CN1092979C - Method for preparing polyclonal antibody by using waterfowl - Google Patents
Method for preparing polyclonal antibody by using waterfowl Download PDFInfo
- Publication number
- CN1092979C CN1092979C CN99109616A CN99109616A CN1092979C CN 1092979 C CN1092979 C CN 1092979C CN 99109616 A CN99109616 A CN 99109616A CN 99109616 A CN99109616 A CN 99109616A CN 1092979 C CN1092979 C CN 1092979C
- Authority
- CN
- China
- Prior art keywords
- polyclonal antibody
- antibody
- aquatic bird
- antigen
- duck
- 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.)
- Expired - Lifetime
Links
- 238000000034 method Methods 0.000 title claims abstract description 25
- 241000272517 Anseriformes Species 0.000 title abstract description 7
- 239000000427 antigen Substances 0.000 claims abstract description 14
- 102000036639 antigens Human genes 0.000 claims abstract description 14
- 108091007433 antigens Proteins 0.000 claims abstract description 14
- 241000272525 Anas platyrhynchos Species 0.000 claims abstract description 12
- 238000006243 chemical reaction Methods 0.000 claims abstract description 9
- 241000124008 Mammalia Species 0.000 claims abstract description 7
- 108060003951 Immunoglobulin Proteins 0.000 claims abstract description 5
- 102000018358 immunoglobulin Human genes 0.000 claims abstract description 5
- 235000013601 eggs Nutrition 0.000 claims description 12
- 102000004169 proteins and genes Human genes 0.000 claims description 9
- 108090000623 proteins and genes Proteins 0.000 claims description 9
- 238000000746 purification Methods 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 4
- 210000003205 muscle Anatomy 0.000 claims description 3
- 241000272814 Anser sp. Species 0.000 claims description 2
- 241000700605 Viruses Species 0.000 claims description 2
- 230000000844 anti-bacterial effect Effects 0.000 claims description 2
- 206010013663 drug dependence Diseases 0.000 claims description 2
- 238000010353 genetic engineering Methods 0.000 claims description 2
- 229940088597 hormone Drugs 0.000 claims description 2
- 239000005556 hormone Substances 0.000 claims description 2
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 2
- 238000007920 subcutaneous administration Methods 0.000 claims description 2
- 208000011117 substance-related disease Diseases 0.000 claims description 2
- 239000000084 colloidal system Substances 0.000 abstract description 10
- 238000010790 dilution Methods 0.000 abstract description 10
- 239000012895 dilution Substances 0.000 abstract description 10
- 210000002969 egg yolk Anatomy 0.000 abstract description 10
- 125000001165 hydrophobic group Chemical group 0.000 abstract description 6
- 238000004587 chromatography analysis Methods 0.000 abstract description 5
- 238000001962 electrophoresis Methods 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- 238000011033 desalting Methods 0.000 abstract description 2
- 238000000502 dialysis Methods 0.000 abstract description 2
- 238000003452 antibody preparation method Methods 0.000 abstract 1
- 239000007788 liquid Substances 0.000 description 12
- 102000002322 Egg Proteins Human genes 0.000 description 9
- 108010000912 Egg Proteins Proteins 0.000 description 9
- 235000002639 sodium chloride Nutrition 0.000 description 9
- 150000003839 salts Chemical class 0.000 description 8
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 7
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 7
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 7
- 235000011130 ammonium sulphate Nutrition 0.000 description 7
- 229960003010 sodium sulfate Drugs 0.000 description 7
- 229910052938 sodium sulfate Inorganic materials 0.000 description 7
- 235000011152 sodium sulphate Nutrition 0.000 description 7
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- 238000010828 elution Methods 0.000 description 6
- 238000001556 precipitation Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 235000013345 egg yolk Nutrition 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 229920000936 Agarose Polymers 0.000 description 4
- 241000272522 Anas Species 0.000 description 4
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 239000012141 concentrate Substances 0.000 description 4
- 210000004681 ovum Anatomy 0.000 description 4
- 241000894007 species Species 0.000 description 4
- GUPXYSSGJWIURR-UHFFFAOYSA-N 3-octoxypropane-1,2-diol Chemical compound CCCCCCCCOCC(O)CO GUPXYSSGJWIURR-UHFFFAOYSA-N 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 229920002684 Sepharose Polymers 0.000 description 3
- 239000002671 adjuvant Substances 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 230000008485 antagonism Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 238000003018 immunoassay Methods 0.000 description 3
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 208000023275 Autoimmune disease Diseases 0.000 description 2
