25；
(5) it is added before ethyl alcohol and lysate cooling is precipitated to F II+III, temperature is down to 0 ~ 1.0 DEG C in one hour；
(6) be added concentration of alcohol be 95%, temperature control at -15.0 DEG C hereinafter, from be added ethyl alcohol timing, from addition second
Alcohol starts timing, and the liquid feeding time controlled at 3 ~ 5 hours, and lysate temperature is down to -5.0 ~ -6.0 by ethyl alcohol final concentration of 17%
℃；Suspension pH value should be 5.10 ~ 5.30 after ethyl alcohol is added；
(7) filter aid diatomite is added to be separated after stirring 15 minutes with filter press, obtains component FII) it precipitates and FIII supernatant；
(8) densimeter method detection III supernatant density of F is 0.9794 ~ 0.9746 g/cm3, biuret method detection protein content be
3.8 ~ 5.6g/L, FIII Sediment weight scope control are throwing slurry amount × (0.07419 ± 3 × 0.00267) kg.
2. a kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III as described in claim 1,
It is characterized in that, the production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III are as follows:
The dissolution that F II+III is precipitated in step (2): II+III theoretical weight (A) of F for not adding diatomite, formula: A=blood plasma are calculated
It measures (kg) × 0.04；The amount that dissolution F II+III precipitates water for injection used is calculated, formula: injection water (kg) needed for dissolving=
Plasma volume (kg) × 0.6405-F II+III precipitates (kg)；Dissolving F II+III to precipitate the temperature of water for injection used is 0 ~ 4 DEG C, control
It is 0 ~ 4.0 DEG C that F II+III processed, which precipitates lysate temperature,；It precipitates within stirring 2 ~ 4 hours and is completely dissolved to F II+III.
3. a kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III as described in claim 1,
It is characterized in that, sodium acetate solution is added in step (3): calculating the amount that sodium acetate solution is added, formula: NaAc 3H2O(kg)
= A×0.054；Injection water (kg)=constant A × 2；Sodium acetate solution amount (kg)=plasma volume (kg) × 0.08216, second
Acid sodium solution temperature is controlled at 0 ~ 4.0 DEG C；By sodium acetate solution, constant speed is added in II+III lysate of F in 4 ~ 5 hours, is continued
Stirring 1 ~ 2 hour.
4. a kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III as described in claim 1,
It is characterized in that, buffer is added in step (4) and adjusts pH: calculating the amount that dedicated buffering liquid is added, formula: dedicated buffering liquid adds
Enter amount (kg)=(F II+III precipitation capacity (kg)+0.24 × plasma volume (kg)+sodium acetate solution amount (kg)) × 0.0043, it is dedicated
Buffer dilution injection water (kg)=constant A uniformly mixes both dedicated buffering liquid and water for injection, control mixing
Liquid temperature is to 0 ~ 4.0 DEG C；By dedicated buffering liquid and water for injection mixed liquor, constant speed is added in reactor tank in 2 ~ 3 hours, adjustment
Suspension pH value is to 5.15 ~ 5.25 in reactor tank.
5. a kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III as described in claim 1,
It is characterized in that, ethyl alcohol is added in step (6): calculating and amount of alcohol: formula is added: suspension in amount of alcohol (kg)=reactor tank is added
Total amount (kg) × 0.180；Cool down to product, temperature is down to 0 ~ 1.0 DEG C in one hour；By 95% ethyl alcohol (- 15.0 DEG C or less)
Constant speed is added in reactor tank in 3 ~ 5 hours, until the ethyl alcohol of suspension final concentration of 17%, temperature is down to -5.0 ~ -6.0 DEG C；It finishes
Continue to sample for stirring 10 minutes, survey suspension pH value, should be 5.10 ~ 5.30, such as go beyond the scope, with dedicated buffering liquid or
The adjustment of 0.872mol/L NaHCO3 liquid.
6. a kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III as described in claim 1,
It is characterized in that, filters pressing separates in step (7) a metallic, obtains III supernatant F III of F precipitating, and III supernatant of F continues separation component II, uses
It is 0.9794 ~ 0.9746 g/cm that densitometer, which detects III supernatant density of F,3, use biuret method detection protein content for 3.8 ~
5.6g/L, FIII Sediment weight scope control are throwing slurry amount × (0.07419 ± 3 × 0.00267) kg.
7. a kind of production method of intravenous human immunoglobulin(HIg) (pH4) cold ethanol component III as described in claim 1,
It is characterized in that, the product variety that this technique is suitable for production includes: intravenous human immunoglobulin(HIg) (pH4) liquid and freeze-dried formulation
Each specification [0.5g/ bottles (5 %, 10ml), lg/ bottles (5 %, 20ml), 1.25g/ bottles (5 %, 25ml), 2. 5 g/
Bottle (5 %, 50ml), 5g/ bottles (5%, 100ml), 10g/ bottles (5%, 200ml)] and human immunoglobulin(HIg) class product.
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Cited By (4)
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| CN112378899A (en) * | 2020-10-27 | 2021-02-19 | 西北矿冶研究院 | Method for determining gold in crude lead sample by low-temperature separation ICP-AES (inductively coupled plasma-atomic emission Spectrometry) method |
| 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 |
| CN116731162A (en) * | 2023-06-09 | 2023-09-12 | 广东丹霞生物制药有限公司 | Human immunoglobulin production process |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112378899A (en) * | 2020-10-27 | 2021-02-19 | 西北矿冶研究院 | Method for determining gold in crude lead sample by low-temperature separation ICP-AES (inductively coupled plasma-atomic emission Spectrometry) method |
| 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 |
| CN116731162A (en) * | 2023-06-09 | 2023-09-12 | 广东丹霞生物制药有限公司 | Human immunoglobulin production process |
| CN116731162B (en) * | 2023-06-09 | 2024-03-19 | 广东丹霞生物制药有限公司 | Human immunoglobulin production process |
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