CN1382736A - Yolk antibody and antigen of pig's infective enterogastritis virus and its preparing process - Google Patents
Yolk antibody and antigen of pig's infective enterogastritis virus and its preparing process Download PDFInfo
- Publication number
- CN1382736A CN1382736A CN 02103667 CN02103667A CN1382736A CN 1382736 A CN1382736 A CN 1382736A CN 02103667 CN02103667 CN 02103667 CN 02103667 A CN02103667 A CN 02103667A CN 1382736 A CN1382736 A CN 1382736A
- Authority
- CN
- China
- Prior art keywords
- yolk
- antibody
- yolk antibody
- transmissible gastro
- virus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
A yolk antibody for pig's infective enterogastritis virus is prepared through reproducing the said virus on pig's testis cell, applying it as immunogen to the layer, collecting yolk and purifying. When it is applied to pig, the comparison between experimental and reference groups shows that it has high effect on preventing and cuting pig's infective enterogastritis.
Description
Technical field
The present invention relates to the antibody of a kind of transmissible gastro-enteritis virus and antigen and preparation method thereof.
Background technology
(porcine transmissible gastroenteritis virus TGEV) is the member of coronaviridae, coronavirus genus to transmissible gastro-enteritis virus.TGEV virus can infect the pig of institute's has age, especially cause that following piglet vomitted, seriously suffered from diarrhoea and high mortality (common 100%) two ages in week, it is the boar height contact severe intestinal transmissible disease that worldwide extensively exists at present, usually take place repeatedly to bring very big harm to pig industry with popular on the pig farm.(PensaertM?B.Disease?of?Swine.Seventh?edition.Iowa?State?University?Press.USA.1992，305-338)。
At present the control of this disease still there are not especially effectively means.Feasible methods of treatment only is a symptomatic treatment, reduces hunger, dehydration and oxypathy.Immunoprophylaxis report about TGEV is more, the active immunity of existing use strong poison of TGEV or weak poison, also having feeds contains passive immunization (the Porter P. of TGEV antibody milk, Allen W.D.Classes of immunoglobulinsrelated to immunity in the pig:A review.J.AM.Vet.Med.Assoc.1972,160:511-518), also have novel subunit vaccine, live recombinant vectors vaccine etc.In view of TGEV in enteron aisle internal adsorption and propagation, whether transmissible gastroenteritis of swine takes place with the height of enteron aisle local immunity level closely related, thereby the local immunity of enteron aisle is particularly important to control TGEV, the height of secretory IgA level often reflects the effect of immunity, the serum antibody level may be very high behind the TGEV vaccine immunity pig body that uses at present, and enteron aisle local immunity level is relatively poor, can not reach the purpose that stops TGEV to infect.Though and the milk immunity can be protected the infection of newborn piglet opposing TGEV, promptly lose protection after the ablactation.
Yolk antibody can be resisted the hydrochloric acid in gastric juice barrier of young animal, the digestion of trypsinase and curdled milk proteolytic enzyme in the opposing enteron aisle.This kind antibody titer length of holding time, chemical property is stable, can heat-resisting, sour, alkali, but 4 ℃ of long storage time.Yolk antibody also has preparation process than serum antibody is simple, time saving and energy saving, output is high, cost is low, be easy to many advantages such as acquisition and purification.Usually produce high antibody of tiring behind the former immune chicken body of virus immunity in yolk, and immunizing rabbit etc. are used to prepare the common antibody titer height of serum antibody, results serum antibody amount is less, and cost is higher.(Ntakarutimana?V.，DemedtsP.，Sande?M.V.et?al：A?simple?and?economical?strategy?for?downstream?processing?of?specificantibodies?to?human?transferring?from?egg?yolk.J.Immun.Meth.1992，153：133-140。BizhanovG.，Vyshniauskis?G：A?comparison?of?three?methods?for?extracting?IgY?from?the?egg?yolk?of?hensimmunized?with?Sendai?virus.Vet.Res.Comm.2000，24：103-113。Luo Han high post: the application of high immunity yolk antibody and prospect thereof. Jiangsu agricultural sciences .1997,5:59-63).But the transmissible gastro-enteritis virus yolk antibody does not appear in the newspapers as yet.
