CN100569287C - Dog immunoglobulin'intravenous, its preparation method and dog intravenous immune globulin preparation - Google Patents
Dog immunoglobulin'intravenous, its preparation method and dog intravenous immune globulin preparation Download PDFInfo
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- CN100569287C CN100569287C CNB2004100395712A CN200410039571A CN100569287C CN 100569287 C CN100569287 C CN 100569287C CN B2004100395712 A CNB2004100395712 A CN B2004100395712A CN 200410039571 A CN200410039571 A CN 200410039571A CN 100569287 C CN100569287 C CN 100569287C
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Abstract
The present invention relates to dog immunoglobulin'intravenous, its preparation method and preparation thereof; be by blood plasma or serum from healthy dogs; the pH value that separates, prepares in conjunction with cold ethanol method through cold ethanol method or ethacridine is 3.0～5.4; purity be not less than 95% can be used for the intravenous immunoglobulin of dog; and being mixed with one or both protective agents that contain in 5～15% maltose, 5～10% glucoses or 3～10% glucose salines with this immunoglobulin, protein content is 1～15% Imnunoglobulin injection simultaneously.
Description
Technical field
The present invention relates to biological technical field, be specifically related to a kind of dog immunoglobulin'intravenous, its preparation method and dog intravenous immune globulin preparation.
Background of invention
The status of house pet in people's daily life is more and more important, and pet care has become the problem that a worker wants.Dog is that separation and Extraction gets biological product from dog plasma with blood products, claims blood plasma derivant again.This based article is used for the treatment or the passive immunity of dog disease.
One of above-mentioned biological product are immunoglobulin (Ig), and it is the important component part of body immune system, are bringing into play very important effect in defence infection and immunomodulating.
On the use approach, immunoglobulin and immunoglobulin preparation generally are divided into intramuscular injection immunoglobulin (IMIG) and immunoglobulin'intravenous (IVIG) two classes, now existing inventor studies with immunoglobulin dog and prepares, as patent application 00123292.4, but, owing to technical reason, it can only develop the immunoglobulin (IMIG) of oral or intramuscular injection, has significant limitation in the use, mainly show: (1) consumption is too small, even the injection of multi-section position is Zong consumption also has only 20 milliliters; (2) injection site has the serious pain reaction; (3) have part Ig by enzymatic degradation before entering circulation, and onset is slower.The mechanism of action of immunoglobulin'intravenous (IVIG) is passive immunity, be mainly used to prevention and treat viral, bacterial infection, and various immunodeficiency diseases and immunocompromised disease etc., be mainly reflected in that antibody replenishes and immunomodulating, comprise target cell blocking effect, immune opsonic action, in conjunction with activatory complement component, change immune complex structure and solubility, improve IgG the metabolism rate, increase the quantity of natural killer cell in the tip blood and strengthen its function, in and autoimmune antibody and the expression etc. of regulating the various kinds of cell factor.
At present, still dog is not used the research of used for intravenous injection immunoglobulin and the bibliographical information of application.
Summary of the invention
The object of the present invention is to provide a kind of dog immunoglobulin'intravenous.
Another object of the present invention is to provide the preparation method of dog immunoglobulin'intravenous.
The present invention also is to provide a kind of dog intravenous immune globulin preparation with purpose.
At first, a kind of dog immunoglobulin'intravenous provided by the invention, be by blood plasma or serum from healthy dogs, the pH value that separates, prepares in conjunction with cold ethanol method through cold ethanol method or ethacridine is 3.0～5.4, purity be not less than 95% can be used for the intravenous immunoglobulin of dog.
In the above-mentioned dog immunoglobulin'intravenous, described blood plasma or serum comprise pooled plasma or the serum gathered after a large amount of healthy dogs of the para-influenzal vaccine immunity of parvovirus infection, canine distemper, infectious canine hepatitis, rabies and dog from multiple dog disease.