- 241000233855 Orchidaceae Species 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- SOGAXMICEFXMKE-UHFFFAOYSA-N alpha-Methyl-n-butyl acrylate Natural products CCCCOC(=O)C(C)=C SOGAXMICEFXMKE-UHFFFAOYSA-N 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000003599 detergent Substances 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 230000036737 immune function Effects 0.000 description 2
- 238000000760 immunoelectrophoresis Methods 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- LNGIIXGLYTUUHJ-UHFFFAOYSA-N 2-methylbut-2-enoic acid methyl 2-methylprop-2-enoate Chemical compound COC(=O)C(C)=C.CC=C(C)C(O)=O LNGIIXGLYTUUHJ-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 239000012619 Butyl Sepharose® Substances 0.000 description 1
- 241000272834 Cairina moschata Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 102000004895 Lipoproteins Human genes 0.000 description 1
- 108090001030 Lipoproteins Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- VVQNEPGJFQJSBK-UHFFFAOYSA-N Methyl methacrylate Chemical compound COC(=O)C(C)=C VVQNEPGJFQJSBK-UHFFFAOYSA-N 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- UXFAMLIXCYVDDW-UHFFFAOYSA-N butyl 2-methylprop-2-enoate 2,4,4-trimethylpent-2-enoic acid Chemical compound C(C)(C)(C)C=C(C(=O)O)C.C(CCC)OC(C(=C)C)=O UXFAMLIXCYVDDW-UHFFFAOYSA-N 0.000 description 1
- 238000013098 chemical test method Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000010612 desalination reaction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000000385 dialysis solution Substances 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000003147 molecular marker Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 238000013094 purity test Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000008399 tap water Substances 0.000 description 1
- 235000020679 tap water Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention relates to a method for preparing polyclonal antibody by waterfowl, which comprises collecting the egg of laying duck after the egg duck is injected with antigen to induce antibody, taking out the yolk, purifying the yolk, carrying out hydrophobic group colloid chromatography, dialysis, desalting, concentration and SDS-acrylamide colloid electrophoresis, and analyzing to obtain yolk immunoglobulin with purity over 96% and high purity and without Fc end. The results of the analysis show that the antibody preparation method not only can prepare polyclonal antibodies with high specificity and dilution capability, but also can effectively avoid the false positive reaction generated by the polyclonal antibodies prepared by taking mammals as intermediate hosts.
Description
The present invention relates to a kind of method of utilizing immunoreation to prepare antibody, particularly relate to the method for utilizing palmipeds to prepare polyclonal antibody for intermediate host.
Polyclonal antibody (polyclonal antibldy) is a most important parts in the immunological test reagent.Be widely used in every biomedical sector now.The preparation of polyclonal antibody in the past all utilizes mammal (as white mice, rabbit, cattle, sheep etc.) and makes.Yet, the event of biological species poor (Phylogenetic distance), usually need a large amount of antigens can produce ideal antibody, but with the made immunity inspection reagent of the polyclonal antibody of this kind method gained when the clinical practice, the puzzlement of generation false positive reaction (false positive) is arranged more.The generation main cause of this reaction is that the antibody (IgG in the animal serum) of gained is all mammal with its source of a corpse or other object for laboratory examination and chemical testing (IgG in the human serum), have due to the biologically so-called homogeneity (homogenecity), especially when human body has autoimmune disease (autoimmune disease), such as the rheumatoid factor (Rheumatoid factor) being arranged when existing, the easier interference that causes this kind false positive reaction.Be immune intermediate host abroad in recent years, in it is laid eggs, make polyclonal antibody with laying hen (flying bird).Discovery is far away because of the biological species difference, can reduce to the interference of false positive reaction minimum in clinical practice.And if when being antigen with the human body protein also because of the biological species difference is far away, and immunity is preferably arranged.Because Taiwan is quite flourishing aspect industries such as foster duck and every product processing thereof, it is intermediate host that the present invention mainly utilizes laying ducks (palmipeds) to replace laying hen, produces polyclonal antibody to reach the purpose of extensive use.