Summary of the invention
The technical problem to be solved in the present invention is a kind of anti-TGEV yolk antibody of development, can directly act on the enteron aisle of animal after this antibody oral administration, stops the purpose of TGEV in enteron aisle absorption and propagation thereby reach.
Technical scheme of the present invention is that transmissible gastro-enteritis virus is bred on the pig testis cell after the rejuvenation in the pig body, be used to prepare immunogen, the non-immune laying hen of immunity is collected yolk and carries out yolk antibody detections of tiring, the yolk purification yolk antibody of selecting height to tire.Directly act on intestine of young pigs after this antibody oral administration, infect with treatment and prevention TGEV.
Concrete grammar is:
One .TGEV antigen prepd:
1. piglet feeds and attacks poison
Do not eat the oral TGEV virus of suckling piglet liquid just for 1 age in days, catch and kill piglet after 48 hours, take out whole small intestines, be used for extracting and purifying TGEV.
2.TGEV extraction and purification
Small intestine is shredded the back fully grind, add 0.01MPBS (pH7.5) dilution, multigelation, centrifugal, get supernatant liquor and add 6%PEG-6000 and 2%NaCl, centrifugal again after the placement, precipitation is resuspended among the PBS, getting supernatant liquor after centrifugal carries out ultracentrifugation.Precipitation is resuspended among the PBS, is the purified virus particle.
3.TGEV on cell, breed
Cultivate pig testis (ST) cell with MEM nutrient solution (containing 5% calf serum), when cell monolayer grows to 75% when above, getting the ultracentrifugation purified virus is inoculated on the cell monolayer, and the adding pancreatin is beneficial to the absorption of virus, if 72 hours the postoperative infection cytopathy (CPE) not obvious, then the centrifuging and taking supernatant liquor continues blind passage on cell behind the harvested cell suspension multigelation, blind passage to the third generation begins to occur CPE, this moment is cells infecteds in a large number, the harvested cell suspension, centrifugal behind the multigelation, supernatant liquor is used as antigen after concentrating.
Two. the yolk antibody preparation
1. animal immune
(1) immunity is prepared: above-mentioned concentrated antigen liquid eggs in vain with the PBS dilution, and is exempted to use immunogen as head after the emulsification of equivalent Freund's complete adjuvant; With after the emulsification of equivalent Freund's incomplete adjuvant as two exempting from, three exempting from and four exempt to use immunogen.
(2) immunity: the immunogen with above-mentioned preparation is carried out immunity to non-immunization laying hen, and other establishes not immune blank group, raises according to a conventional method in common hen house.
Immune programme for children is:
Head exempts from preceding 5 days to every thoracic muscle multi-point injection 0.5ml " Vaccinum Pertussis et Toxoidum Diphthero Tetanicum Adsorbatum, diphtheria, Toxoid,tetanus " mixed preparation, to stimulate organism immune response.Every thoracic muscle multi-point injection immunogen 1ml when head exempts from carries out two exempting from (the immunity amount is identical when just exempting from) after two weeks, exempt from (the immunity amount doubles) every carrying out 3 two weeks again, carries out the 4th immunity (the immunity amount is redoublingd) again after one month.
2. yolk liquid preparation
Two exempt from the back began to collect the egg that produces in one week, isolated yolk, mixed back homogenate with equivalent PBS, was prepared into yolk liquid and decided antibody titer to descend pacing.
3. antibody titer detects
The TGEV yolk antibody that obtains proves by the antigen antibody reaction in the agar diffusion test.(concrete grammar is with reference to the veterinary immunology experiment instruction, and Agricultural University Of Nanjing edits, Shanghai science tech publishing house, 1984).Detect antigen in the agar diffusion and be super virus from purification, because the immunogen of laying hen immunity usefulness is from the cell infection thing, wherein impure composition may be cell debris, and there is not cellular constituent in the detection antigen, if this detection antigen and yolk antibody react, containing at detecting antigen in the provable yolk antibody is the antibody of TGEV.
Preparation agar gel: in hole around the adding agar gel behind the yolk antibody liquid usefulness PBS doubling dilution, add antigen in the interstitial hole through the ultracentrifugation purifying, put in the wet box in 37 ℃ of incubators reactions 24 hours, with high dilution yolk liquid that positive precipitation band occurs antibody titer for this yolk liquid.