In the above-mentioned dog immunoglobulin'intravenous, described cold ethanol method comprises following steps:
1). blood plasma or serum are melted, be cooled to 0 ℃ gradually, control pH7.0～7.4 add pre-cooled ethanol to final concentration 8% (g/100ml), centrifugal removal foreign protein;
2). repeatedly supernatant is handled, adjusted pH4.5～6.0, add ethanol gradually to final concentration 15%～20%, after the standing over night, centrifugal reservation precipitation;
3), above-mentioned precipitation adds 10 times of amounts of 0 ℃ of no heat source water, 0.5mol/LNa
2HPO
4Adjust pH4.5～5.0 with 0.5mol/L acetic acid, stirred standing over night 3 hours.
4), above-mentioned lysate is centrifugal with continuous centrifuge, second-3 plate filters supernatant, 0.5mol/L Na
2HPO
4Transfer pH4.8～5.3 with 0.5mol/L acetic acid, add ethanol to 15%～20%, stirred standing over night 1 hour.
5), the supernatant behind the centrifugalize foreign protein transfers to pH6.5～7.0, adds ethanol to 22～26%, standing over night obtains immunoglobulin suspension, centrifugation obtains immunoglobulin;
6). the immunoglobulin that step 5) is obtained carries out post processing in pH3.5～5.0, and ultrafiltration, degerming, packing to obtain can intravenous protein content be 1～15% immunoglobulin.
In the above-mentioned dog immunoglobulin'intravenous, described ethacridine comprises following steps in conjunction with cold ethanol method:
1). will melt back blood plasma and serum and transfer to pH 8.0～9.0, add 2% (g/100ml) ethacridine, remove the albumin precipitation, get supernatant;
2). with supernatant activated carbon adsorption after-filtration, under 0 ℃, pH6.5～7.0 conditions, add ethanol to 22～26%, standing over night obtains immunoglobulin suspension, and centrifugation obtains immunoglobulin;
3). with step 2) immunoglobulin that obtains carries out post processing in pH3.0～5.4, and ultrafiltration, degerming, packing to obtain intravenous protein content to be the Imnunoglobulin injection of 1～15% (g/100ml).
In the above-mentioned dog immunoglobulin'intravenous; described post processing is for adding protective agent in the immunoglobulin solution of pH3.0～5.4, described protective agent is one or both the combination in the saline of maltose, 5～10% glucoses or 3～10% glucoses of 5～15% (g/100ml).
The present invention provides a kind of preparation method of dog immunoglobulin'intravenous simultaneously, comprises the steps:
1). remove albumin: the blood plasma of healthy dogs or serum in pH4.8～5.1 o'clock, are added ethanol to 15～20%, remove foreign protein; Or use rivanol method, and will melt back blood plasma and serum and transfer to pH 8.0～9.0, add 2% ethacridine, remove albumin;
2). extract immunoglobulin: will remove solution behind foreign protein or the albumin under 0 ℃, pH6.5～7.0 conditions, adding ethanol to 22～26%, standing over night obtains immunoglobulin suspension, and centrifugation obtains immunoglobulin;
3). preparation Imnunoglobulin injection: with step 2) immunoglobulin that obtains carries out post processing in pH3.0～5.4, and ultrafiltration, degerming, packing to obtain can intravenous protein content be 1～15% Imnunoglobulin injection.
In the preparation method of above-mentioned dog immunoglobulin'intravenous; the post processing of step 3) is for adding protective agent in the immunoglobulin solution of pH3.0～5.4, described protective agent is one or both the combination in the saline of 5～15% maltose, 5～10% glucoses or 5～10% glucoses.
In the preparation method of above-mentioned dog immunoglobulin'intravenous, the blood plasma of healthy dogs or serum are taken from multiple dog disease in the step 1), comprise pooled plasma or the serum gathered after a large amount of healthy dogs of the para-influenzal vaccine immunity of parvovirus infection, canine distemper, infectious canine hepatitis, rabies and dog.
The present invention also provides the dog intravenous immune globulin preparation, by blood plasma or serum from healthy dogs, the pH value that separates, prepares in conjunction with cold ethanol method through cold ethanol method or ethacridine is 3.0～5.4, and purity is not less than 95% the intravenous immunoglobulin of dog that can be used for, and to be prepared into protein content be 1～10% Imnunoglobulin injection.
Above-mentioned dog intravenous immune globulin preparation also comprises the acceptable protective agent of organism, and described protective agent is one or both the combination in the saline of 5～15% maltose, 5～10% glucoses or 3～10% glucoses.
