CN101953849A - Method for preparing anti-duck viral hepatitis transfer factor - Google Patents

Method for preparing anti-duck viral hepatitis transfer factor Download PDF

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CN101953849A
CN101953849A CN2010105116627A CN201010511662A CN101953849A CN 101953849 A CN101953849 A CN 101953849A CN 2010105116627 A CN2010105116627 A CN 2010105116627A CN 201010511662 A CN201010511662 A CN 201010511662A CN 101953849 A CN101953849 A CN 101953849A
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viral hepatitis
duck viral
transfer factor
spleen
duck
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CN101953849B (en
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吴红云
韩改会
李建正
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Henan Hou Yi bioengineering Limited by Share Ltd
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Zhengzhou Houyi Pharmaceutical Co Ltd
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Abstract

The invention discloses a method for preparing an anti-duck viral hepatitis transfer factor, which comprises the following steps of: firstly, inoculating duck viral hepatitis in a chick embryo allantoic cavity for multiplication and extracting a virus antigen from the chick embryo allantoic cavity; secondly, immunizing a healthy pig body with the extracted virus antigen; and thirdly, extracting the anti-duck viral hepatitis transfer factor from the spleen of the immune pig. In the invention, specific transfer factor is extracted from duck viral hepatitis viruses and can effectively suppress the duck viral hepatitis viruses and protect the cells of a normal body from being infected with the duck viral hepatitis viruses, so that the duck viral hepatitis is prevented and treated radically. Moreover, the preparation method of the invention is simple and feasible, short in production time and low in cost and has a promising application prospect.

Description

A kind of preparation method of anti-duck viral hepatitis transfer factor
Technical field
The present invention relates to a kind of preparation method of anti-duck viral hepatitis transfer factor.
Background technology
Duck viral hepatitis is the viral infectious of bufflehead height lethal, and primary disease finds in the U.S. at first, finds primary disease successively in many foster ducks country such as Britain, Canada, Germany thereafter.The feature of primary disease is that morbidity is anxious, propagates soon, and the mortality rate height faces and examines the performance opisthotonus, and case is changed to hepatitis and hemorrhage.Primary disease is sung to duckery and is caused great economic loss.Though the medicine or the preparation of some anti-duck viral hepatitiss have been arranged at present, can not fundamentally prevent duck viral hepatitis.
Summary of the invention
The preparation method that the purpose of this invention is to provide the anti-duck viral hepatitis transfer factor of a species specificity, to suppress duck viral hepatitis virus, protection normal body cell is avoided the infection of duck viral hepatitis, reaches the fundamentally purpose of prevention.
In order to realize above purpose, the technical solution adopted in the present invention is: a kind of preparation method of anti-duck viral hepatitis transfer factor, with breeding in duck viral hepatitis inoculation and the chick embryo allantoic cavity, therefrom extract virus antigen; Then the virus antigen that extracts is carried out immunity on the health pig body, the spleen extraction from immune swine obtains anti-duck viral hepatitis transfer factor again.