Detected result: two exempt to gather in one week of back yolk detects that to tire be 1: 8, regathers month antibody titer of yolk and peaks 1: 64.The antibody titer experimental data has further proved the active height of yolk antibody.
4. yolk antibody is purified
Yolk antibody is purified and is adopted polyoxyethylene glycol (PEG-6000) precipitator method.(method is referring to Polson A., etal.Improvement in the isolation of IgY from the yolks of eggs laid by immunized hens.Immun.Invest., and 1985,14 (4), 323-327).
5. with the yolk antibody liquid freeze-drying of purifying, be stored in 4 ℃.
Three .TGEV yolk antibody therapeutic tests
Laboratory animal is that 1 age in days is not eaten suckling piglet just.The test group animal is used TGEV virus oral challenge, the oral then yolk antibody of the present invention of feeding.Other establish attack poison not with the antibody group, do not attack poison not with the antibody group, do not attack poison in contrast with 3 groups of antibody groups.Judge the result of treatment of TGEV yolk antibody according to each group clinical symptom and mortality ratio.
Test-results shows: the grice diarrhoea that TGEV yolk antibody provided by the invention causes TGEV has the obvious treatment effect, any toxic side effect do not occur.
Appended drawings is yolk antibody of the present invention and chicken serum antibody titer change curve, and wherein, Fig. 1 is No. 1 chicken serum antibody and yolk antibody change curve; Fig. 2 is No. 2 chicken serum antibody and yolk antibody change curve; Fig. 3 is No. 3 chicken serum antibody and yolk antibody change curve; Fig. 4 is No. 4 chicken serum antibody and yolk antibody change curve; Fig. 5 is No. 5 chicken serum antibody and yolk antibody change curve.
Embodiment
Embodiment 1 transmissible gastro-enteritis virus (TGEV) propagation and antigen prepd
This test selects for use pig testis cell (ST) as cells infected, and adds an amount of trypsinase to promote the adaptation and the infection of virus when virus infection, and test-results shows that the virus infection titre obviously improves, and obvious cytopathy occurs.1. material 1.1TGEV strain isolated is from China Veterinary Drugs Supervisory Inst..1.2 newborn piglet 1 age in days is not eaten first suckling piglet from the Beijing suburb pig farm that does not have TGEV morbidity history.1.3 pig testis cell (ST) is available from China Veterinary Drugs Supervisory Inst..1.4 cell culture medium MEM basic medium is available from GIBCO BRL company, new-born calve serum is available from Hyclone company.2. method 2.1TGEV does not eat after just suckling piglet is bought back at propagation 1 age in days on the pig body, use milk feeding, every two hours feed once, observe after one day, it is oral to get TGEV virus liquid, attack poison and began to occur symptom of diarrhea in back 12 hours, the ight soil deliquescing is thinning, occurs serious yellow watery diarrhea then, apocleisis and skeletonize appear rapidly in piglet, cutd open to 48 hours and to kill the disease pig, take out whole small intestines, be used for extracting and purifying TGEV.2.2TGEV extraction and purification shreds small intestine with scissors, pours in the tissue grinder and fully grinds, and adds 10 times of volume 0.01MPBS (pH7.4) dilution, multigelation three times, 5000rpm
*30min is centrifugal, gets supernatant liquor and adds 6%PEG-6000 and 2%NaCl, and 5000rpm is spent the night in 4 ℃ of placements
*30min is centrifugal, precipitation is resuspended among the PBS of 1/5 stoste volume, and 4 ℃ of placements are spent the night, and resuspended liquid is through 12000
*Get supernatant liquor after 30min is centrifugal and carry out ultracentrifugation, 25000rpm