Dog immunoglobulin'intravenous provided by the invention, it maintains complete natural biological molecular structure, has whole physiologically actives of immunoglobulin G (IgG) in the body, immunoglobulin IgG, IgG subclass constituent ratio is similar to normal dogs blood plasma.
The preparation method of dog immunoglobulin'intravenous provided by the invention, technology is simple, and cost is low, response rate height, the finished product purity of preparation is good, and purity surpasses 96%, and stable performance.
Dog intravenous immune globulin preparation provided by the invention does not contain any antiseptic, and biocompatibility is good.
Description of drawings
Fig. 1 is the immunoglobulin molecules amount evaluation figure that the inventive method obtains.
The specific embodiment
For opener, below from several respects narration the present invention.
Dog immunoglobulin'intravenous provided by the invention (IVIG) is with blood plasma or the serum of dog, uses the modern biotechnology separation and Extraction, through inactivation of virus and add the dog that an amount of stabilizing agent makes and use the intravenous injection biological preparation.Wherein, blood plasma and serum can be the pooled plasma that obtains after a large amount of healthy dogs of vaccine immunity of multiple dog disease (as parvovirus infection, canine distemper, infectious canine hepatitis, rabies, dog parainfluenza etc.), at this moment, to the immunity of dog, get the blood plasma process, identical with other conventional way; The method of inactivation of virus and step can adopt conventional method; Modern bioseparation technology is meant the method for separation and Extraction immunoglobulin from blood plasma in the present invention, can for cold ethanol method or ethacridine in conjunction with cold ethanol method, the general requirement during extraction is:
1, raw material
The liquid blood plasma of fresh separated, serum, slight haemolysis or fatty blood plasma, the component etc. of removing other plasma proteins all can be used for preparation.
2, ingredient requirement
Used blood plasma or serum should keep aseptic as far as possible, make or the cryogenic freezing preservation otherwise should in time feed intake.Cryogenic freezing is preserved the long storage life of blood plasma should be above 2 years.
3, operationlocation
The operating room should meet GMP technological process requirement.Freezer and various production apparatus must be special-purpose, forbid to use with other foreign protein matter.The building of operating room should be convenient to cleaning, sterilization, mildew-resistant.In manufacture process,, should take various effective measures,, note sterile working etc. as reducing the operating room temperature for preventing that goods from polluting pyrogen.The apparatus of various direct contact goods, with after should clean immediately, with before must be through heat extraction protoplasm or sterilization treatment.
4, preparation water and chemical drugs
Industrial water should meet drinking water standard, and the water that is directly used in goods should meet the water for injection standard.Used various chemical drugs should meet regulations such as " Chinese veterinary drug allusion quotation ", " People's Republic of China's veterinary biologics quality standard ", " Chinese biological goods main raw material(s) tentative standard ", does not include standard person in and should be not less than chemical pure.
In technical solution of the present invention and following examples, if no special instructions, mentioned material concentration such as immunoglobulin concentration, concentration of alcohol and glucose, maltose, glucosamine salt water concentration etc. are all represented with %, mean g/100ml, the weight in grams of contained solute in promptly per 100 ml solns.
Embodiment one: adopt cold ethanol method to extract the dog immunoglobulin'intravenous
Described cold ethanol method comprises following steps:
1). melt slurry and add ethanol
The blood plasma that will meet ingredient requirement melts under 37 ℃ of conditions for 200,000 milliliters fast, and inserts retort under gnotobasis; Being cooled to 0 ℃ under stirring gradually, is 4.0 acetate buffer solution accent pH to 7.2 with pH; Adding pre-cooling while stirring and be-15 ℃--20 ℃ 53.3% ethanol to ethanol volume final concentration is that 8% (add ethanol and want slowly, in 100,000 milliliters of blood plasma, per minute adds 400-500ml, added in 2-3 hour), temperature maintenance is at-2 ℃--and 3 ℃, add the back and continue to stir 1 hour, left standstill 2 hours.