Purpose of the present invention is to provide a kind of and utilizes aquatic bird to prepare the method for polyclonal antibody for intermediate host, and this antibody has high specific (specificity) and dilution capacity, more can effectively avoid false positive reaction.This kind method is with the advantage that with the laying hen is the intermediate host comparison: 1, the immunoreation of laying hen only reaches 1/3 of laying ducks; 2, the weight of Ovum Anas domestica is to two times of egg, and this means can produce one to duple polyclonal antibody; 3, the mortality rate of laying ducks is low far beyond laying hen, and this is a special character of the present invention.
The present invention relates to a kind of method of utilizing aquatic bird to prepare polyclonal antibody, it comprises:
(a) antigen (immunogen) with purification is injected to aquatic bird;
(b) strengthen reaction with Freund again;
(c) after 5-15 days, promptly there is polyclonal antibody to produce in the laying eggs of aquatic bird.
Utilize in the method that aquatic bird prepares polyclonal antibody of the present invention, wherein said injection comprises methods such as muscle, subcutaneous, Intradermal, intravascular injection; Described aquatic bird is selected from duck, female goose; Described antigen comprises human immunoglobulin;
Utilize in the method that aquatic bird prepares polyclonal antibody of the present invention, wherein said antigen is selected from:
(a) from protein, the hormone of mammal;
(b) various viruses;
(c) antibacterial;
(d) recombiant protein for preparing with genetic engineering, synthetic peptide (syntheticpeptide);
The protein conjugate of (e) various medicines for treatment, drug dependence medicine.
The present invention relates to a kind of polyclonal antibody, it is characterized in that, it is to utilize to comprise that the method for above-mentioned steps (a)-(c) is prepared.
The invention still further relates to the purposes of described polyclonal antibody, it is applied to various clinical immunologic function test reagents and research is used in the preparation and food and feed of immunoreagent.
The invention further relates to a kind of in aquatic bird is laid eggs the method for purification polyclonal antibody, it comprises:
(1) with water is solvent, crude separation IgY in the egg yolk that this is laid eggs, can use tap water, purification distilled water, deionized water, or for little acid of PH 〉=5.1 or contain nonionic detergent (non-ionic detergent) aqueous solution or 0.15-1.0M common salt aqueous solution below 0.05%, phosphate buffer or TRIS buffer (Tris-buffer) etc.;
(2) add ammonium sulfate, make the saturation that reaches 30-50%, or add sodium sulfate, make the concentration that reaches 14-20% (W/V), under its concentration, repeat to saltout;
(3) use and to contain hydrophobic group colloid (hydrophobic ligand coupled tomatrices) and carry out chromatography IgY:
(a) described hydrophobic group colloid is selected from:
Butyl-agarose (Butyl sepharose), octyl sepharose (Octylsepharose), phenyl sepharose (Phenyl sepharose), alkyl agarose (Alkylsepharose), methyl methacrylate (Methyl methacrylate) and methacrylic acid butyl ester (T-Butyl methacrylate);
(b) operating condition of chromatography tubing string:
(i) the tubing string level pad is at PH7.0-8.0;
(ii) level pad is ammonium sulfate or the sodium sulfate of 1.0M, and its conductivity is 110-200ms/ml;
(iii) type of elution can be with gradient elution or single liquid eluting:
Gradient elution: A liquid be aforementioned b (ii),
B liquid is for the buffer of salt,
Single liquid eluting: in advance A liquid and B liquid being mixed and adjusts its conductivity is 45-80ms/ml.
In aquatic bird is laid eggs in the method for purification polyclonal antibody, wherein said laying eggs laid eggs or through the storage egg of 4 ℃ of preservations for fresh of the present invention.
Brief description of drawings:, can have more fully the present invention and understand via following description and diagram.Wherein:
Fig. 1: utilizing 10%SDS-acryloyl electrophoresis (SDS-PAGE) to cooperate Kumasi orchid (commassie blue, trade name) is dyestuff, decision antibody molecule amount size and purity assay.Molecular weight unit kda is 1000 dalton (dalton) among the figure.