*2hr (Beckman L7 type ultracentrifuge), precipitation is resuspended among the PBS of 1/10 stoste volume, is the purified virus particle.2.3 breeding with MEM nutrient solution (containing 5% calf serum), TGEV cultivates pig testis (ST) cell on cell, when cell monolayer grows to 75% when above, getting the ultracentrifugation purified virus is inoculated on the cell monolayer after 10 times of PBS dilutions, and adding 50ug/ml pancreatin is beneficial to the absorption of virus, if 72 hours the postoperative infection cytopathy (CPE) not obvious, then the centrifuging and taking supernatant liquor continues blind passage on cell behind the harvested cell suspension multigelation three times, blind passage to the third generation begins to occur CPE, show as cytogamy and round contracting, plaqueization appears in the part cell detachment.Gather in the crops sick cell this moment, inoculates the ST cell in a large number, centrifugal behind the harvested cell suspension, multigelation three times, and supernatant liquor is used as antigen after concentrating.The former TGEV cell toxicant of preparation 1. materials, the 1.1 virus immunities concentrated antigen of embodiment 2 TGEV yolk antibodies is the above, and the mensuration protein concentration is 3.3mg/ml, and it is standby to be diluted to 2.0mg/ml with PBS.1.2 adjuvant Freund's complete adjuvant and Freund's incomplete adjuvant are available from GIBCO BRL company.1.3 7 of the non-immune laying hens of non-immune laying hen 120 ages in days, wherein 5 are used for the TGEV immunity, other 2 as blank, ordinary method is raised.1.4 it is analytical reagent that yolk antibody is purified with reagent PEG-6000, available from positive Xinghua, Suzhou worker institute.2. method 2.1 immunogen preparing are got the former and Freund's complete adjuvant of virus immunity that two syringes are drawn equivalent respectively, the centre connects a thin rubber tubing, by two syringes abundant mixing immunogen of mode to pushing away respectively, be even white emulsion, placement is spent the night not stratified for qualified, exempts to use immunogen as head.Virus immunity is former to prepare immunogen equally with Freund's incomplete adjuvant, as two exempting from, three exempt from, and four exempting to use immunogen.2.2 immune head exempts from preceding 5 days to every thoracic muscle multi-point injection 0.5ml " Vaccinum Pertussis et Toxoidum Diphthero Tetanicum Adsorbatum, diphtheria, Toxoid,tetanus " mixed preparation, to stimulate organism immune response.Every thoracic muscle multi-point injection immunogen 1ml when head exempts from carries out two exempting from (the immunity amount is identical when just exempting from) after two weeks, exempt from (the immunity amount doubles) every carrying out 3 two weeks again, carries out the 4th immunity (the immunity amount is redoublingd) again after one month.2.3 exempting from the back, yolk liquid preparation two one week began to collect egg that immune chicken produces, isolating yolk, is a collector unit by every chicken egg that every two weeks produces, and the yolk of collection mixes back homogenate with equivalent PBS, and the adding minimum of chloroform is anticorrosion, is prepared into yolk liquid in order to measuring antibody titer.Begin blood sampling one week 2.4 the back is exempted from serum antibody extraction two, every immune chicken wings venous blood collection 1ml puts 37 ℃ of incubators and placed 2 hours, put 4 ℃ of refrigerator overnight then, sucking-off serum is used to measure antibody titer, and every the blood sampling of two weeks once, separation of serum is measured serum antibody titer later on.Add 1.5 gram agar powders 2.5 antibody titer detects the preparation 100ml 8% NaCl solution of (agar diffusion method) 2.5.1 agar plate, heating and melting adds 0.2ml phenol mixing, pour glass dish into, make the thick agar plate of 2-3mm, after room temperature was solidified, 4 ℃ of inversions were preserved stand-by.2.5.2 the antibody doubling dilution is made doubling dilution with PBS respectively with serum and yolk, extent of dilution is respectively 2 *, 4 *, 8 *, 16 *, 32 *, 64 * and highly diluted multiple more.2.5.3 the agar diffusion reaction breaks into seven apertures in the human head blossom type hole with punch tool with agar plate, spirit lamp heating back cover, add different dilution antibody on every side in the hole, add the excess of imports in the interstitial hole from the TGEV antigen of purifying, be inverted plate, 37 ℃ of airtight wet box internal reactions 24 to 72 hours produce until precipitation line.With the antibody maximum dilution multiple of observing precipitation line is the antibody titer of test sample.Serum antibody titer and the yolk antibody detected result of tiring sees Table 1.Every chicken serum antibody and the yolk antibody change curve of tiring is seen Fig. 1-Fig. 5.