2). remove foreign protein
The use continuous centrifuge is centrifugal, centrifuge liquid outlet temperature-2 ℃～-3 ℃, and supernatant pH is that to transfer pH be 5.8 for 4.0 acetate buffer solution, it is 19% that cold ethanol with 95% slowly adds to final volume concentration, temperature remains on-5 ℃～-6 ℃, adds the back and stirs-5 ℃ of standing over night 2 hours; For the second time centrifugal, keep precipitation.
Above-mentioned precipitation adds 10 times of amounts of 0 ℃ of no heat source water, 0.5mol/L Na
2HPO
4Adjust pH4.8 with 0.5mol/L acetic acid, stirred standing over night 3 hours; Lysate is centrifugal with continuous centrifuge, and second-3 plate filters supernatant, 0.5mol/L Na
2HPO
4Transfer pH5.1 with 0.5mol/L acetic acid, add ethanol to 17%, stirred standing over night 1 hour.
3), separating immune globulin
Supernatant behind the centrifugal removal foreign protein is adjusted pH6.9, adds ethanol to 25%, and standing over night obtains immunoglobulin suspension, and centrifugation obtains immunoglobulin.
4). ultrafiltration and concentration
The immunoglobulin that step 3) is obtained carries out post processing at pH4.0, and ultrafiltration and concentration to obtain can intravenous protein content be 1～15% immunoglobulin.
Embodiment two: adopt ethacridine to extract the dog immunoglobulin'intravenous in conjunction with cold ethanol method
1) melts slurry and accent pH
The blood plasma that will meet ingredient requirement melts under 37 ℃ of conditions for 200,000 milliliters fast, and inserts retort under gnotobasis; Slow adding concentration is 1% Na under stirring
2CO
3, transfer pH to 8.5 ± 0.1.
2). remove albumin
Get dog plasma and add retort, under stirring, in retort, add and isopyknic 2% ethacridine of blood plasma, stop after adding stirring, left standstill 2 hours, get supernatant and add 2% active carbon, stirred 30 minutes, left standstill 1 hour, cross second-3 plate, collect filtrate and spend the night.
3). the precipitation immunoglobulin
With step 2) filtrate be cooled to 0 ℃, add 25% NaCl solution to final concentration 1%, 0.5mol/L acetate buffer and transfer pH to 6.9, add 95% ethanol to final concentration 25%, stirred 1 hour, after the standing over night, the solution continuous centrifugal, precipitation is the dog immunoglobulin.
4). ultrafiltration and concentration
The immunoglobulin that step 3) is obtained carries out post processing at pH4.0, and ultrafiltration and concentration to obtain can intravenous protein content be 1～15% immunoglobulin.
Embodiment three: preparation dog intravenous immune globulin preparation-maltose injection
On the basis of embodiment one or embodiment two; step below continuing: in embodiment three step 4),, add maltose to 10% with solution concentration; degerming packing then obtains protein content and is 5% the protectant dog immunoglobulin'intravenous of interpolation maltose injection.
Use the same method, change the addition of maltose, make any desired concentration change of content between 5% to 10% of maltose to obtain immunoglobulin content between 1% to 15% that maltose concentration is at 5% to 10% dog immunoglobulin'intravenous injection.
Embodiment four: preparation dog intravenous immune globulin preparation-glucose injection
The method same with embodiment three replaces maltose to make protective agent with glucose, obtains immunoglobulin content between 1% to 15%, and concentration of glucose is at 5% to 10% dog immunoglobulin'intravenous injection.
Embodiment five: preparation dog intravenous immune globulin preparation-glucosamine salt water injection
The method same with embodiment three replaces maltose to make protective agent with glucose saline, obtains immunoglobulin content between 1% to 15%, and the glucosamine salt water concentration is at 5% to 10% dog immunoglobulin'intravenous injection.
Embodiment six: preparation dog intravenous immune globulin preparation-maltose glucose injection
The method same with embodiment three share maltose and glucose is made protective agent, obtains immunoglobulin content between 1% to 15%, maltose concentration 5% to 10%, concentration of glucose is at 5% to 10% dog immunoglobulin'intravenous injection.