Preparation method concrete steps of the present invention are as follows:
1) the antigenic preparation of DHV: duck viral hepatitis is inoculated in the chick embryo allantoic cavity of 9 ages in days-10 age in days and breeds, 33-37 ℃, cultivate 72-96h, get chick embryo allantoic liquid, in packing in the centrifuge tube of sterilization, centrifugal after the ultrasonic Treatment, discard precipitation and get supernatant, obtain gross virus antigen;
2) extraction of virus antigen: the sucrose solution of in centrifuge tube, carefully laying 20%-66% variable concentrations gradient successively, on liquid level of solution, add crude antigen then, the addition of crude antigen is 20% of each gradient sucrose solution volume, through centrifugal treating, the virus band that careful sucking-off occurs in the saccharose gradient layer, dialysis is removed sucrose and is obtained virus antigen;
3) virus antigen immune swine: select healthy immune swine, carry out subcutaneous multi-point injection immunity with the virus antigen of said extracted, immunity at least 4 times, each immunity is a week at interval, and after last immune 7-10 days, it is stand-by to get the freezing preservation of spleen;
4) from the spleen of immune swine, extract anti-duck viral hepatitis transfer factor:
Figure 2010105116627100002DEST_PATH_IMAGE002
The spleen of immune swine is removed the tissues such as fascia, fat on surface, the reuse normal saline flushing shreds spleen then, adds the normal saline of 2 times of-4 times of volumes, makes the pig spleen homogenate being lower than under 10 ℃ the condition to smash to pieces with high-speed tissue mashing machine;
Figure 2010105116627100002DEST_PATH_IMAGE004
The pig spleen homogenate is carried out ultrasonic Treatment ,-60 ℃ of-80 ℃ of quick freezing, 30 ℃ of-37 ℃ of thawings of water-bath, alternate freezing and thawing 4～6 times all uses the tissue homogenate high-speed homogenization to pulverize behind each the thawing; The freezing normal saline that adds 2 times of volumes then, the sanguine liquid in centrifuging and taking upper strata, reuse sand core funnel filters, and gets filtrate;
Figure 2010105116627100002DEST_PATH_IMAGE006
Use the hydrochloric acid adjust pH to 5.0-6.0 filtrate, supernatant filters with 0.2 μ m hollow fiber column earlier, filter liquor reuse molecular cut off is 6000 daltonian hollow fiber column ultrafilter, collect filter liquor, the filtrate of gained is carried out aseptic filtration with 0.22 μ m filter membrane, promptly obtain anti-duck viral hepatitis transfer factor.
Step 2) described sucrose solution is divided into 4 sucrose concentration gradients that increase progressively, and is respectively 20-25%, 30-45%, 40-55%, 60-66%.
Step 3) is described gets before the immune swine spleen, gets immune swine venous blood, detects the special viral antibody of venous blood with ELISA method or immunofluorescence method and tires, the virus-specific immunizing potency appears and after, get the immune swine spleen again.
Transfer factor is a kind of low molecular weight polypeptide-nucleotide complex that can shift sensitization information that the T lymphocyte discharges, it can be transferred to certain specific cellular immune function of donor the normal lymphocyte of normal receptor specifically, participate in the immunoreation of body, improve cell immune function of human body, the infection of opposing fungus, virus, antibacterial etc.; Promote the transfer factor secretory action of bone-marrow-derived lymphocyte simultaneously, can discharge interferon and interleukin by inducing cell, thereby strengthen the immunologic function of receptor, be described as the triggering agent of T cytoactive.The transfer factor molecular weight is little, nontoxic, no antigen, do not play anaphylaxis, and transfer factor and can to surmount kind be the boundary advantages of application does not create antagonism.
The present invention adopts duck viral hepatitis virus to extract specific transfer factor; this transfer factor can effectively suppress duck viral hepatitis virus; can protect the normal body cell to avoid the infection of duck viral hepatitis virus, fundamentally prevent the generation of duck viral hepatitis.In addition, preparation method of the present invention is simple, and the production time is short, and cost is low, has good application prospects.
The specific embodiment
Embodiment 1
It is example that present embodiment adopts duck 1 type viral hepatitis, prepares anti-duck viral hepatitis transfer factor, and the step of this method is as follows:
1) the antigenic preparation of DHV: duck 1 type viral hepatitis is inoculated in the chick embryo allantoic cavity of 9 ages in days and breeds, 33 ℃, cultivate 72h, get chick embryo allantoic liquid, in packing in the centrifuge tube of sterilization, use ultrasonic echography 10min(50Hz), the centrifugal 20min of 5000rpm/min, discard precipitation and get supernatant, obtain gross virus antigen;
2) extraction of virus antigen: in centrifuge tube, carefully lay 20%, 30%, 40% successively, the sucrose solution of 66% 4 gradients, each gradient sucrose solution is 1ml, on liquid level of solution, add crude antigen 0.2ml at last, through the centrifugal 2.5h of 40000r/min, at the second layer one virus band is arranged, careful sucking-off, dialysis goes sucrose to obtain virus antigen;