The immune chicken of table 1 is every two all serum antibodies and yolk antibody titration result
Immune chicken serum antibody and the yolk antibody comparative result of tiring shows:
| Detection time (week) | ????2 | ????4 | ????6 | ????8 | ????10 | ????12 | ????14 | ????16 | |
| ??1 | Serum antibody | ????1∶16 | ??1∶32 | ??1∶64 | ??1∶32 | ??1∶16 | ??1∶32 | ??1∶64 | ??1∶32 |
| Yolk antibody | ????1∶8 | ??1∶16 | ??1∶32 | ??1∶16 | ??1∶16 | ??1∶32 | ??1∶64 | ??1∶32 | |
| ??2 | Serum antibody | ????1∶8 | ??1∶16 | ??1∶64 | ??1∶8 | ??1∶16 | ??1∶8 | ??1∶16 | ??1∶32 |
| Yolk antibody | ????1∶8 | ??1∶16 | ??1∶32 | ??1∶8 | ??1∶16 | ??1∶32 | ??1∶64 | ??1∶32 | |
| ??3 | Serum antibody | ????0 | ??0 | ??1∶4 | ??1∶4 | ??1∶16 | ??1∶8 | ??1∶64 | ??1∶32 |
| Yolk antibody | ????0 | ??1∶8 | ??1∶16 | ??1∶16 | ??1∶16 | ??1∶16 | ??1∶16 | ??1∶32 | |
| ??4 | Serum antibody | ????1∶8 | ??1∶16 | ??1∶64 | ??1∶32 | ??1∶16 | ??1∶32 | ??1∶64 | ??1∶64 |
| Yolk antibody | ????1∶4 | ??1∶32 | ??1∶64 | ??1∶32 | ??1∶16 | ??1∶32 | ??1∶64 | ??1∶32 | |
| ??5 | Serum antibody | ????1∶8 | ??1∶16 | ??1∶32 | ??1∶32 | ??1∶8 | ??1∶16 | ??1∶64 | ??1∶32 |
| Yolk antibody | ????1∶16 | ??1∶32 | ??1∶16 | ??1∶8 | ??1∶16 | ??1∶32 | ??1∶64 | ??1∶32 | |
(1) serum antibody and yolk antibody are tired and are changed consistently, raise gradually with tiring of immunogen booster immunization antibody, and common three exempt from the back peaked in one week, reduced gradually then, four exempted to make antibody titer to reach higher level at this moment;
(2) two kinds of antibody titers are only variant in section sometime, but both total variation tendencies do not have significant difference;
(3) immunity back serum antibody titer rises earlier, and it is also very fast to descend to the certain level; Yolk antibody is tired and is changed slowlyer, but often keeps for some time after reaching certain level, and it is slower to descend;
(4) from every chicken antibody change curve, antibody titer there are differences between different immune bodies: antibody rises very soon after some individual immunity, and some then needs the long period just can reach higher antibody horizontal.
Conclusion: measure serum antibody titer and can react the change dynamics that yolk antibody is tired; For obtaining the yolk antibody that height is tired, at least should immunity three times, increase immune time and can make the height yolk antibody of tiring keep the long period.2.6 the purification of yolk antibody
Adopt polyoxyethylene glycol (PEG-6000) precipitator method.Get yolk liquid, add 0.01MPBS (pH7.4) the dilution yolk of 4 times of volumes, add the 3.5%PEG-6000 of 5 times of volumes then, fully dissolving back 5000rpm
*20min is centrifugal.Collect supernatant liquor, add 8.5%PEG-6000, fully dissolving back 5000rpm
*25min is centrifugal.Abandon supernatant liquor, precipitation is dissolved among the 0.01MPBS of 2.5 times of volumes, adds 12%PEG-6000, fully dissolving back 5000rpm
*25min is centrifugal.Precipitation is dissolved among the 0.01MPBS of 1/4 volume, is cooled to 0 ℃, adds 50% ethanol of 1/4 volume precooling, 10000rpm behind the mixing
*25min is centrifugal.Precipitation is dissolved among the 1/4 volume 0.01MPBS, dialyses 24 hours in the PBS of 50 times of volumes in 4 ℃, collects the back and adds 0.1% sodium azide, is stored in 4 ℃.2.7 the yolk antibody freeze-drying of purifying
The yolk antibody liquid of purifying is sub-packed in the cillin bottle in a small amount, and freeze-drying is spent the night on small-sized Freeze Drying Equipment, takes out and is stored in 4 ℃.Temperature is minimum in the freeze-drying process reduces to-38 ℃, keeps 3-4 hour to freeze-drying fully, returns to room temperature then gradually.