Embodiment seven: preparation dog intravenous immune globulin preparation-maltose glucosamine salt water injection
The method same with embodiment three; share maltose and glucose saline is made protective agent; obtain immunoglobulin content between 1% to 15%, maltose concentration 5% to 10%, concentration of glucose is at 5% to 10% brinish dog immunoglobulin'intravenous injection.
The above-mentioned IVIG that obtains is carried out performance test:
1, places test
After the liquid preparation packing, placed under 20～35 ℃ condition at least 30 days, check by bottle, outward appearance does not contain any foreign body for slightly thickness, yellow or green brown clear liquid extremely, does not have muddiness and deposited phenomenon yet.
2, heat stabilization test
Get liquid preparation inspection product, put into 57 ± 0.5 ℃ of water bath heat preservations after 4 hours, gel-free forms or floccule occurs.
3, chemical assay
Undertaken by " People's Republic of China's veterinary biologics quality standard ", " biological product chemical assay rules " etc.:
3.1 pH value
1% Imnunoglobulin injection is 20 ± 2 ℃ of mensuration, and its pH value is 6.4～7.4; 5% and 10% goods are diluted to 1% protein concentration with normal saline, and 20 ± 2 ℃ of mensuration, pH value is 6.4～7.4.
3.2 protein content and total amount
Measure with the wolframic acid sedimentation method, protein content is not less than 95% of labelled amount, and every bottle of total protein is not less than the labelled amount of product specification.
3.3 purity
The goods of different size, immunoglobulin content are not less than 95% of total protein.
3.4 sugared content
If sugaring (glucose, maltose etc.) in the goods, its content should not be higher than 50g/L.
3.5 molecular size distribution
IgG monomer and dimer content sum are not less than 90.0%.
4, discrimination test
Adopt immune double diffusion method.Only produce precipitation line, do not produce precipitation line with the serum of anti-people, anti-horse, anti-cattle with the serum of anti-dog.
5, sterility test
Undertaken by " People's Republic of China's veterinary biologics quality standard ".
Result: conformance with standard.
6, safety test
6.1 Cavia porcellus test
With 2 of body weight 300～400g healthy guinea pigs, every lumbar injection inspection product 5ml, the injection back in half an hour animal do not have tangible abnormal response, observed 7 days, animal all is good for and deposits, it is qualified that every weight increase person is judged to.As do not meet above-mentioned requirements, with 4 Cavia porcellus retrials once, criterion is the same.
Result: conformance with standard.
6.2 white mice test
With 5 of body weight 18～20g white mice, every lumbar injection inspection product 0.5ml, animal does not have tangible abnormal response in half an hour, continues to observe 7 days, and animal all is good for and deposits, and it is qualified that every weight increase person is judged to.As do not meet above-mentioned requirements, with 10 white mice retrials once, criterion is the same.
Result: conformance with standard.
7, pyrogen test
Undertaken by " People's Republic of China's veterinary biologics quality standard ", " biological product chemical assay rules " etc.Injected dose is by rabbit body weight injection 0.6g/kg.
Result: conformance with standard.
Above-mentioned dog immunoglobulin'intravenous that obtains and dog intravenous immune globulin preparation can be applicable to prevention and treat viral, bacterial infection, and various immunodeficiency disease and immunocompromised disease etc.
Dog immunoglobulin'intravenous provided by the invention and dog intravenous immune globulin preparation are colourless clear liquid, its using method:
Venoclysis.If when needing heavy dose of infusion, can number bottle product continuous infusions (must in 4 hours, use).Carry out intravenous drip again after glucose injection dilution this product of available in case of necessity 5%.
Infusion dosage: dog below 5 kilograms, 5 milliliters/day, logotype 2 ~ 3 days; 5 ~ 10 kilograms of dogs, 10 milliliters/day, logotype 2 ~ 3 days; Dog more than 10 kilograms, 10 ~ 20 milliliters/day, logotype 2 ~ 3 days.When being in a bad way, can take the circumstances into consideration to double.
Infusion velocity: use first, begin in 30 minutes infusion velocity and is 0.02 ~ 0.03 milliliter/kg body weight/minute, as if not seeing untoward reaction, rise to 0.04 ~ 0.08 milliliter/kg body weight/minute.
The dog of 1:3 kilogram for example, the first speed of annotating 30 minutes is 3～5 droplets/minute; Speed is 8～15 droplets/minute after 30 minutes.