3) duck 1 type viral hepatitis antigen immune pig: select healthy immune swine, virus antigen with said extracted carries out subcutaneous multi-point injection immunity, immunity is 4 times altogether, each immunity is a week at interval, last immunity was got porcine vein after 7 days, detect the specific antibody titres of above-mentioned virus with ELISA method or immunofluorescence method, occur slaughtering animal after the virus-specific immunizing potency, get spleen and place-20 ℃ of preservations stand-by;
4) from immune swine, extract anti-duck pestilence transfer factor:
The spleen of immune swine is removed tissues such as surperficial fascia, fat, the reuse normal saline flushing, then spleen is shredded, the normal saline that adds 2 times of volumes, adopt normal saline can prevent the active destroyed of transfer factor, smash to pieces with high-speed tissue mashing machine under 4 ℃ condition, 1500rpm smashs to pieces 3 times, each 3min makes the pig spleen homogenate;
Figure 706932DEST_PATH_IMAGE004
The pig spleen homogenate is carried out ultrasound wave (4KW, 3KHz) handle 5min ,-60 ℃ of quick freezing, 30 ℃ of thawings of water-bath, alternate freezing and thawing 4 times all uses the tissue homogenate high-speed homogenization to pulverize behind each the thawing; The freezing normal saline that adds 2 times of volumes then, 4 ℃ of centrifugal 20min of 5000r/min get the sanguine liquid in upper strata, and the sand core funnel of reuse G2 filters, and gets filtrate;
Figure 424352DEST_PATH_IMAGE006
Filtrate with 0.1% hydrochloric acid adjust pH to 5.0, can effectively be removed foreign protein, make the supernatant after centrifugal purer; Supernatant filters with 0.2 μ m hollow fiber column earlier, can effectively ensure the pure property and the activity of transfer factor, has shortened preparation time simultaneously; Filter liquor reuse molecular cut off is 6000 daltonian hollow fiber column ultrafilter, collects filter liquor, and the filtrate of gained is carried out aseptic filtration with 0.22 μ m filter membrane, promptly obtains anti-duck 1 type viral hepatitis transfer factor extracting solution.
Embodiment 2
It is example that present embodiment adopts duck 1 type viral hepatitis, prepares anti-duck viral hepatitis transfer factor, and the step of this method is as follows:
1) the antigenic preparation of DHV: duck 1 type viral hepatitis is inoculated in the chick embryo allantoic cavity of 9 ages in days and breeds, 35 ℃, cultivate 84h, get chick embryo allantoic liquid, in packing in the centrifuge tube of sterilization, use ultrasonic echography 10min(50Hz), the centrifugal 20min of 5000rpm/min, discard precipitation and get supernatant, obtain gross virus antigen;
2) extraction of virus antigen: in centrifuge tube, carefully lay 20%, 30%, 45% successively, the sucrose solution of 60% 4 gradients, each gradient sucrose solution is 1ml, on liquid level of solution, add crude antigen 0.2ml at last, through the centrifugal 2.5h of 40000r/min, at the second layer one virus band is arranged, careful sucking-off, dialysis goes sucrose to obtain virus antigen;
3) duck 1 type viral hepatitis antigen immune pig: select healthy immune swine, virus antigen with said extracted carries out subcutaneous multi-point injection immunity, immunity is 4 times altogether, each immunity is a week at interval, last immunity was got porcine vein after 8 days, detect the specific antibody titres of above-mentioned virus with ELISA method or immunofluorescence method, occur slaughtering animal after the virus-specific immunizing potency, get spleen and place-20 ℃ of preservations stand-by;
4) from immune swine, extract anti-duck pestilence transfer factor:
Figure 819561DEST_PATH_IMAGE002
The spleen of immune swine is removed tissues such as surperficial fascia, fat, the reuse normal saline flushing, then spleen is shredded, the normal saline that adds 3 times of volumes, adopt normal saline can prevent the active destroyed of transfer factor, smash to pieces with high-speed tissue mashing machine under 6 ℃ condition, 2500rpm smashs to pieces 4 times, each 4min makes the pig spleen homogenate;
Figure 562828DEST_PATH_IMAGE004
The pig spleen homogenate is carried out ultrasound wave (4KW, 3KHz) handle 7min ,-70 ℃ of quick freezing, 35 ℃ of thawings of water-bath, alternate freezing and thawing 5 times all uses the tissue homogenate high-speed homogenization to pulverize behind each the thawing; The freezing normal saline that adds 2 times of volumes then, 4 ℃ of centrifugal 20min of 5000r/min get the sanguine liquid in upper strata, and the sand core funnel of reuse G2 filters, and gets filtrate;
Figure 322973DEST_PATH_IMAGE006
Filtrate with 0.1% hydrochloric acid adjust pH to 5.5, can effectively be removed foreign protein, make the supernatant after centrifugal purer; Supernatant filters with 0.2 μ m hollow fiber column earlier, can effectively ensure the pure property and the activity of transfer factor, has shortened preparation time simultaneously; Filter liquor reuse molecular cut off is 6000 daltonian hollow fiber column ultrafilter, collects filter liquor, and the filtrate of gained is carried out aseptic filtration with 0.22 μ m filter membrane, promptly obtains anti-duck 1 type viral hepatitis transfer factor extracting solution.