1.-9. meaning is as follows in Fig. 1:
1. salt precipitation, 100 times of dilutions, not reducing conditions.
2. salt precipitation, 10 times of dilutions, reducing condition.
3. salt precipitation, 10 times of dilutions, not reducing conditions.
4. colloid chromatography, secondary peak top gleanings, reducing condition.
5. colloid chromatography, secondary peak top gleanings, not reducing condition.
6. concentrate finished product, reducing condition.
7. concentrate finished product, not reducing condition.
8. concentrate finished product, reducing condition.
9. standard molecular weight reagent (Molecular Marker), 29～200kda
Fig. 2: enzyme immunoassay (EIA) is measured the antibody dilution ability.
In Fig. 2 the meaning of vertical coordinate OD450 for the optical density that under the 450nm wavelength, records (optical density, OD).
Fig. 3: agar immunoelectrophoresis figure.This measures the specificity and the purity of antibody.
The meaning of each labelling is as follows in Fig. 3:
Ab
1The IgY antibody of → duck antagonism human IgG (full molecule)
Ab
2The IgY antibody of → duck antagonism human IgG (full molecule)
Ab
3The antibody of → rabbit antagonism duck IgG (W)
Ag
1→ human IgG (w)
Ag
2→ human albumin
Ag
3→ human whole serum
Ag
4→ rabbit whole blood is clear
Ag
5→ it is Ab
1
Ag
6→ human IgG (w)
Following embodiment is used to further specify the present invention, but these embodiment only are not in order to restriction the present invention as preferred example.
Embodiment 1: the preparation of polyclonal antibody
Laying ducks is a palmipeds, and the difference of highly significant is arranged with mammals on the biological species difference, so when being antigen with the protein in mammals source, easily obtaining good immunoreation (Immuneresponse) and obtain better antibody.Getting the human immunoglobulin (human IgG) that healthy laying ducks (as dish duck, Beijing duck, Muscovy duck and change duck etc.) finishes with purification in advance is antigen with Freund's complete adjuvant (Complete Freund ' s adjuvant) with 1: 1 mixed, with the muscle injection mode injection, generate with induce antibody.Afterwards, strengthen reaction (booster dose) with Freund (IncompleteFreund ' s adjuvant) with 1: 1 ratio again.About ten day after tomorrow, promptly there is antibody to generate in the Ovum Anas domestica.
Embodiment 2: the antibody list is from reaching purification
Egg yolk accounts for duck 1/3 weight, and its main component is a lipoprotein, accounts for 85%.Contained Yolk immune globulin (Yolk Immunologlobulin in the egg yolk; IgY) belonging to water solublity, therefore can utilize this characteristic, is solvent with water, and IgY is single from coming out in egg yolk.Get one piece in Ovum Anas domestica, this Ovum Anas domestica is fresh lay eggs or through the storage egg of 4 ℃ of preservations, break to take out the water that egg yolk adds the about 6-20 of its volume times, utilizes general agitator or magnetic stirrer (magnetic stirrer) with its mix homogeneously.Then with high-speed refrigerated centrifuge under the state of 4 ℃ and 5000rpm, separated, and collected the supernatant that is rich in IgY.
Embodiment 3: saltout
Add ammonium sulfate (Ammonium sulfate (NH
4)
2SO
4) or sodium sulfate (sodiumsulfate Na
2SO
4) in the supernatant that is rich in IgY, make to reach 30-50% ammonium sulfate saturation, or the concentration of 14-20% (w/v) sodium sulfate, to saltout.After full and uniform stirring, utilize high-speed refrigerated centrifuge equally under the state of 4 ℃ and 5000rpm, separated, and collected its precipitation.Generally speaking, repeat to saltout and centrifugal two steps, can effectively improve product purity.The protein of different molecular weight has different binding abilities with the colloid of band hydrophobic group (hydrophobic ligand) under the environment of variable concentrations.The hydrophobic group colloid is selected from following material: butyl-agarose, octyl sepharose, phenyl sepharose, alkyl agarose, methyl methacrylate and methacrylic acid butyl ester.