The dog of 2:10 kilogram for example, first notes speed is 8～15; Speed is 16～25 droplets/minute after 30 minutes.
Preserve and effect duration: be stored in 2～8 ℃ of dark places, guard against and freeze; 3 years effect duration.
Claims (8)
1. dog immunoglobulin'intravenous, be by blood plasma or serum from healthy dogs, the pH value that separates, prepares in conjunction with cold ethanol method through cold ethanol method or ethacridine is 3.0～5.4, purity be not less than 95% can be used for the intravenous immunoglobulin of dog;
Described cold ethanol method comprises following steps:
1). blood plasma or serum are melted, be cooled to 0 ℃ gradually, control pH7.0～7.4, adding pre-cooled ethanol to final concentration 8% is 8g/100ml, centrifugal removal foreign protein;
2). repeatedly supernatant is handled, promptly adjusted pH4.5～6.0, add ethanol gradually to final concentration 15%～20%, after the standing over night, centrifugal reservation precipitation;
3), above-mentioned precipitation adds 10 times of amounts of 0 ℃ of no heat source water, 0.5mol/L Na
2HPO
4Adjust pH4.5～5.0 with 0.5mol/L acetic acid, stirred standing over night 3 hours;
4), the solution of step 3) is centrifugal with continuous centrifuge, second-3 plate filters supernatant, 0.5mol/L Na
2HPO
4Transfer pH4.8～5.3 with 0.5mol/L acetic acid, add ethanol to 15%～20%, stirred standing over night 1 hour;
5), the supernatant behind the centrifugalize foreign protein transfers to pH6.5～7.0, adds ethanol to 22～26%, standing over night obtains immunoglobulin suspension, centrifugation obtains immunoglobulin;
6). the immunoglobulin that step 5) is obtained carries out post processing in pH3.5～5.0, and ultrafiltration, degerming, packing to obtain can intravenous protein content be 1～15% immunoglobulin;
Described ethacridine comprises following steps in conjunction with cold ethanol method:
1). will melt back blood plasma or serum and transfer to pH 8.0～9.0, adding 2% is the 2g/100ml ethacridine, removes the albumin precipitation, gets supernatant;
2). with supernatant activated carbon adsorption after-filtration, under 0 ℃, pH6.5～7.0 conditions, add ethanol to 22～26%, standing over night obtains immunoglobulin suspension, and centrifugation obtains immunoglobulin;
3). with step 2) immunoglobulin that obtains carries out post processing in pH3.0～5.4, and ultrafiltration, degerming, packing to obtain can intravenous protein content be 1～15% i.e. Imnunoglobulin injection of 1～15g/100ml.
2. dog immunoglobulin'intravenous as claimed in claim 1; it is characterized in that: described post processing is for adding protective agent in the immunoglobulin solution of pH3.0～5.4, and described protective agent is 5～15% i.e. combinations of one or both in maltose, 5～10% glucoses and 3～10% glucose salines of 5～15g/100ml.
3, a kind of preparation method of dog immunoglobulin'intravenous comprises the steps:
1). blood plasma or serum are melted, be cooled to 0 ℃ gradually, control pH7.0～7.4, adding pre-cooled ethanol to final concentration 8% is 8g/100ml, centrifugal removal foreign protein;
2). repeatedly supernatant is handled, promptly adjusted pH4.5～6.0, add ethanol gradually to final concentration 15%～20%, after the standing over night, centrifugal reservation precipitation;
3), above-mentioned precipitation adds 10 times of amounts of 0 ℃ of no heat source water, 0.5mol/L Na
2HPO
4Adjust pH4.5～5.0 with 0.5mol/L acetic acid, stirred standing over night 3 hours;
4), step 3) solution is centrifugal with continuous centrifuge, second-3 plate filters supernatant, 0.5mol/L Na
2HPO
4Transfer pH4.8～5.3 with 0.5mol/L acetic acid, add ethanol to 15%～20%, stirred standing over night 1 hour;
5), the supernatant behind the centrifugalize foreign protein transfers to pH6.5～7.0, adds ethanol to 22～26%, standing over night obtains immunoglobulin suspension, centrifugation obtains immunoglobulin;
6). the immunoglobulin that step 5) is obtained carries out post processing in pH3.5～5.0, and ultrafiltration, degerming, packing to obtain can intravenous protein content be 1～15% immunoglobulin.