Embodiment 3
It is example that present embodiment adopts duck 1 type viral hepatitis, prepares anti-duck viral hepatitis transfer factor, and the step of this method is as follows:
1) the antigenic preparation of DHV: duck 1 type viral hepatitis is inoculated in the chick embryo allantoic cavity of 10 ages in days and breeds, 37 ℃, cultivate 96h, get chick embryo allantoic liquid, in packing in the centrifuge tube of sterilization, use ultrasonic echography 10min(50Hz), the centrifugal 20min of 5000rpm/min, discard precipitation and get supernatant, obtain gross virus antigen;
2) extraction of virus antigen: in centrifuge tube, carefully lay 25%, 45%, 55% successively, the sucrose solution of 65% 4 gradients, each gradient sucrose solution is 1ml, on liquid level of solution, add crude antigen 0.2ml at last, through the centrifugal 2.5h of 40000r/min, at the second layer one virus band is arranged, careful sucking-off, dialysis goes sucrose to obtain virus antigen;
3) duck 1 type viral hepatitis antigen immune pig: select healthy immune swine, virus antigen with said extracted carries out subcutaneous multi-point injection immunity, immunity is 4 times altogether, each immunity is a week at interval, last immunity was got porcine vein after 10 days, detect the specific antibody titres of above-mentioned virus with ELISA method or immunofluorescence method, occur slaughtering animal after the virus-specific immunizing potency, get spleen and place-20 ℃ of preservations stand-by;
4) from immune swine, extract anti-duck pestilence transfer factor:
Figure 527690DEST_PATH_IMAGE002
The spleen of immune swine is removed tissues such as surperficial fascia, fat, the reuse normal saline flushing, then spleen is shredded, the normal saline that adds 4 times of volumes, adopt normal saline can prevent the active destroyed of transfer factor, smash to pieces with high-speed tissue mashing machine under 2 ℃ condition, 3000rpm smashs to pieces 5 times, each 5min makes the pig spleen homogenate;
Figure 726590DEST_PATH_IMAGE004
The pig spleen homogenate is carried out ultrasound wave (4KW, 3KHz) handle 10min ,-80 ℃ of quick freezing, 37 ℃ of thawings of water-bath, alternate freezing and thawing 6 times all uses the tissue homogenate high-speed homogenization to pulverize behind each the thawing; The freezing normal saline that adds 2 times of volumes then, 4 ℃ of centrifugal 20min of 5000r/min get the sanguine liquid in upper strata, and the sand core funnel of reuse G2 filters, and gets filtrate;
Filtrate with 0.1% hydrochloric acid adjust pH to 6.0, can effectively be removed foreign protein, make the supernatant after centrifugal purer; Supernatant filters with 0.2 μ m hollow fiber column earlier, can effectively ensure the pure property and the activity of transfer factor, has shortened preparation time simultaneously; Filter liquor reuse molecular cut off is 6000 daltonian hollow fiber column ultrafilter, collects filter liquor, and the filtrate of gained is carried out aseptic filtration with 0.22 μ m filter membrane, promptly obtains anti-duck 1 type viral hepatitis transfer factor extracting solution.