By fill the tubing string (column) form with the hydrophobic group colloid, then with band salt (as ammonium sulfate or sodium sulfate etc.) and be not with two kinds of buffer of salt with the alternately washing of gradient mode, type of elution can be with gradient elution or single liquid eluting with the salt precipitation thing.Promptly with ammonium sulfate or the sodium sulfate of 1.0M to 1.5M, its conductivity is 110-200ms/ml to gradient elution A liquid, and B liquid is the buffer of not being with salt.And single liquid eluting promptly in advance A liquid and B liquid to be mixed and adjusts its conductivity be 45-80ms/ml.Collect the secondary peak of cleaning mixture, with phosphate buffer (Phosphate buffersaline) dialysis desalting.After treating that desalination is finished, dialysis solution is condensed into includes the 1.0mg/ml protein solution to concentrate film (ultrafiltrationmembrane), this is the antibody behind the purification.
Embodiment 4: purity testing and antibody ability are measured
The molecular weight of IgY is about 170,000 (170kDa), and removing its molecular weight of Fc end back is 110,000 (110kDa).Electrophoretic techniques is at present in order to the method for the most normal use of decision molecular weight size and purity assay aspect.Therefore the present invention utilizes SDS-acrylamide electrophoresis (SDS-PAGE) to cooperate the Kumasi orchid to be dyestuff, with various antibody samples colloid dyeing sentence read result such as Fig. 1 after electrophoretic analysis.The sample of the 9th row is a standard molecular weight 29-200kDa reagent among Fig. 1.
Embodiment 5: the mensuration of antibody dilution ability and antibody specificity
Utilize enzyme immunoassay (EIA) (enzyme immuno assay; EIA) dilution capacity of analysis antibody, experimental result such as Fig. 2.Symbolic representation among Fig. 2 is at the different resulting antibody of time the method according to this invention.
Embodiment 6: the agar directional diffusion is analyzed (gel double diffusion)
(3a) antibody dilution ability: the antibody dilution ability that records during for antigen with the human IgG is more than or equal to 32 times.
(3b) antibody specificity: with the agar immunoelectrophoresis analysis, experimental result such as Fig. 3.With rabbit igg, when goat IgG and Mus IgG are antigen, all do not have precipitation line and occur, this shows that this kind polyclonal antibody is splendid to the specificity of IgG.
Embodiment 7: the purposes of polyclonal antibody
The method according to this invention (embodiment 1), utilize the antigen of different structure, can prepare and have corresponding specific polyclonal antibody, these antibody just can be used for clinical immunologic function test reagent, research immunoreagent, or add in food, the feedstuff and make the eater have immunity.
Claims (5)
1. method of utilizing aquatic bird to prepare polyclonal antibody, it comprises:
(a) antigen with purification is injected to aquatic bird;
(b) strengthen reaction with Freund again;
(c) after 5-15 days, promptly there is polyclonal antibody to produce in the laying eggs of aquatic bird.
2. the method for utilizing aquatic bird to prepare polyclonal antibody as claimed in claim 1, wherein said injection comprise muscle, subcutaneous, Intradermal, intravascular injection method.
3. the method for utilizing aquatic bird to prepare polyclonal antibody as claimed in claim 1, wherein said aquatic bird is selected from duck, female goose.
4. the method for utilizing aquatic bird to prepare polyclonal antibody as claimed in claim 1, wherein said antigen comprises human immunoglobulin.