4. the preparation method of dog immunoglobulin'intravenous as claimed in claim 3; it is characterized in that: the post processing of step 3) is for adding protective agent in immunoglobulin solution, and described protective agent is 5～15% i.e. combinations of one or both in maltose, 5～10% glucoses and 5～10% glucose salines of 5～15g/100ml.
5, a kind of preparation method of dog immunoglobulin'intravenous comprises the steps:
1). will melt back blood plasma or serum and transfer to pH 8.0～9.0, adding 2% is the 2g/100ml ethacridine, removes the albumin precipitation, gets supernatant;
2). with supernatant activated carbon adsorption after-filtration, under 0 ℃, pH6.5～7.0 conditions, add ethanol to 22～26%, standing over night obtains immunoglobulin suspension, and centrifugation obtains immunoglobulin;
3). with step 2) immunoglobulin that obtains carries out post processing in pH3.0～5.4, and ultrafiltration, degerming, packing to obtain can intravenous protein content be 1～15% i.e. Imnunoglobulin injection of 1～15g/100ml.
6. the preparation method of dog immunoglobulin'intravenous as claimed in claim 5; it is characterized in that: the post processing of step 3) is for adding protective agent in immunoglobulin solution, and described protective agent is 5～15% i.e. combinations of one or both in maltose, 5～10% glucoses and 5～10% glucose salines of 5～15g/100ml.
7. dog intravenous immune globulin preparation, being prepared into immunoglobulin content by the described immunoglobulin of claim 1 is 1～15% Imnunoglobulin injection.
8. dog intravenous immune globulin preparation according to claim 7; it is characterized in that; also comprise the acceptable protective agent of organism, described protective agent is 5～15% i.e. combinations of one or both in the saline of maltose, 5～10% glucoses or 3～10% glucoses of 5～15g/100ml.
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| US7682619B2 (en) | 2006-04-06 | 2010-03-23 | Cornell Research Foundation, Inc. | Canine influenza virus |
| CN102949719A (en) * | 2012-04-28 | 2013-03-06 | 上海市徐汇区中心医院 | Application of intravenous immunoglobulins (IVIg) in inhibiting cholera toxin and galectin from being combined into ganglioside GM1 |
| CN104784689A (en) * | 2015-05-05 | 2015-07-22 | 王宏伟 | Canine parvovirus specific immune globulin preparation as well as preparation method and application thereof |
| CN109793894A (en) * | 2019-04-03 | 2019-05-24 | 长春西诺生物科技有限公司 | Application of the dog immunoglobulin in preparation dog hemolytic anemia drug |
| CN110590936B (en) * | 2019-10-16 | 2021-03-16 | 长春西诺生物科技有限公司 | Purification method of albumin in dog serum |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1042716A (en) * | 1988-11-17 | 1990-06-06 | 中国医学科学院输血研究所 | Rivanol method prepares the novel process of human albumin |
| CN1281729A (en) * | 1999-07-22 | 2001-01-31 | 李六金 | Immunoglobulin injection for dog and cat |
| CN1296851A (en) * | 2000-11-18 | 2001-05-30 | 杨盛华 | Antivirus immune globulin for dog, fox and marten and its preparing process |
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2004
- 2004-02-10 CN CNB2004100395712A patent/CN100569287C/en not_active Expired - Lifetime
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1042716A (en) * | 1988-11-17 | 1990-06-06 | 中国医学科学院输血研究所 | Rivanol method prepares the novel process of human albumin |
| CN1281729A (en) * | 1999-07-22 | 2001-01-31 | 李六金 | Immunoglobulin injection for dog and cat |
| CN1296851A (en) * | 2000-11-18 | 2001-05-30 | 杨盛华 | Antivirus immune globulin for dog, fox and marten and its preparing process |
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| 人血狂犬病锡设球蛋白的研制. 宣松林.中国生物制品学杂志，第7卷第1期. 1994 * |
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