Experimental example 1: animal protection test
Implement the duckling that 2000 7 ages in days are got in the experiment of 1 animal protection, be divided into experimental group and matched group, every group each 1000.Counteracting toxic substances dosage is 1.0 * 10 7CFU/ only, the 3rd day experimental group injected anti-duck viral hepatitis transfer factor behind the counteracting toxic substances, every intramuscular injection 0.1ml every day 1 time, matched group injection equivalent distilled water, observe duckling symptom, observing 5 days continuously, is the therapeutic effect that index reflects anti-duck viral hepatitis transfer factor with mental status and mortality rate.The result shows: duckling 100% death of matched group injection equivalent distilled water, inject anti-duck viral hepatitis transfer factor mortality rate 8.1%.
Implement the experiment of 2 animal protections: get the duckling of 200 7 ages in days, be divided into experimental group and matched group, every group each 100.Counteracting toxic substances dosage is 1.0 * 10 7CFU/ only, the 3rd day experimental group injected anti-duck viral hepatitis transfer factor behind the counteracting toxic substances, every intramuscular injection 0.1ml every day 1 time, matched group injection equivalent distilled water, observe duckling symptom, observing 5 days continuously, is the therapeutic effect that index reflects anti-duck viral hepatitis transfer factor with mental status and mortality rate.The result shows: duckling 100% death of matched group injection equivalent distilled water, inject anti-duck viral hepatitis transfer factor death in rate 8.0%.
Implement the experiment of 2 animal protections: get the duckling of 200 7 ages in days, be divided into experimental group and matched group, every group each 100.Counteracting toxic substances dosage is 1.0 * 10 7CFU/ only, the 3rd day experimental group injected anti-duck viral hepatitis transfer factor behind the counteracting toxic substances, every intramuscular injection 0.1ml every day 1 time, matched group injection equivalent distilled water, observe duckling symptom, observing 5 days continuously, is the therapeutic effect that index reflects anti-duck viral hepatitis transfer factor with mental status and mortality rate.The result shows: duckling 100% death of matched group injection equivalent distilled water, inject anti-duck viral hepatitis transfer factor death 8.0%.
Experimental example 2
This experimental example adopts various conventional methods that anti-duck viral hepatitis transfer factor of the present invention is detected:
1, ultraviolet spectrophotometry: this product has a high absworption peak at the 250.0-252.0nm place, and ABS260/ABS280＞2.0.
2, mark liquid is faint yellow, and pH value is between 6.0-6.5.
3, bacteriological detection: this kind need not oxygen, anaerobism, saprophytic bacteria and fungus and exists.
4,20% sulfosalicylic acid detects: this product does not have muddy chicken deposited phenomenon, illustrates that albumino reaction is all negative, and it does not contain the large protein molecule.
5, the mensuration of content of peptides: it is 1.530mg/ml that this product is measured content of peptides through biuret method.
6, Determination of Nucleic Acid Content: it is 596.89 μ g/ml that this product is measured nucleic acid content through orcin method.
7, E-rosette formation rate relatively: there is the Mus erythrocyte receptor on duck peripheral blood lymphocyte surface, and can be with it in conjunction with forming the E-rosette.And transfer factor is combined with certain facilitation to lymphocyte and rat are erythrocytic.The E-rosette formation rate average out to 30.1% of this product is than increase 24.6%(P≤0.01 of matched group).
8, safety verification: get 200 of healthy mices, be divided into 2 groups of normal group and immune group, normal group allows its oral distilled water, immune group allows its oral this product concentrated solution (be equivalent to normal oral dose 50 times), observes the variation of mice, result: the survival of immune group 100%, and there are not any poison, anaphylaxis, death condition illustrates that this product is safe, does not have any toxicity and side effect.
9, do the skin test experiment with duck viral hepatitis antigen and detect, faint positive reaction is arranged, illustrate that this product has good transfer activity and specificity.
By above experimental example, we can see that the sick transfer factor of the anti-duck viral hepatitis that is extracted can suppress duck viral hepatitis virus safely and effectively, can protect normal body cell to avoid the infection of duck viral hepatitis.Use transfer factor of the present invention can prevent the generation of duck viral hepatitis.