5. as claim 1 or the 4 described methods of utilizing aquatic bird to prepare polyclonal antibody, wherein said antigen is selected from:
(a) from protein, the hormone of mammal;
(b) various viruses;
(c) antibacterial;
(d) recombiant protein for preparing with genetic engineering, synthetic peptide;
The protein conjugate of (e) each medicine for treatment, drug dependence medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99109616A CN1092979C (en) | 1999-07-01 | 1999-07-01 | Method for preparing polyclonal antibody by using waterfowl |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN99109616A CN1092979C (en) | 1999-07-01 | 1999-07-01 | Method for preparing polyclonal antibody by using waterfowl |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 02106193 Division CN1439650A (en) | 2002-04-08 | 2002-04-08 | Method for preparing polyclonal antibody by using waterfowl |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1280833A CN1280833A (en) | 2001-01-24 |
| CN1092979C true CN1092979C (en) | 2002-10-23 |
Family
ID=5274022
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN99109616A Expired - Lifetime CN1092979C (en) | 1999-07-01 | 1999-07-01 | Method for preparing polyclonal antibody by using waterfowl |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1092979C (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP5300200B2 (en) * | 2006-08-31 | 2013-09-25 | 大阪府 | Rapid diagnostic method specific to avian influenza virus |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1081882A (en) * | 1993-04-15 | 1994-02-16 | 山东省齐河县兽药研究所 | Antibacterial agent for poultry and compound method |
| CN1093927A (en) * | 1993-04-22 | 1994-10-26 | 上海医科大学 | A kind of novel immunogenic complex formulation and preparation method |
-
1999
- 1999-07-01 CN CN99109616A patent/CN1092979C/en not_active Expired - Lifetime
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1081882A (en) * | 1993-04-15 | 1994-02-16 | 山东省齐河县兽药研究所 | Antibacterial agent for poultry and compound method |
| CN1093927A (en) * | 1993-04-22 | 1994-10-26 | 上海医科大学 | A kind of novel immunogenic complex formulation and preparation method |
Non-Patent Citations (3)
| Title |
|---|
| 中国预防兽医学报1999,21(1) 1999.1.31 田国斌鸭源禽流感病毒的分离和鉴定 * |
| 扬州大学学报1998,(1) 1998.1.31 王宝安等鸭乙肝病毒表面抗原特异性单抗孤研制及应用 * |
| 扬州大学学报1998,(1) 1998.1.31 王宝安等鸭乙肝病毒表面抗原特异性单抗孤研制及应用;中国预防兽医学报1999,21(1) 1999.1.31 田国斌鸭源禽流感病毒的分离和鉴定 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1280833A (en) | 2001-01-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5420253A (en) | Method for purifying egg yolk immunoglobulins | |
| Cooper | General immunology | |
| US20140031528A1 (en) | Method for selecting plasma cells or plasmablasts, method for producing target antigen specific antibodies, and novel monoclonal antibodies | |
| CA1215919A (en) | Detection of malignant tumor cells | |
| CN1811436A (en) | Method for detecting Ennoxacin and its special enzyme-linked immune reagent kit | |
| CN1233828C (en) | Antihuman hemoglobin detection reagent strips and monoclone antibody contained therewith | |
| CN103215230A (en) | Hybridoma cell strain AFM1B7, monoclonal antibody thereof and aflatoxin M1 flow lag immunization time-resolved fluorescence quick test kit | |
| CN1414017A (en) | Preparation of anti cytoplasmic thymidine kinase-IgY and tumor diagnosis composition | |
| CN1159580C (en) | Reagent for colloidal gold chromatographic analysis of SARS coronavirus antigen | |
| CN1092979C (en) | Method for preparing polyclonal antibody by using waterfowl | |
| CN1296567A (en) | Method and apparatus for detecting conception in animals | |
| CN1439650A (en) | Method for preparing polyclonal antibody by using waterfowl | |
| CN101880316A (en) | Human RBPMS polypeptide and preparation method of antibody thereof | |
| CN116284370B (en) | Multimeric glycosylated hemoglobin monoclonal antibody and preparation method thereof | |
| CN1858064A (en) | Chicken yoke antibody resisting Bengalese cobra venom and its preparation | |
| CN1237346C (en) | Immunoassay of human medullarin and diagnosis of multiple sclerosis using the same | |
| AU2018203565B2 (en) | Reticulocyte growth factor, and preparation method and application thereof | |
| TW471970B (en) | Method of production of polyclonal antibody by duck | |
| CA2171317C (en) | Method for purifying egg yolk immunoglobulins | |
| CN1811453A (en) | Animal source food animal medicine residue detection reagent kit and detecting method thereof | |
| CN1294418C (en) | Method and reagent box for inspecting mycelian protein antibody of white candida | |
| CN1453589A (en) | Reagent for colloidal gold chromatographic analysis of SARS coronavirus antibody | |
| CN110655575B (en) | Epitope polyclonal antibody of toxoplasma gondii Actin protein and application thereof | |
| CN86104525A (en) | The analytical approach of human interleukin-2 and reagent | |
| UA144483U (en) | METHOD OF STRENGTHENING REACTION OF PRECIPITATION BY IMMUNOELECTROPHORESIS |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20021023 |