Embodiment 3 TGEV yolk antibody therapeutic tests
Get 1 age in days and do not eat just 12 of suckling piglet, the milk of feeding is observed and is divided into 4 groups after 1 day at random, and be respectively: attack malicious treatment group (4), attack poison and do not treat group (4), poison group (2) is not attacked in treatment, does not treat and does not attack malicious organize (2).The TGEV that to attack malicious used virus be ultrafiltration is directly oral after 10 times of dilutions of 0.01MPBS, treating used yolk antibody is the purification sample, it is 1: 64 that agar diffusion test mensuration yolk antibody is tired, same oral feeding judged the result of treatment of TGEV yolk antibody according to each group clinical symptom and mortality ratio.
Test-results shows: it is normal neither to attack poison group piglet growth, symptom of diarrhea do not occur, and whether the yolk antibody of feeding is to not influence of its growth; And two attack poison group piglet and serious symptom of diarrhea all occurs, show as yellow watery diarrhea, piglet becomes thin rapidly without all dead in the yolk antibody treatment group piglet three days, yolk antibody treatment group grice diarrhoea symptom then obviously alleviates, ight soil becomes soft excrement by watery diarrhea, and be shaped gradually, to recover gradually after the week, death condition does not appear.
Claims (5)
1. transmissible gastro-enteritis virus yolk antibody, it is characterized in that this antibody is that transmissible gastro-enteritis virus is used for immune non-immunization laying hen as artificial antigen, collect this by the egg that chicken produces of immunity, the transmissible gastro-enteritis virus yolk antibody that from its yolk, extracts.
2. the preparation method of the described transmissible gastro-enteritis virus yolk antibody of claim 1, it is characterized in that with the non-immunization laying hen of transmissible gastro-enteritis virus immunity, collect this by the egg that chicken produces of immunity, from its yolk, extract and purifying transmissible gastro-enteritis virus yolk antibody.
3. the preparation method of the described transmissible gastro-enteritis virus yolk antibody of claim 2, described from yolk, extract and purifying transmissible gastro-enteritis virus yolk antibody is to exempt from the back chicken two to begin to collect the egg that produces one week, isolate yolk, mix back homogenate with equivalent PBS, be prepared into yolk liquid, measure antibody titer, use polyoxyethylene glycol (PEG-6000) precipitator method to purify then.
4. the used artificial antigen of the preparation described transmissible gastro-enteritis virus yolk antibody of claim 1 is characterized in that this antigen is that transmissible gastro-enteritis virus is bred the cell toxicant that the back obtains on the pig testis cell.