Claims (4)

1. the preparation method of an anti-duck viral hepatitis transfer factor is characterized in that: with breeding in duck viral hepatitis inoculation and the chick embryo allantoic cavity, therefrom extract virus antigen; Then the virus antigen that extracts is carried out immunity on the health pig body, the spleen extraction from immune swine obtains anti-duck viral hepatitis transfer factor again.
2. the preparation method of anti-duck viral hepatitis transfer factor according to claim 1 is characterized in that: its concrete steps are as follows:
1) the antigenic preparation of DHV: duck viral hepatitis is inoculated in the chick embryo allantoic cavity of 9 ages in days-10 age in days and breeds, 33-37 ℃, cultivate 72-96h, get chick embryo allantoic liquid, in packing in the centrifuge tube of sterilization, centrifugal after the ultrasonic Treatment, discard precipitation and get supernatant, obtain gross virus antigen;
2) extraction of virus antigen: the sucrose solution of in centrifuge tube, carefully laying 20%-66% variable concentrations gradient successively, on liquid level of solution, add crude antigen then, the addition of crude antigen is 20% of each gradient sucrose solution volume, through centrifugal treating, the virus band that careful sucking-off occurs in the saccharose gradient layer, dialysis is removed sucrose and is obtained virus antigen;
3) virus antigen immune swine: select healthy immune swine, carry out subcutaneous multi-point injection immunity with the virus antigen of said extracted, immunity at least 4 times, each immunity is a week at interval, and after last immune 7-10 days, it is stand-by to get the freezing preservation of spleen;
4) from the spleen of immune swine, extract anti-duck viral hepatitis transfer factor:
Figure 2010105116627100001DEST_PATH_IMAGE002
The spleen of immune swine is removed the tissues such as fascia, fat on surface, the reuse normal saline flushing shreds spleen then, adds the normal saline of 2 times of-4 times of volumes, makes the pig spleen homogenate being lower than under 10 ℃ the condition to smash to pieces with high-speed tissue mashing machine;
Figure 2010105116627100001DEST_PATH_IMAGE004
The pig spleen homogenate is carried out ultrasonic Treatment ,-60 ℃ of-80 ℃ of quick freezing, 30 ℃ of-37 ℃ of thawings of water-bath, alternate freezing and thawing 4～6 times all uses the tissue homogenate high-speed homogenization to pulverize behind each the thawing; The freezing normal saline that adds 2 times of volumes then, the sanguine liquid in centrifuging and taking upper strata, reuse sand core funnel filters, and gets filtrate;
Use the hydrochloric acid adjust pH to 5.0-6.0 filtrate, supernatant filters with 0.2 μ m hollow fiber column earlier, filter liquor reuse molecular cut off is 6000 daltonian hollow fiber column ultrafilter, collect filter liquor, the filtrate of gained is carried out aseptic filtration with 0.22 μ m filter membrane, promptly obtain anti-duck viral hepatitis transfer factor.
3. the preparation method of anti-duck viral hepatitis transfer factor according to claim 2 is characterized in that: step 2) described sucrose solution is divided into 4 sucrose concentration gradients that increase progressively, and is respectively 20-25%, 30-45%, 40-55%, 60-66%.
4. the preparation method of anti-duck viral hepatitis transfer factor according to claim 2, it is characterized in that: step 3) is described gets before the immune swine spleen, get immune swine venous blood, detect the special viral antibody of venous blood tires with ELISA method or immunofluorescence method, after the virus-specific immunizing potency occurring, get the immune swine spleen again.
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CN102697808A (en) * 2012-05-04 2012-10-03 郑州后羿制药有限公司 In-vitro preparation method of transfer factors capable of resisting duck viral hepatitis
CN102697811A (en) * 2012-05-04 2012-10-03 郑州后羿制药有限公司 Method for preparing transfer factors capable of resisting gosling plague virus
CN103599534A (en) * 2013-10-24 2014-02-26 福州派生特生物科技有限公司 Preparation method and use of poultry vaccine-specific pig spleen transfer factor
CN104162161A (en) * 2013-07-24 2014-11-26 河南联合英伟饲料有限公司 Composition for resisting duck viral hepatitis, freeze-dried powder, and preparation method and use of freeze-dried powder
CN104162160A (en) * 2013-07-24 2014-11-26 河南联合英伟饲料有限公司 Antiviral composition for feed additive, antiviral freeze-dried powder for feed additive and preparation method and use of antiviral freeze-dried powder for feed additive
CN104162159A (en) * 2013-07-24 2014-11-26 河南联合英伟饲料有限公司 Composition for resisting canine parvovirus, freeze-dried powder, and preparation method and use of freeze-dried powder
CN105497867A (en) * 2014-09-26 2016-04-20 天津嘉瑞生物科技有限公司 Preparation method for transfer factor capable of resisting avian infectious laryngotracheitis virus
CN105560282A (en) * 2014-10-14 2016-05-11 天津嘉瑞生物科技有限公司 Preparation method of anti-duck hepatitis virus transfer factor
CN105561287A (en) * 2014-10-14 2016-05-11 天津嘉瑞生物科技有限公司 Preparation method of anti-duck hepatitis virus transfer factor
CN108865885A (en) * 2018-07-05 2018-11-23 广州铭康生物工程有限公司 One kind is for the efficient device for trapping of cell culture and its retention method

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CN102697811A (en) * 2012-05-04 2012-10-03 郑州后羿制药有限公司 Method for preparing transfer factors capable of resisting gosling plague virus
CN102697811B (en) * 2012-05-04 2014-02-05 郑州后羿制药有限公司 Method for preparing transfer factors capable of resisting gosling plague virus
CN102697808B (en) * 2012-05-04 2014-02-05 郑州后羿制药有限公司 In-vitro preparation method of transfer factors capable of resisting duck viral hepatitis
CN102697808A (en) * 2012-05-04 2012-10-03 郑州后羿制药有限公司 In-vitro preparation method of transfer factors capable of resisting duck viral hepatitis
CN104162161B (en) * 2013-07-24 2016-09-21 河南联合英伟饲料有限公司 A kind of anti-duck viral hepatitis compositions, lyophilized powder, preparation method and application
CN104162161A (en) * 2013-07-24 2014-11-26 河南联合英伟饲料有限公司 Composition for resisting duck viral hepatitis, freeze-dried powder, and preparation method and use of freeze-dried powder
CN104162160A (en) * 2013-07-24 2014-11-26 河南联合英伟饲料有限公司 Antiviral composition for feed additive, antiviral freeze-dried powder for feed additive and preparation method and use of antiviral freeze-dried powder for feed additive
CN104162159A (en) * 2013-07-24 2014-11-26 河南联合英伟饲料有限公司 Composition for resisting canine parvovirus, freeze-dried powder, and preparation method and use of freeze-dried powder
CN104162160B (en) * 2013-07-24 2016-09-21 河南联合英伟饲料有限公司 Feed additive antiviral composition, lyophilized powder, preparation method and application
CN104162159B (en) * 2013-07-24 2016-09-21 河南联合英伟饲料有限公司 A kind of anti-dog parvovirus compositions, lyophilized powder, preparation method and application
CN103599534A (en) * 2013-10-24 2014-02-26 福州派生特生物科技有限公司 Preparation method and use of poultry vaccine-specific pig spleen transfer factor
CN103599534B (en) * 2013-10-24 2015-07-08 福州派生特生物科技有限公司 Preparation method and use of poultry vaccine-specific pig spleen transfer factor
CN105497867A (en) * 2014-09-26 2016-04-20 天津嘉瑞生物科技有限公司 Preparation method for transfer factor capable of resisting avian infectious laryngotracheitis virus
CN105561287A (en) * 2014-10-14 2016-05-11 天津嘉瑞生物科技有限公司 Preparation method of anti-duck hepatitis virus transfer factor
CN105560282A (en) * 2014-10-14 2016-05-11 天津嘉瑞生物科技有限公司 Preparation method of anti-duck hepatitis virus transfer factor
CN108865885A (en) * 2018-07-05 2018-11-23 广州铭康生物工程有限公司 One kind is for the efficient device for trapping of cell culture and its retention method
CN108865885B (en) * 2018-07-05 2021-03-16 广州铭康生物工程有限公司 Cell culture interception device and cell culture interception method

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