5. the preparation method of the described artificial antigen of claim 4 the steps include: to catch and kill after 48 hours to the oral TGEV virus of piglet liquid, takes out small intestine, therefrom extracts and purifying TGEV, breeds on cell then.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02103667 CN1206243C (en) | 2002-02-08 | 2002-02-08 | Yolk antibody and antigen of pig's infective enterogastritis virus and its preparing process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02103667 CN1206243C (en) | 2002-02-08 | 2002-02-08 | Yolk antibody and antigen of pig's infective enterogastritis virus and its preparing process |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1382736A true CN1382736A (en) | 2002-12-04 |
| CN1206243C CN1206243C (en) | 2005-06-15 |
Family
ID=4739855
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 02103667 Expired - Fee Related CN1206243C (en) | 2002-02-08 | 2002-02-08 | Yolk antibody and antigen of pig's infective enterogastritis virus and its preparing process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1206243C (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1318449C (en) * | 2003-05-13 | 2007-05-30 | 雅臣药业集团（远东）有限公司 | Anti SARS specificity IgY and combination preparation thereof |
| CN102580088A (en) * | 2012-03-26 | 2012-07-18 | 广东紫金正天药业有限公司 | Egg yolk antibody for treating porcine circovirus disease and method for producing compound thereof |
| CN102908620A (en) * | 2011-08-04 | 2013-02-06 | 广州格拉姆生物科技有限公司 | Preparation method of egg yolk antibody injection for treating piglet diarrhea |
| CN102914647A (en) * | 2012-05-29 | 2013-02-06 | 安徽农业大学 | Method for detecting duck flavivirus antibody by using agar diffusion method |
| CN103777010A (en) * | 2014-01-27 | 2014-05-07 | 深圳市宝舜泰生物医药股份有限公司 | ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit for porcine epidemic diarrhea viruses and preparation method thereof |
| CN104788561A (en) * | 2014-01-17 | 2015-07-22 | 北京大北农科技集团股份有限公司动物医学研究中心 | Anti-swine transmissible gastroenteritis virus and porcine epidemic diarrhea virus egg-yolk antibody and preparation method thereof |
| CN106496325A (en) * | 2016-09-27 | 2017-03-15 | 中国农业科学院兰州畜牧与兽药研究所 | A kind of microcapsule is coated the preparation method and application of anti-main pathogen of bovine mastitis yolk antibody IgY |
| CN107880120A (en) * | 2017-11-06 | 2018-04-06 | 陕西诺威利华生物科技有限公司 | A kind of transmissible gastro-enteritis virus Yolk antibody and preparation method thereof |
| CN108130316A (en) * | 2018-02-09 | 2018-06-08 | 天康生物股份有限公司 | Porcine epidemic diarrhea virus and its separation method and application |
| CN108517013A (en) * | 2018-04-09 | 2018-09-11 | 国家食品安全风险评估中心 | Preparation method, zearalenone enzyme linked immunological kit and its detection method of zearalenone antibody |
| CN108676780A (en) * | 2018-07-24 | 2018-10-19 | 华中农业大学 | A method of efficiently separating pig enteric coronavirus virus |
| CN113121679A (en) * | 2021-04-01 | 2021-07-16 | 仲恺农业工程学院 | High-immunity yolk antibody for pigeon Newcastle disease and preparation method and application thereof |
-
2002
- 2002-02-08 CN CN 02103667 patent/CN1206243C/en not_active Expired - Fee Related
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1318449C (en) * | 2003-05-13 | 2007-05-30 | 雅臣药业集团（远东）有限公司 | Anti SARS specificity IgY and combination preparation thereof |
| CN102908620A (en) * | 2011-08-04 | 2013-02-06 | 广州格拉姆生物科技有限公司 | Preparation method of egg yolk antibody injection for treating piglet diarrhea |
| CN102580088A (en) * | 2012-03-26 | 2012-07-18 | 广东紫金正天药业有限公司 | Egg yolk antibody for treating porcine circovirus disease and method for producing compound thereof |
| CN102914647A (en) * | 2012-05-29 | 2013-02-06 | 安徽农业大学 | Method for detecting duck flavivirus antibody by using agar diffusion method |
| CN104788561B (en) * | 2014-01-17 | 2019-07-16 | 北京大北农科技集团股份有限公司动物医学研究中心 | Anti-swine infectious enterogastritis virus and Porcine epidemic diarrhea virus Yolk antibody and preparation method thereof |
| CN104788561A (en) * | 2014-01-17 | 2015-07-22 | 北京大北农科技集团股份有限公司动物医学研究中心 | Anti-swine transmissible gastroenteritis virus and porcine epidemic diarrhea virus egg-yolk antibody and preparation method thereof |
| CN103777010A (en) * | 2014-01-27 | 2014-05-07 | 深圳市宝舜泰生物医药股份有限公司 | ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit for porcine epidemic diarrhea viruses and preparation method thereof |
| CN103777010B (en) * | 2014-01-27 | 2016-04-20 | 深圳市宝舜泰生物医药股份有限公司 | porcine epidemic diarrhea virus ELISA detection kit and preparation method thereof |
| CN106496325A (en) * | 2016-09-27 | 2017-03-15 | 中国农业科学院兰州畜牧与兽药研究所 | A kind of microcapsule is coated the preparation method and application of anti-main pathogen of bovine mastitis yolk antibody IgY |
| CN107880120A (en) * | 2017-11-06 | 2018-04-06 | 陕西诺威利华生物科技有限公司 | A kind of transmissible gastro-enteritis virus Yolk antibody and preparation method thereof |
| CN107880120B (en) * | 2017-11-06 | 2018-10-12 | 陕西诺威利华生物科技有限公司 | A kind of transmissible gastro-enteritis virus Yolk antibody and preparation method thereof |
| CN108130316A (en) * | 2018-02-09 | 2018-06-08 | 天康生物股份有限公司 | Porcine epidemic diarrhea virus and its separation method and application |
| CN108517013A (en) * | 2018-04-09 | 2018-09-11 | 国家食品安全风险评估中心 | Preparation method, zearalenone enzyme linked immunological kit and its detection method of zearalenone antibody |
| CN108676780A (en) * | 2018-07-24 | 2018-10-19 | 华中农业大学 | A method of efficiently separating pig enteric coronavirus virus |
| CN108676780B (en) * | 2018-07-24 | 2021-08-10 | 华中农业大学 | Method for efficiently separating porcine enterocoronavirus |
| CN113121679A (en) * | 2021-04-01 | 2021-07-16 | 仲恺农业工程学院 | High-immunity yolk antibody for pigeon Newcastle disease and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1206243C (en) | 2005-06-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP2548115B2 (en) | Passive immunization of mammals with avian antibodies | |
| CN101144062B (en) | Lactobacillus casei strain and application for products thereof in bird immunity | |
| CN1206243C (en) | Yolk antibody and antigen of pig's infective enterogastritis virus and its preparing process | |
| AU2010319047A1 (en) | Therapeutic agent for psoriasis or atopic dermatitis | |
| CN106344917A (en) | Swine fever mucosal immunity live vaccine composition and preparation method of vaccine | |
| US20020044942A1 (en) | Transfer factor composition and process for producing same | |
| CN107177001A (en) | It is a kind of to prevent and treat Yolk antibody of Porcine Epidemic Diarrhea and preparation method thereof | |
| CN115120711A (en) | Recombinant protein vaccine for haemaphysalis qinghai and preparation method thereof | |
| CN108969492B (en) | Oral attenuated freeze-dried vaccine for swine fever and preparation method thereof | |
| CN114539396A (en) | Bovine enterovirus egg yolk antibody and preparation | |
| CN100412092C (en) | Bacillus coli resisting chicken yolk antibody, its preparation and use | |
| CN104888213A (en) | Preparation method of classical swine fever spleen-lymph-sourced compound living vaccine | |
| CN108295053B (en) | Application of the schizandrin in enhancing PEDV vaccine immune response | |
| CN1509765A (en) | Freezed dry preparation of lecithal antibody and its producing method | |
| KR101518473B1 (en) | Immunogloblin y complex composition for vaccinating or curing canine viral infectious diseases and production method thereof | |
| CN102488899A (en) | Preparation method and application of yolk immunoglobulin sustained-release preparation | |
| CN112358545B (en) | Preparation method and application of pigeon trichomonas and candida albicans bigeminal egg yolk antibody powder | |
| KR100267746B1 (en) | Oral immunological preparation containing egg-yolk antibodies, for prevention and treatment of porcine diarrhea caused by enterotoxigenic escherichia coli | |
| CN108703952B (en) | Freeze-drying protective agent for swine fever oral attenuated freeze-dried vaccine and application | |
| CN103446182A (en) | Preparation method of specific transfer factor for highly pathogemc pathogenic porcine reproductive and respiratory syndrome | |
| CN1384119A (en) | Composite yolk antibody for resisting fowl's viral blight and its prepn and application | |
| CN112625126A (en) | anti-Hafnia alvei yolk antibody and preparation method and application thereof | |
| CN112079916B (en) | Canine parvovirus egg yolk antibody and preparation method thereof | |
| RU2145876C1 (en) | Method of stimulating immune response against animal parasitic helminths | |
| CN111467488B (en) | Water-soluble composite immunoadjuvant and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20050615 Termination date: 20140208 |